Updated on 2021/12/08

写真a

 
SHIBATA, Shigenobu
 
Affiliation
Faculty of Science and Engineering, School of Advanced Science and Engineering
Job title
Professor
Profile
九州大学、薬学部、薬学研究科
時間生物学、時間栄養学、時間運動学

Concurrent Post

  • Affiliated organization   Global Education Center

  • Faculty of Commerce   School of Commerce

  • Faculty of Science and Engineering   Graduate School of Advanced Science and Engineering

Research Institute

  • 2020
    -
    2022

    理工学術院総合研究所   兼任研究員

  • 2019
    -
    2021

    時間栄養学研究所   プロジェクト研究所所長

Education

  •  
    -
    1956

    Kyushu University   Graduate School, Division of Pharmaceutical Sciences  

  •  
    -
    1951

    Kyushu University   Faculty of Pharmaceutical Science  

Degree

  • (BLANK)

  • 九州大学   薬学博士

Professional Memberships

  •  
     
     

    Society for Research on Biological Rhythms

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    日本神経科学会

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    Society research on biological Rhythms

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    日本神経精神薬理学会

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    日本薬学会

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    日本時間生物学会

  •  
     
     

    日本薬理学会

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Research Areas

  • Family and consumer sciences, and culture and living

Research Interests

  • 健康科学

  • 時間生物学

  • 時間運動学

  • 時間栄養学

Papers

  • Changes in sleep phase and body weight of mobile health App users during COVID-19 mild lockdown in Japan.

    Yu Tahara, Takae Shinto, Kosuke Inoue, Farnaz Roshanmehr, Akito Ito, Mikiko Michie, Shigenobu Shibata

    International journal of obesity (2005)   45 ( 10 ) 2277 - 2280  2021.10  [International journal]

     View Summary

    OBJECTIVE AND METHOD: The stay-at-home order during the COVID-19 pandemic has restricted individuals' social behaviors, and therefore, effected their lifestyle including sleep, diet, and physical activity. Using the cross-sectional study design with a large sample size (N = 30,275) from the mobile health App users in Japan, we show age-dependent lifestyle changes during a nonpunitive "mild lockdown" (from April to May 2020). RESULTS: Sleep onset and offset were delayed on work-days but not on free-days with increased sleep duration and decreased social jetlag, and the changes were more evident in the younger population. Although average weight change was close to none because of the users' characteristic (95% of App users try to lose weight), we investigated an association between lifestyle change and body-weight change. Participants who reported advanced sleep phase during mild lockdown described a weight decrease. In contrast, the delayed sleep phase reported a weight gain. The results were significant after adjustment of confounding factors including physical activity and meal changes. CONCLUSIONS: Although there is cumulative evidence showing a relationship between late chronotype and obesity, it is still unclear about the potential benefit of the chronotype management to control body weight. Thus, to the best of our knowledge, this is the first study investigating the association between chronotype and weight changes by leveraging a large cohort.

    DOI PubMed

  • Distribution of dietary protein intake in daily meals influences skeletal muscle hypertrophy via the muscle clock

    Shinya Aoyama, Hyeon-Ki Kim, Rina Hirooka, Mizuho Tanaka, Takeru Shimoda, Hanako Chijiki, Shuichi Kojima, Keisuke Sasaki, Kengo Takahashi, Saneyuki Makino, Miku Takizawa, Masaki Takahashi, Yu Tahara, Shigeki Shimba, Kazuyuki Shinohara, Shigenobu Shibata

    Cell Reports   36 ( 1 ) 109336 - 109336  2021.07

    DOI

  • Inhomogeneous and Anisotropic Mechanical Properties of the Triceps Surae Aponeuroses in Older Adults: Relationships With Muscle Strength and Walking Performance.

    Xiyao Shan, Pavlos Evangelidis, Takaki Yamagishi, Shun Otsuka, Fumiko Tanaka, Shigenobu Shibata, Yasuo Kawakami

    Journal of aging and physical activity     1 - 9  2021.01  [International journal]

     View Summary

    This study investigated (a) site- and direction-dependent variations of passive triceps surae aponeurosis stiffness and (b) the relationships between aponeurosis stiffness and muscle strength and walking performance in older individuals. Seventy-nine healthy older adults participated in this study. Shear wave velocities of the triceps surae aponeuroses at different sites and in two orthogonal directions were obtained in a prone position at rest using supersonic shear imaging. The maximal voluntary isometric contraction torque of the plantar flexors and normal (preferred) and fast (fastest possible) walking speeds (5-m distance) were also measured. The shear wave velocities of the adjoining aponeuroses were weakly associated with plantar flexion torque (r = .23-.34), normal (r = .26), and fast walking speed (r = .25). The results show clear spatial variations and anisotropy of the triceps surae aponeuroses stiffness in vivo, and the aponeurosis stiffness was associated with physical ability in older adults.

    DOI PubMed

  • Use of a social jetlag-mimicking mouse model to determine the effects of a two-day delayed light- and/or feeding-shift on central and peripheral clock rhythms plus cognitive functioning.

    Atsushi Haraguchi, Yutaro Nishimura, Miyabi Fukuzawa, Yosuke Kikuchi, Yu Tahara, Shigenobu Shibata

    Chronobiology international     1 - 17  2020.12  [International journal]

     View Summary

    Social jetlag (SJL) is defined as the discrepancy between social and biological rhythms and calculated by the difference between the midpoint of sleep time on working-days and free-days. Previous human and mouse studies showed SJL is positively related to evening chronotype and significantly related to smoking habit, cardiovascular risk, cognitive ability, and that SJL-mimicking conditions, simulating the real lifestyle situation of SJL in many humans, disrupt the regularity of estrous cycles of female animals. The effects of SJL-mimicking conditions on circadian rhythms and cognitive function and the reasons why the discrepancy between social and biological rhythms is involved in SJL have not yet been investigated. Therefore, in this study, we utilized a mouse model of SJL-mimicking conditions - 6-hour delayed-light/dark (LD) conditions for 2 days and normal-LD conditions for the following 5 days - applied for several weeks during which biological rhythms were monitored. Circadian rhythms of central and peripheral clocks and metabolism of the mice under the SJL-mimicking condition were always delayed for 2-3 hours compared with those under the normal-LD condition. Moreover, SJL-mimicking conditions impaired their cognitive function using a novel object recognition test. Only the delayed timing of either the light phase of the LD or of feeding for 2 days, comparable to the free-days situation of humans, delayed the circadian staging of rhythms the following 5 days. Furthermore, sleep deprivation during the early mornings for 5 days, which is comparable to early rise times experienced by humans on working-days and does affect the staging of circadian rhythms (circadian misalignment schedule), delayed the locomotor activity rhythms the next 2 days, comparable to free-days in humans, which is similar to the lifestyle rhythm of the evening chronotype. Our results demonstrated that the circadian misalignment schedule for 5 days changed the locomotor activity rhythms the following 2 days to the evening chronotype, that light- and/or feeding-shift conditions for 2 days exacerbate SJL, and that SJL-mimicking conditions delay the metabolic rhythm and cause cognitive impairment.

    DOI PubMed

  • Effect of the Intake of a Snack Containing Dietary Fiber on Postprandial Glucose Levels

    Hyeon-Ki Kim, Takuya Nanba, Mamiho Ozaki, Hanako Chijiki, Masaki Takahashi, Mayuko Fukazawa, Jin Okubo, Shigenobu Shibata

    Foods   9 ( 10 ) 1500 - 1500  2020.10

     View Summary

    To examine the effects of the intake of a snack containing dietary fiber under free-living conditions on postprandial glucose levels in older adults, nine healthy older adults aged 76.9 ± 1.6 years (mean ± standard error) completed two crossover trials: 1) regular snack (BISCUIT) intake and 2) intake of snacks with a high dietary fiber content (DF-BISCUIT). In both trials, each participant consumed either BISCUIT or DF-BISCUIT between lunch and dinner time for 1 week. During the intervention, the blood glucose levels of all the subjects were observed using a continuous glucose monitoring system. Lower 24 h blood glucose levels were yielded in the DF-BISCUIT than the BISCUIT trials. Moreover, compared to the BISCUIT trials, the blood glucose levels after dinner and areas under the curve (AUCs) were significantly decreased in the DF-BISCUIT treatments. The blood glucose levels and AUCs after the intake of the next day’s breakfast were suppressed in the DF-BISCUIT treatments compared to those in the BISCUIT trials. Our data indicate that the intake of snacks with a high dietary fiber content under free-living conditions is an effective way to restrain postprandial glucose levels and that the effect lasts until breakfast the next day.

    DOI

  • Screen Time Duration and Timing: Effects on Obesity, Physical Activity, Dry Eyes, and Learning Ability in Elementary School Children

    Yui Mineshita, Hyeon-Ki Kim, Hanako Chijiki, Takuya Nanba, Takae Shinto, Shota Furuhashi, Satoshi Oneda, Mai Kuwahara, Anzu Suwama, Shigenobu Shibata

       2020.10

     View Summary

    <title>Abstract</title>
    <bold>Background: </bold>As internet use becomes more widespread, the screen time (ST) of elementary school students increases yearly. It is known that longer durations of ST can affect obesity, physical activity, dry eye disease, and learning ability. However, the effects of ST just before bedtime have not been clarified. Therefore, we examined ST duration and timing effects on elementary school children. <bold>Methods: </bold>We conducted a survey of 7,419 elementary school students in Tokyo, Japan using a questionnaire on food education. ST duration and timing (just before bedtime) served as the explanatory variables, and the relationship between obesity, physical activity, dry eyes, and learning ability was analyzed using logistic regression analysis. Gender, school year, height, and weight were considered confounding factors. First, we examined whether ST duration and timing were related to each objective variable, using a univariate model to examine all variables. Thereafter, we performed multivariate logistic regression analyses for all variables showing a significant difference in the univariate models.<bold>Results:</bold> The results showed that the relationship between the ST duration and obesity, physical activity, and learning ability was statistically significant. The relationship between ST timing and obesity, dry eyes, and learning ability was also statistically significant. Therefore, ST timing has a greater effect on dry eyes, and ST duration has a greater effect on academic performance. <bold>Conclusion: </bold>Our findings indicate that ST in school children is related to obesity, physical activity, dry eyes, and learning ability, and they suggest that not only the duration but also the timing of ST is important.

    DOI

  • Ingestion of Helianthus tuberosus at Breakfast Rather Than at Dinner Is More Effective for Suppressing Glucose Levels and Improving the Intestinal Microbiota in Older Adults.

    Hyeon-Ki Kim, Hanako Chijiki, Takuya Nanba, Mamiho Ozaki, Hiroyuki Sasaki, Masaki Takahashi, Shigenobu Shibata

    Nutrients   12 ( 10 )  2020.10  [International journal]

     View Summary

    To date, nutritional studies have focused on the total intake of dietary fiber rather than intake timing. In this study, we examined the effect of the timing of daily Helianthustuberosus ingestion on postprandial and 24 h glucose levels, as well as on intestinal microbiota in older adults. In total, 37 healthy older adults (age = 74.9 ± 0.8 years) were recruited. The participants were randomly assigned to either a morning group (MG, n = 18) or an evening group (EG, n = 17). The MG and EG groups were instructed to take Helianthustuberosus powder (5 g/day) just before breakfast or dinner, respectively, for 1 week after the 1-week control period. The glucose levels of all participants were monitored using a continuous glucose monitoring system throughout the 2 weeks. The intestinal microbiota was analyzed by sequencing 16S rRNA genes from feces before and after the intervention. There were no significant differences in the physical characteristics or energy intake between groups. Helianthustuberosus intake led to decreases in tissue glucose levels throughout the day in both groups (p < 0.01, respectively). As a result of examining the fluctuations in tissue glucose levels up to 4 hours after each meal, significant decreases in the areas under the curves (AUCs) were observed for all three meals after intervention, but only in the MG (breakfast: p = 0.012, lunch: p = 0.002, dinner: p = 0.005). On the other hand, in the EG, there was a strong decrease in the AUC after dinner, but only slight decreases after breakfast and lunch (breakfast: p = 0.017, lunch: p = 0.427, dinner: p = 0.002). Moreover, the rate of change in the peak tissue glucose level at breakfast was significantly decreased in the MG compared to the EG (p = 0.027). A greater decrease was observed in the change in the blood glucose level after the ingestion of Helianthustuberosus in the MG than in the EG. Furthermore, the relative abundance of Ruminococcus in the MG at the genus level was significantly higher at baseline than in the EG (p = 0.016) and it was also significantly lower after the intervention (p = 0.013). Our findings indicate that Helianthustuberosus intake in the morning might have relatively stronger effects on the intestinal microbiota and suppress postprandial glucose levels to a greater extent than when taken in the evening.

    DOI PubMed

  • 時間栄養学

    金 鉉基, 柴田 重信

    体力科学   69 ( 5 ) 401 - 411  2020.10

  • Gamma oryzanol impairs alcohol-induced anxiety-like behavior in mice via upregulation of central monoamines associated with Bdnf and Il-1β signaling.

    Salina Akter, Kazi Rasel Uddin, Hiroyuki Sasaki, Yijin Lyu, Shigenobu Shibata

    Scientific reports   10 ( 1 ) 10677 - 10677  2020.06  [International journal]

     View Summary

    Adolescent alcohol exposure may increase anxiety-like behaviors by altering central monoaminergic functions and other important neuronal pathways. The present study was designed to investigate the anxiolytic effect of 0.5% γ-oryzanol (GORZ) and its neurochemical and molecular mechanisms under chronic 10% ethanol consumption. Five-week-old ICR male mice received either control (14% casein, AIN 93 M) or GORZ (14% casein, AIN 93 M + 0.5% GORZ) diets in this study. We showed that GORZ could potentially attenuate alcohol-induced anxiety-like behaviors by significantly improving the main behavioral parameters measured by the elevated plus maze test. Moreover, GORZ treatment significantly restored the alcohol-induced downregulation of 5-hydroxytryptophan and 5-hydroxyindole acetic acid in the hippocampus and improved homovanillic acid levels in the cerebral cortex. Furthermore, a recovery increase in the level of 3-methoxy-4-hydroxyphenylglycol both in the hippocampus and cerebral cortex supported the anxiolytic effect of GORZ. The significant elevation and reduction in the hippocampus of relative mRNA levels of brain-derived neurotrophic factor and interleukin 1β, respectively, also showed the neuroprotective role of GORZ in ethanol-induced anxiety. Altogether, these results suggest that 0.5% GORZ is a promising neuroprotective drug candidate with potential anxiolytic, neurogenic, and anti-neuroinflammatory properties for treating adolescent alcohol exposure.

    DOI PubMed

  • Combinatorial Effects of Soluble, Insoluble, and Organic Extracts from Jerusalem Artichokes on Gut Microbiota in Mice.

    Hiroyuki Sasaki, Yijin Lyu, Yuki Nakayama, Fumiaki Nakamura, Aya Watanabe, Hiroki Miyakawa, Yoichi Nakao, Shigenobu Shibata

    Microorganisms   8 ( 6 )  2020.06  [International journal]

     View Summary

    Jerusalem artichokes contain high amounts of inulin, which is a prebiotic that supports digestive health, as well as a variety of insoluble fibers and caffeoylquinic acid. The individual impact of these components on gut microbiota is well known; however, the combinatorial effects are less clear. In this investigation, we fractionated Jerusalem artichokes into three parts (water-soluble extract, insoluble extract, and organic extract) and powdered them. Mice were fed a high-fat diet that included one or more of these extracts for 10 days, and then their cecal pH, cecal short-chain fatty acids (SCFAs), and fecal microbiota were evaluated. The combination of the water-soluble and organic extract decreased cecal pH and increased the concentration of SCFAs and led to dynamic changes in the composition of the gut microbiota. These results demonstrate that both the water-soluble and organic extracts in Jerusalem artichokes are bioactive substances that are capable of changing SCFA production and the composition of gut microbiota. Powdered Jerusalem artichokes, rather than inulin supplements, may be superior for promoting a healthy gut.

    DOI PubMed

  • Consumption of Biscuits with a Beverage of Mulberry or Barley Leaves in the Afternoon Prevents Dinner-Induced High, but Not Low, Increases in Blood Glucose among Young Adults

    Mai Kuwahara, Hyeon-Ki Kim, Mamiho Ozaki, Takuya Nanba, Hanako Chijiki, Mayuko Fukazawa, Jin Okubo, Yui Mineshita, Masaki Takahashi, Shigenobu Shibata

    Nutrients   12 ( 6 ) 1580 - 1580  2020.05

     View Summary

    We examined the impact of consuming biscuits with a beverage of powdered mulberry or barley leaves in the afternoon on postprandial glucose levels at dinnertime among young adults. A total of 18 young adults participated in a partially double-blinded, randomized crossover trial over 2 weeks, consuming either: (1) no biscuits; (2) a biscuit; (3) a biscuit with a beverage of powdered mulberry leaves; or (4) a biscuit with a beverage of powdered barley leaves, as an afternoon snack followed by a standardized test dinner. Glucose levels were recorded after each meal. Results showed intake of biscuits with a beverage of mulberry and barley leaves significantly reduced postprandial rises in glucose after their immediate consumption and dinner, though there was no direct relationship between the glucose levels at the two meals. Compared to those with low glucose levels, participants with high glucose levels at dinner showed a stronger second meal effect, that was attributed to the mulberry or barley leaves, and were also more likely to have lean body weights and prefer evenings. Our findings indicate that eating snacks alongside mulberry or barley leaves is an effective way to suppress postprandial glucose levels in young adults with high glucose levels who prefer evenings.

    DOI

  • 1日におけるタンパク質の摂取配分は骨格筋の肥大に影響する

    青山 晋也, 廣岡 里菜, 高橋 健吾, 田原 優, 篠原 一之, 柴田 重信

    日本生理学雑誌   82 ( 2 ) 34 - 34  2020.05

  • 1日におけるタンパク質の摂取配分は骨格筋の肥大に影響する

    青山 晋也, 廣岡 里菜, 高橋 健吾, 田原 優, 篠原 一之, 柴田 重信

    日本生理学雑誌   82 ( 2 ) 34 - 34  2020.05  [Refereed]

  • ウロリチンAのPer2遺伝子発現リズムに対するインビトロ評価

    杜 堯, 折原 芳波, 原口 敦嗣, 立石 法史, 柴田 重信

    日本栄養・食糧学会大会講演要旨集   74回   233 - 233  2020.04

  • Effects of Timing of Acute and Consecutive Catechin Ingestion on Postprandial Glucose Metabolism in Mice and Humans.

    Masaki Takahashi, Mamiho Ozaki, Miku Tsubosaka, Hyeon-Ki Kim, Hiroyuki Sasaki, Yuji Matsui, Masanobu Hibi, Noriko Osaki, Masashi Miyashita, Shigenobu Shibata

    Nutrients   12 ( 2 )  2020.02  [Refereed]  [International journal]

     View Summary

    We examined the effects of the timing of acute and consecutive epigallocatechin gallate (EGCG) and catechin-rich green tea ingestion on postprandial glucose in mice and human adults. In mouse experiments, we compared the effects of EGCG administration early (morning) and late (evening) in the active period on postprandial glucose. In human experiments, participants were randomly assigned to the morning-placebo (MP, n = 10), morning-green tea (MGT, n = 10), evening-placebo (EP, n = 9), and evening-green tea (EGT, n = 9) groups, and consumed either catechin-rich green tea or a placebo beverage for 1 week. At baseline and after 1 week, participants consumed their designated beverages with breakfast (MP and MGT) or supper (EP and EGT). Venous blood samples were collected in the fasted state and 30, 60, 120, and 180 min after each meal. Consecutive administration of EGCG in the evening, but not in the morning, reduced postprandial glucose at 30 (p = 0.006) and 60 (p = 0.037) min in the evening trials in mice. In humans, ingestion of catechin-rich green tea in the evening decreased postprandial glucose (three-factor analysis of variance, p < 0.05). Thus, catechin intake in the evening more effectively suppressed elevation of postprandial glucose.

    DOI PubMed

  • Circadian rhythm and its association with birth and infant outcomes: research protocol of a prospective cohort study.

    Satvinder Kaur, Ai Ni Teoh, Nurul Husna Mohd Shukri, Siti Raihanah Shafie, Normina Ahmad Bustami, Masaki Takahashi, Pei Jean Lim, Shigenobu Shibata

    BMC pregnancy and childbirth   20 ( 1 ) 96 - 96  2020.02  [Refereed]  [International journal]

     View Summary

    BACKGROUND: Circadian rhythm plays an important role as our internal body's clock that synchronizes behavior and physiology according to the external 24-h light-dark cycle. Past studies have associated disrupted circadian rhythm with higher risk of miscarriages, preterm birth and low birth weights. This paper described the protocol of a prospective cohort study which aims to determine the circadian rhythm in pregnant women, identify its association with maternal factors during pregnancy, gestational weight gain, birth and infant outcomes. METHODS: Ten government maternal and child health clinics in Kuala Lumpur, Malaysia will be randomly selected. Sample size of 438 first-trimester pregnant women will be followed-up until the birth of their infant. Salivary melatonin and cortisol concentration among subsample will be determined using enzyme-linked immunosorbent assay. Data on sleep quality, psychological distress and morningness/eveningness chronotype of pregnant women will be collected using validated questionnaires. Pedometer will be used to measure 5-day physical activity data. Total gestational weight gain will be determined at the end of pregnancy. Utilization of 3-day food record is to capture meal timing and nutrient intake. All measurements will be done in 2nd and 3rd trimester. Birth outcomes will be collected through clinic records and Centers for Disease Control and Prevention (CDC) Neonatal questionnaire. Infants will be followed-up at 6 and 12 months old to obtain anthropometric measurements. DISCUSSION: There is a growing recognition of the role of maternal circadian rhythm, which entrains fetal circadian rhythms that may subsequently have long-term health consequences. The present study will identify the effect of circadian rhythm on pregnancy outcomes and infant growth in the first year of life.

    DOI PubMed

  • Effect of Dose and Timing of Burdock (Arctium lappa) Root Intake on Intestinal Microbiota of Mice.

    Aya Watanabe, Hiroyuki Sasaki, Hiroki Miyakawa, Yuki Nakayama, Yijin Lyu, Shigenobu Shibata

    Microorganisms   8 ( 2 )  2020.02  [International journal]

     View Summary

    Water-soluble dietary fiber such as inulin improves the beta diversity of the intestinal microbiota of mice fed with a high-fat diet (HFD). The circadian clock is the system that regulates the internal daily rhythm, and it affects the pattern of beta diversity in mouse intestinal microbiota. Burdock (Arctium lappa) root contains a high concentration of inulin/fructan (approximately 50%) and is a very popular vegetable in Japan. Arctium lappa also contains functional substances that may affect intestinal microbiota, such as polyphenols. We compared the effects of inulin and A. lappa powder on the diversity of the intestinal microbiota of HFD-fed mice. 16S rDNA from the intestinal microbiota obtained from feces was analyzed by 16S Metagenomic Sequencing Library Preparation. It was found to have a stronger effect on microbiota than inulin alone, suggesting that inulin has an additive and/or synergic action with other molecules in A. lappa root. We examined the effects of intake timing (breakfast or dinner) of A. lappa on intestinal microbiota. The intake of A. lappa root in the evening had a stronger effect on microbiota diversity in comparison to morning intake. Therefore, it is suggested that habitual consumption of A. lappa root in the evening may aid the maintenance of healthy intestinal microbiota.

    DOI PubMed

  • The circadian clock is disrupted in mice with adenine-induced tubulointerstitial nephropathy.

    Hiroaki Motohashi, Yu Tahara, Daniel S Whittaker, Huei-Bin Wang, Takahiro Yamaji, Hiromichi Wakui, Atsushi Haraguchi, Mayu Yamazaki, Hiroki Miyakawa, Koki Hama, Hiroyuki Sasaki, Tomoko Sakai, Rina Hirooka, Kengo Takahashi, Miku Takizawa, Saneyuki Makino, Shinya Aoyama, Christopher S Colwell, Shigenobu Shibata

    Kidney international    2020.01  [Refereed]  [International journal]

     View Summary

    Chronic Kidney Disease (CKD) is increasing in incidence and has become a worldwide health problem. Sleep disorders are prevalent in patients with CKD raising the possibility that these patients have a disorganized circadian timing system. Here, we examined the effect of adenine-induced tubulointerstitial nephropathy on the circadian system in mice. Compared to controls, adenine-treated mice showed serum biochemistry evidence of CKD as well as increased kidney expression of inflammation and fibrosis markers. Mice with CKD exhibited fragmented sleep behavior and locomotor activity, with lower degrees of cage activity compared to mice without CKD. On a molecular level, mice with CKD exhibited low amplitude rhythms in their central circadian clock as measured by bioluminescence in slices of the suprachiasmatic nucleus of PERIOD 2::LUCIFERASE mice. Whole animal imaging indicated that adenine treated mice also exhibited dampened oscillations in intact kidney, liver, and submandibular gland. Consistently, dampened circadian oscillations were observed in several circadian clock genes and clock-controlled genes in the kidney of the mice with CKD. Finally, mice with a genetically disrupted circadian clock (Clock mutants) were treated with adenine and compared to wild type control mice. The treatment evoked worse kidney damage as indicated by higher deposition of gelatinases (matrix metalloproteinase-2 and 9) and adenine metabolites in the kidney. Adenine also caused non-dipping hypertension and lower heart rate. Thus, our data indicate that central and peripheral circadian clocks are disrupted in the adenine-treated mice, and suggest that the disruption of the circadian clock accelerates CKD progression.

    DOI PubMed

  • Chrono-nutrition

    Kim Hyeon-Ki, Shibata Shigenobu

    Japanese Journal of Physical Fitness and Sports Medicine   69 ( 5 ) 401 - 411  2020

     View Summary

    <p>In mammals, the circadian rhythms have been shown to regulate several physiological functions, including body temperature, sleep-wake behavior, physical activity, hormonal secretions, and metabolism. These processes are controlled by circadian clock genes, and abnormal circadian rhythms are associated with the development of obesity, diabetes, and lifestyle-related diseases. In addition, the timing of behaviors such as food intake, exercise, and stress influence circadian rhythms, including clock gene expression in peripheral tissues. Therefore, the interaction between nutrition and the circadian clock is so-called "chrono-nutrition" is poised to become an important research field of chronobiology. In this review, we review the effects of a timed-nutrition on circadian clocks and their timing-dependent effects on physiological functions.</p>

    DOI CiNii

  • Gamma Oryzanol Alleviates High-Fat Diet-Induced Anxiety-Like Behaviors Through Downregulation of Dopamine and Inflammation in the Amygdala of Mice.

    Salina Akter, Kazi Rasel Uddin, Hiroyuki Sasaki, Shigenobu Shibata

    Frontiers in pharmacology   11   330 - 330  2020  [International journal]

     View Summary

    Background: A high-fat diet (HFD) can induce obesity and metabolic disorders that are closely associated with cognitive impairments, and the progression of several psychiatric disorders such as anxiety. We have previously demonstrated the anxiolytic-like effect of Gamma oryzanol (GORZ) in chronic restraint stressed mice. Objective: We studied the neurochemical and molecular mechanisms that underlie the preventive effect of GORZ in HFD-induced anxiety-like behaviors, monoaminergic dysfunction, and inflammation. Methods: Eight-week-old Institute of Cancer (ICR) male mice weighing 33-34 g were divided into the following groups and free-fed for 8 weeks: control (14% casein, AIN 93M); HFD; HFD + GORZ (0.5% GORZ). Body weight gain was checked weekly. The anxiolytic-like effects of GORZ were examined via open-field test (OFT) and elevated plus maze (EPM) test. Brain levels of monoamines [5-hydroxy tryptamine (5-HT), dopamine (DA), and norepinephrine (NE)] and their metabolites [5-hydroxyindole acetic acid (5-HIAA), homovanillic acid (HVA), and 3-methoxy-4-hydroxyphenylglycol (MHPG)], proinflammatory cytokines such as tumor necrosis factor-αα (Tnf-α) mRNA levels, and interleukin 1-β (Il-1β) mRNA levels in the cerebral cortex and amygdala were examined using high-performance liquid chromatography-electrochemical detection (HPLC-ECD), and real-time reverse transcription-polymerase chain reaction (RT-PCR), respectively. Results: Mice fed a HFD for eight weeks showed anxiety-like behaviors in association with HFD-induced body weight gain. GORZ potentially blocked HFD-induced anxiety-like behaviors via significant improvement of the primary behavioral parameters in behavioral tests, with a minor reduction in HFD-induced body weight gain. Furthermore, GORZ treatment significantly downregulated HFD-induced upregulation of dopamine levels in the brain's amygdala. Significant reduction of the relative mRNA expression of Tnf-α and Il-1 β was also observed in the amygdala of HFD + GORZ mice, compared to HFD mice. Conclusions: Our findings strongly suggest that 0.5% GORZ exerts anxiolytic-like effects, possibly through downregulation of dopamine, and via expression of proinflammatory cytokines Tnf-α and Il-1 β in the case of chronic HFD exposure.

    DOI PubMed

  • Time-of-Day-Dependent Physiological Responses to Meal and Exercise.

    Shinya Aoyama, Shigenobu Shibata

    Frontiers in nutrition   7   18 - 18  2020  [Refereed]  [International journal]

     View Summary

    The mammalian circadian clock drives the temporal coordination in cellular homeostasis and it leads the day-night fluctuation of physiological functions, such as sleep/wake cycle, hormonal secretion, and body temperature. The mammalian circadian clock system in the body is classified hierarchically into two classes, the central clock in the suprachiasmatic nucleus (SCN) of the hypothalamus and the peripheral clocks in peripheral tissues such as the intestine and liver, as well as other brain areas outside the SCN. The circadian rhythm of various tissue-specific functions is mainly controlled by each peripheral clock and partially by the central clock as well. The digestive, absorptive, and metabolic capacities of nutrients also show the day-night variations in several peripheral tissues such as small intestine and liver. It is therefore indicated that the bioavailability or metabolic capacity of nutrients depends on the time of day. In fact, the postprandial response of blood triacylglycerol to a specific diet and glucose tolerance exhibit clear time-of-day effects. Meal frequency and distribution within a day are highly related to metabolic functions, and optimal time-restricted feeding has the potential to prevent several metabolic dysfunctions. In this review, we summarize the time-of-day-dependent postprandial response of macronutrients to each meal and the involvement of circadian clock system in the time-of-day effect. Furthermore, the chronic beneficial and adverse effects of meal time and eating pattern on metabolism and its related diseases are discussed. Finally, we discuss the timing-dependent effects of exercise on the day-night variation of exercise performance and therapeutic potential of time-controlled-exercise for promoting general health.

    DOI PubMed

  • Effect of different sources of dietary protein on muscle hypertrophy in functionally overloaded mice.

    Shinya Aoyama, Rina Hirooka, Takeru Shimoda, Shigenobu Shibata

    Biochemistry and biophysics reports   20   100686 - 100686  2019.12  [Refereed]  [International journal]

     View Summary

    Dietary protein intake is important for skeletal muscle protein synthesis. In this study, we investigated the differential effect of protein sources on hypertrophy of plantaris muscle induced by surgical ablation of gastrocnemius and soleus muscles. Six-week old mice were fed diets containing caseinate, whey, or soy as protein sources for 2 weeks. Plantaris muscle hypertrophy was induced by a unilateral ablation of synergistic muscles after a week. Food intake of soy protein-fed mice was higher than that of caseinate and whey-fed mice, resulting in higher body and fat weights. Plantaris muscle weight in sham-operated mice was not different across the groups. Overload-operated plantaris muscle weight and increased ratio of overloaded muscle to sham-operated muscle weights were higher in caseinate-fed mice than in whey- and soy protein-fed mice, suggesting caseinate as a promising protein source for muscle hypertrophy.

    DOI PubMed

  • Administration timing and duration-dependent effects of sesamin isomers on lipid metabolism in rats.

    Tateishi N, Morita S, Yamazaki I, Okumura H, Kominami M, Akazawa S, Funaki A, Tomimori N, Rogi T, Shibata H, Shibata S

    Chronobiology international     1 - 17  2019.12  [Refereed]

    DOI PubMed

  • The Timing Effects of Soy Protein Intake on Mice Gut Microbiota.

    Tamura K, Sasaki H, Shiga K, Miyakawa H, Shibata S

    Nutrients   12 ( 1 )  2019.12  [Refereed]  [International journal]

     View Summary

    Soy protein intake is known to cause microbiota changes. While there are some reports about the effect of soy protein intake on gut microbiota and lipid metabolism, effective timing of soy protein intake has not been investigated. In this study, we examined the effect of soy protein intake timing on microbiota. Mice were fed twice a day, in the morning and evening, to compare the effect of soy protein intake in the morning with that in the evening. Mice were divided into three groups: mice fed only casein protein, mice fed soy protein in the morning, and mice fed soy protein in the evening under high-fat diet conditions. They were kept under the experimental condition for two weeks and were sacrificed afterward. We measured cecal pH and collected cecal contents and feces. Short-chain fatty acids (SCFAs) from cecal contents were measured by gas chromatography. The microbiota was analyzed by sequencing 16S rRNA genes from feces. Soy protein intake whether in the morning or evening led to a greater microbiota diversity and a decrease in cecal pH resulting from SCFA production compared to casein intake. In addition, these effects were relatively stronger by morning soy protein intake. Therefore, soy protein intake in the morning may have relatively stronger effects on microbiota than that in the evening.

    DOI PubMed

  • Eurotium Cristatum Fermented Okara as a Potential Food Ingredient to Combat Diabetes.

    Li Yan Chan, Masaki Takahashi, Pei Jean Lim, Shinya Aoyama, Saneyuki Makino, Ferdinandus Ferdinandus, Shi Ya Clara Ng, Satoshi Arai, Hideaki Fujita, Hong Chang Tan, Shigenobu Shibata, Chi-Lik Ken Lee

    Scientific reports   9 ( 1 ) 17536 - 17536  2019.11  [Refereed]  [International journal]

     View Summary

    Type 2 diabetes mellitus (T2DM) is a chronic disease, and dietary modification is a crucial part of disease management. Okara is a sustainable source of fibre-rich food. Most of the valorization research on okara focused more on the physical attributes instead of the possible health attributes. The fermentation of okara using microbes originated from food source, such as tea, sake, sufu and yoghurt, were explored here. The aim of this study is to investigate fermented okara as a functional food ingredient to reduce blood glucose levels. Fermented and non-fermented okara extracts were analyzed using the metabolomic approach with UHPLC-QTof-MSE. Statistical analysis demonstrated that the anthraquinones, emodin and physcion, served as potential markers and differentiated Eurotium cristatum fermented okara (ECO) over other choices of microbes. The in-vitro α-glucosidase activity assays and in-vivo mice studies showed that ECO can reduce postprandial blood glucose levels. A 20% ECO loading crispy snack prototype revealed a good nutrition composition and could serve as a fundamental formulation for future antidiabetes recipe development, strengthening the hypothesis that ECO can be used as a novel food ingredient for diabetic management.

    DOI PubMed

  • Effects of timing of acute catechin-rich green tea ingestion on postprandial glucose metabolism in healthy men.

    Takahashi M, Ozaki M, Miyashita M, Fukazawa M, Nakaoka T, Wakisaka T, Matsui Y, Hibi M, Osaki N, Shibata S

    The Journal of nutritional biochemistry   73   108221  2019.11  [Refereed]

    DOI PubMed

  • Mice Microbiota Composition Changes by Inulin Feeding with a Long Fasting Period under a Two-Meals-Per-Day Schedule.

    Sasaki H, Miyakawa H, Watanabe A, Nakayama Y, Lyu Y, Hama K, Shibata S

    Nutrients   11 ( 11 )  2019.11  [Refereed]  [International journal]

     View Summary

    Water-soluble dietary fiber is known to modulate fecal microbiota. Although there are a few reports investigating the effects of fiber intake timing on metabolism, there are none on the effect of intake timing on microbiota. Therefore, in this study, we examined the timing effects of inulin-containing food on fecal microbiota. Mice were housed under conditions with a two-meals-per-day schedule, with a long fasting period in the morning and a short fasting period in the evening. Then, 10-14 days after inulin intake, cecal content and feces were collected, and cecal pH and short-chain fatty acids (SCFAs) were measured. The microbiome was determined using 16S rDNA sequencing. Inulin feeding in the morning rather than the evening decreased the cecal pH, increased SCFAs, and changed the microbiome composition. These data suggest that inulin is more easily digested by fecal microbiota during the active period than the inactive period. Furthermore, to confirm the effect of fasting length, mice were housed under a one-meal-per-day schedule. When the duration of fasting was equal, the difference between morning and evening nearly disappeared. Thus, our study demonstrates that consuming inulin at breakfast, which is generally after a longer fasting period, has a greater effect on the microbiota.

    DOI PubMed

  • Effects of day-time feeding on murine skeletal muscle growth and synthesis

    Shinya Aoyama, Shuichi Kojima, Keisuke Sasaki, Takeru Shimoda, Kengo Takahashi, Rina Hirooka, Yu Tahara, Shigenobu Shibata

    Journal of Nutrition and Intermediary Metabolism   17  2019.09  [Refereed]

    DOI

  • Correlation among clock gene expression rhythms, sleep quality, and meal conditions in delayed sleep-wake phase disorder and night eating syndrome.

    Haraguchi A, Komada Y, Inoue Y, Shibata S

    Chronobiology international   36 ( 6 ) 770 - 783  2019.06  [Refereed]

    DOI PubMed

  • Phase resetting of circadian peripheral clocks using human and rodent diets in mouse models of type 2 diabetes and chronic kidney disease.

    Yasuda S, Iwami S, Tamura K, Ikeda Y, Kamagata M, Sasaki H, Haraguchi A, Miyamatsu M, Hanashi S, Takato Y, Shibata S

    Chronobiology international   36 ( 6 ) 851 - 869  2019.06  [Refereed]  [International journal]

     View Summary

    The expression rhythms of clock genes, such as Per1, Per2, Bmal1, and Rev-erb α, in mouse peripheral clocks, are entrained by a scheduled feeding paradigm. In terms of food composition, a carbohydrate-containing diet is reported to cause strong entrainment through insulin secretion. However, it is unknown whether human diets entrain peripheral circadian clocks. In this study, we used freeze-dried diets for type 2 diabetes (DB) and chronic kidney disease (CKD), as well as low-carbohydrate diets. After 24 h of fasting, PER2::LUC knock-in mice were given access to food for 2 days during inactive periods, and bioluminescence rhythm was then measured using an in vivo imaging system. AIN-93M, the control mouse diet with a protein:fat:carbohydrate (PFC) ratio of 14.7:9.5:75.8, caused a significant phase advance (7.3 h) in the liver clock compared with that in 24 h fasted mice, whereas human diets caused significant but smaller phase advances (4.7-6.2 h). Compared with healthy and high fat/sucrose-induced DB mice, adenine-induced CKD mice showed attenuation of a phase-advance with a normal diet. There were no significant differences in phase-advance values between human diets (normal, DB, and CKD). In addition, a normal-carbohydrate diet (PFC ratio of 20.3:23.3:56.4) and a low-carbohydrate diet (PFC ratio of 36.4:42.9:20.7) caused similar phase advances in peripheral clocks. The present results strongly suggest that scheduled feeding with human diets can cause phase advances in the peripheral clocks of not only healthy, but also DB and CKD mice. This discovery provides support to the food-induced entrainment of peripheral clocks in human clinical trials.

    DOI PubMed

  • Effect of piceatannol on circadian Per2 expression in vitro and in vivo

    Yamamoto T, Iwami S, Aoyama S, Maruki-Uchida H, Mori S, Hirooka R, Takahashi K, Morita M, Shibata S

    Journal of Functional Foods   56   49 - 56  2019.05  [Refereed]

    DOI

  • Systemic oscillator-driven and nutrient-responsive hormonal regulation of daily expression rhythms for gluconeogenic enzyme genes in the mouse liver.

    Taira A, Arita E, Matsumoto E, Oohira A, Iwase K, Hiwasa T, Yokote K, Shibata S, Takiguchi M

    Chronobiology international   36 ( 5 ) 591 - 615  2019.05  [Refereed]  [International journal]

     View Summary

    Gluconeogenesis is de novo glucose synthesis from substrates such as amino acids and is vital when glucose is lacking in the diurnal nutritional fluctuation. Accordingly, genes for hepatic gluconeogenic enzymes exhibit daily expression rhythms, whose detailed regulations under nutritional variations remain elusive. As a first step, we performed general systematic characterization of daily expression profiles of gluconeogenic enzyme genes for phosphoenolpyruvate carboxykinase (PEPCK), cytosolic form (Pck1), glucose-6-phosphatase (G6Pase), catalytic subunit (G6pc), and tyrosine aminotransferase (TAT) (Tat) in the mouse liver. On a standard diet fed ad libitum, mRNA levels of these genes showed robust daily rhythms with a peak or an elevation phase during the late sleep-fasting period in the diurnal feeding/fasting (wake/sleep) cycle. The rhythmicity was preserved in constant darkness, modulated with prolonged fasting, attenuated by Clock mutation, and entrained to varied photoperiods and time-restricted feedings. These results are concordant with the notion that gluconeogenic enzyme genes are under the control of the intrinsic circadian oscillator, which is entrained by the light/dark cycle, and which in turn entrains the feeding/fasting cycle and also drives systemic signaling pathways such as the hypothalamic-pituitary-adrenal axis. On the other hand, time-restricted feedings also showed that the ingestion schedule, when separated from the light/dark cycle, can serve as an independent entrainer to daily expression rhythms of gluconeogenic enzyme genes. Moreover, nutritional changes dramatically modified expression profiles of the genes. In addition to prolonged fasting, a high-fat diet and a high-carbohydrate (no-protein) diet caused modification of daily expression rhythms of the genes, with characteristic changes in profiles of glucoregulatory hormones such as corticosterone, glucagon, and insulin, as well as their modulators including ghrelin, leptin, resistin, glucose-dependent insulinotropic polypeptide (GIP), and glucagon-like peptide-1 (GLP-1). Remarkably, high-protein (60% casein or soy-protein) diets activated the gluconeogenic enzyme genes atypically during the wake-feeding period, with paradoxical up-regulation of glucagon, which frequently formed correlation networks with other humoral factors. Based on these results, we propose that daily expression rhythms of gluconeogenic enzyme genes are under the control of systemic oscillator-driven and nutrient-responsive hormones.

    DOI PubMed

  • The effect of night shift work on the expression of clock genes in beard hair follicle cells.

    Miwa Hattammaru, Yu Tahara, Tomoko Kikuchi, Kiyotaka Okajima, Koichi Konishi, Shun Nakajima, Kyoko Sato, Kuniaki Otsuka, Hiroshi Sakura, Shigenobu Shibata, Takashi Nakaoka

    Sleep medicine   56   164 - 170  2019.04  [Refereed]  [International journal]

     View Summary

    OBJECTIVE: Shift work encompasses a broad range of work time arrangements. However, how shift work affects the circadian expression of clock genes remains to be explored. The objective of this study was to evaluate the pattern of clock gene expression in shift workers in the field. METHODS: We examined clock gene expression in Japanese men who work: (1) one night shift followed by a day off (caregivers: nurses and doctors; the one-night group); (2) three or more consecutive night shifts (factory workers; the consecutive-night group); or (3) daytime only (the daytime group), using beard follicle samples. The expression of Period3, Nuclear Receptor Subfamily 1 Group D Member 1 (Nr1d1), and Nuclear Receptor Subfamily 1 Group D Member 2 (Nr1d2) was examined by real-time polymerase chain reaction. RESULTS: Period3 expression in the daytime and one-night groups together with Nr1d2 expression in the one-night group fitted a 24-h-period cosine curve better than in the consecutive-night group (p = 0.004, 0.012, and 0.001, respectively). The level of overall Period3 gene expression, calibrated with that of 18S-rRNA, was decreased in the consecutive-night group compared with that in the daytime group (p = 0.006). The patterns of Period3 and Nr1d2 expression in the daytime and one-night groups were more coherent than those in the consecutive-night group. CONCLUSIONS: These results suggest that night shift work affects the rhythms and levels of circadian Period3 and Nr1d2 expression dependent on the shift schedule or type of the shift; however, there is substantial variation between individuals.

    DOI PubMed

  • Effects of increased daily physical activity on mental health and depression biomarkers in postmenopausal women.

    Takahashi M, Lim PJ, Tsubosaka M, Kim HK, Miyashita M, Suzuki K, Tan EL, Shibata S

    Journal of physical therapy science   31 ( 4 ) 408 - 413  2019.04  [Refereed]  [Domestic journal]

     View Summary

    [Purpose] Little is known about the effectiveness of daily physical activity on depression biomarkers in older adults. This study aimed to investigate the effects of increased daily physical activity for 8 weeks on depression biomarkers in postmenopausal women. [Participants and Methods] Thirty-eight postmenopausal females were randomly assigned into a control or an active group and were asked to wear a uniaxial accelerometer for 8 weeks. Blood samples were obtained at baseline and at the end of the intervention. During the intervention, the active group was asked to increase their physical activity level above their usual lifestyle whereas those in the control group maintained their daily lifestyle. [Results] After the 8-week intervention, the step counts of the participants in the active group increased. The serum concentration of the brain-derived neurotrophic factor and serotonin increased significantly in the active group, but not in the control group, as compared with baseline values. The serum concentration of derivatives of reactive oxygen metabolites and biological antioxidant potential did not change after the intervention in either group. [Conclusion] These findings may suggest that promotion of daily physical activity in postmenopausal women has a positive impact on depression without any change in oxidative stress.

    DOI PubMed

  • Anxiolytic effects of γ-oryzanol in chronically- stressed mice are related to monoamine levels in the brain.

    Akter S, Sasaki H, Uddin KR, Ikeda Y, Miyakawa H, Shibata S

    Life sciences   216   119 - 128  2019.01  [Refereed]  [International journal]

     View Summary

    AIMS: The present study was aimed to investigate the anxiolytic effect of γ-oryzanol (GORZ) during chronic restraint stress treatment (CRST), which is a well-documented model of stress-related disorders, like anxiety, and its potential molecular mechanisms. MATERIALS AND METHODS: In this experiment, 5-week-old male ICR mice were used and the concentration of GORZ was fixed at 0.5% in the mouse standard diet (14% casein, AIN 93 M). Mice were immobilized daily for 3 h from ZT 2.5 to 5.5 (ZT0 was designated as light-on time) for 20 consecutive days, followed by behavioral testing, including the open field test (OFT) and elevated plus maze (EPM) test. The concentration of serum corticosterone (CORT) was measured. In addition, the expression of central monoamine neurotransmitters with their metabolites in the hippocampus, cerebral cortex, and amygdala of the brain were examined. KEY FINDINGS: 0.5% GORZ partially blocked stress-induced reduction of body weight gain while stressed mice had significantly lower body weights during the entire experimental period. Further, 0.5% GORZ treatment could significantly improve the main behavioral parameters even in CRST situations. The significant increase in serum CORT levels indicated CRST-induced stress, which was almost unaffected by 0.5% GORZ treatment. Moreover, 0.5% GORZ also supported the anxiolytic mechanism with enhancement of 5-HIAA and NE levels in the amygdala of brain after CRST. SIGNIFICANCE: Taken together, our studies suggested that 0.5% GORZ is a potential therapeutic drug candidate against anxiety under chronic stress conditions.

    DOI PubMed

  • Effects of Meal Timing on Postprandial Glucose Metabolism and Blood Metabolites in Healthy Adults.

    Masaki Takahashi, Mamiho Ozaki, Moon-Il Kang, Hiroyuki Sasaki, Mayuko Fukazawa, Tamao Iwakami, Pei Jean Lim, Hyeon-Ki Kim, Shinya Aoyama, Shigenobu Shibata

    Nutrients   10 ( 11 )  2018.11  [Refereed]  [International journal]

     View Summary

    We examined the effects of meal timing on postprandial glucose metabolism, including the incretin response and metabolites in healthy adults. Nineteen healthy young men completed two trials involving blood collection in a fasting state and at 30, 60 and 120 min after meal provision in a random order: (1) morning (~0900 h) and (2) evening (~1700 h). The blood metabolome of eight participants was analyzed using capillary electrophoresis-mass spectrometry. Postprandial glucose concentrations at 120 min (p = 0.030) and glucose-dependent insulinotropic polypeptide concentrations (p = 0.005) at 60 min in the evening trials were higher than those in the morning trials. The incremental area under the curve values of five glycolysis, tricarboxylic acid cycle and nucleotide-related metabolites and 18 amino acid-related metabolites were higher in the morning trials than those in the evening trials (p < 0.05). Partial least-squares analysis revealed that the total metabolic change was higher in the morning. Our study demonstrates that a meal in the evening exacerbates the state of postprandial hyperglycemia in healthy adults. In addition, this study provides insight into the difference of incretion and blood metabolites between breakfast and dinner, indicating that the total metabolic responses tends to be higher in the morning.

    DOI PubMed

  • Intracellular-to-total water ratio explains the variability of muscle strength dependence on the size of the lower leg in the elderly.

    Tsukasa Yoshida, Yosuke Yamada, Fumiko Tanaka, Takaki Yamagishi, Shigenobu Shibata, Yasuo Kawakami

    Experimental gerontology   113   120 - 127  2018.11  [Refereed]  [International journal]

     View Summary

    Bioelectrical impedance spectroscopy (BIS) can assess intracellular water (ICW) and total water (TW) in limbs. This study aimed to examine whether BIS can explain a part of the inter-individual variation of the muscle size-strength relationship in older adults. We analyzed the data of 79 participants aged 64-86 years. The maximal voluntary isometric torques of dorsiflexion and plantar flexion on the right side were measured. The anterior and posterior muscle thickness (MT) in the right lower leg was assessed using ultrasonography. The length of the right lower leg (L) was measured, and the ICW-to-TW ratio (ICW/TW) in the right lower leg was obtained using BIS. The MT was multiplied by L to represent an index of muscle volume (MV). Correlation and stepwise regression analyses were performed. The anterior and posterior MT × L significantly and positively correlated with the muscle torque of dorsiflexion and plantar flexion (r = 0.710 and 0.649, respectively, P < 0.001). In the stepwise regression analyses, ICW/TW was selected as a significant predictor of muscle torque independent of MT × L (P < 0.05) for both dorsiflexion and plantar flexion. Electrical parameters of BIS (membrane capacitance, characteristics frequency, and phase angle) in the lower leg also significantly correlated with muscle torques. In addition, the skeletal muscle mass index (appendicular lean mass/height2) was also associated with ICW/TW (P < 0.001). The present results suggest that ICW/TW explains the interindividual variations of the muscle size-strength relationship.

    DOI PubMed

  • Day-Night Oscillation of Atrogin1 and Timing-Dependent Preventive Effect of Weight-Bearing on Muscle Atrophy.

    Shinya Aoyama, Shuichi Kojima, Keisuke Sasaki, Ryosuke Ishikawa, Mizuho Tanaka, Takeru Shimoda, Yuta Hattori, Natsumi Aoki, Kengo Takahashi, Rina Hirooka, Miku Takizawa, Atsushi Haraguchi, Shigenobu Shibata

    EBioMedicine   37   499 - 508  2018.11  [Refereed]  [International journal]

     View Summary

    BACKGROUND: Atrogin1, which is one of the key genes for the promotion of muscle atrophy, exhibits day-night variation. However, its mechanism and the role of its day-night variation are largely unknown in a muscle atrophic context. METHODS: The mice were induced a muscle atrophy by hindlimb-unloading (HU). To examine a role of circadian clock, Wild-type (WT) and Clock mutant mice were used. To test the effects of a neuronal effects, an unilateral ablation of sciatic nerve was performed in HU mice. To test a timing-dependent effects of weight-bearing, mice were released from HU for 4 h in a day at early or late active phase (W-EAP and W-LAP groups, respectively). FINDINGS: We found that the day-night oscillation of Atrogin1 expression was not observed in Clock mutant mice or in the sciatic denervated muscle. In addition, the therapeutic effects of weight-bearing were dependent on its timing with a better effect in the early active phase. INTERPRETATION: These findings suggest that the circadian clock controls the day-night oscillation of Atrogin1 expression and the therapeutic effects of weight-bearing are dependent on its timing. FUND: Council for Science, Technology, and Innovation, SIP, "Technologies for creating next-generation agriculture, forestry, and fisheries".

    DOI PubMed

  • Social jetlag and menstrual symptoms among female university students.

    Komada Y, Ikeda Y, Sato M, Kami A, Masuda C, Shibata S

    Chronobiology international   36 ( 2 ) 1 - 7  2018.11  [Refereed]

    DOI PubMed

  • Circadian-based Treatment Strategy Effective in the BACHD Mouse Model of Huntington's Disease.

    Daniel S Whittaker, Dawn H Loh, Huei-Bin Wang, Yu Tahara, Dika Kuljis, Tamara Cutler, Cristina A Ghiani, Shigenobu Shibata, Gene D Block, Christopher S Colwell

    Journal of biological rhythms   33 ( 5 ) 535 - 554  2018.10  [Refereed]  [International journal]

     View Summary

    Huntington's disease (HD) patients suffer from progressive neurodegeneration that results in cognitive, psychiatric, cardiovascular, and motor dysfunction. Disturbances in sleep-wake cycles are common among HD patients with reports of delayed sleep onset, frequent bedtime awakenings, and excessive fatigue. The BACHD mouse model exhibits many HD core symptoms including circadian dysfunction. Because circadian dysfunction manifests early in the disease in both patients and mouse models, we sought to determine if early interventions that improve circadian rhythmicity could benefit HD symptoms and delay disease progression. We evaluated the effects of time-restricted feeding (TRF) on the BACHD mouse model. At 3 months of age, the animals were divided into 2 groups: ad lib and TRF. The TRF-treated BACHD mice were exposed to a 6-h feeding/18-h fasting regimen that was designed to be aligned with the middle (ZT 15-21) of the period when mice are normally active (ZT 12-24). Following 3 months of treatment (when mice reached the early disease stage), the TRF-treated BACHD mice showed improvements in their locomotor activity and sleep behavioral rhythms. Furthermore, we found improved heart rate variability, suggesting that their autonomic nervous system dysfunction was improved. On a molecular level, TRF altered the phase but not the amplitude of the PER2::LUC rhythms measured in vivo and in vitro. Importantly, treated BACHD mice exhibited improved motor performance compared with untreated BACHD controls, and the motor improvements were correlated with improved circadian output. It is worth emphasizing that HD is a genetically caused disease with no known cure. Lifestyle changes that not only improve the quality of life but also delay disease progression for HD patients are greatly needed. Our study demonstrates the therapeutic potential of circadian-based treatment strategies in a preclinical model of HD.

    DOI PubMed

  • A low-protein diet eliminates the circadian rhythm of serum insulin and hepatic lipid metabolism in mice.

    Yokota SI, Nakamura K, Ando M, Haraguchi A, Omori K, Shibata S

    The Journal of nutritional biochemistry   63   177 - 185  2018.10  [Refereed]

    DOI PubMed

  • Combined effect of shortened photoperiod and low crude protein diet on liver triglyceride accumulation and lipid-related gene expression in quail

    Shin-Ichi Yokota, Midori Ando, Kaai Nakamura, Shigenobu Shibata

    Livestock Science   214   68 - 72  2018.08  [Refereed]

     View Summary

    A short photoperiod is reported to promote body weight gain in quail, and might therefore influence lipid metabolism and induce liver steatosis. Protein deficiency or malnutrition can also cause hepatic steatosis in mammals, but the effect in birds has not previously been investigated. In order to determine the influence of a shortened photoperiod and low crude protein diet on avian hepatic lipid metabolism, 10-day-old male Japanese quail (Coturnix coturnix japonica) were assigned to a 2 × 2 factorial arrangement of treatments in four groups, each of eight birds, and fed for three days, after which liver triglyceride (TG) accumulation and lipid-related gene expression were measured. Experimental factors included two dietary protein levels of 24% or 10% crude protein (CP), and two photoperiod conditions of a 12:12 h light/dark cycle (12L:12D) or a 1:23 h light/dark cycle (1L:23D). The liver TG level in quail fed the 10% CP diet under 1L:23D was higher than in birds maintained on either the 10% CP diet under 12L:12D (15.86 vs. 10.56 mg/g liver, P &lt
    0.01) or 24% CP diet under 1L:23D (7.95 mg/g liver, P &lt
    0.01). The sterol regulatory element-binding protein 1 mRNA, which is involved in TG synthesis, was more highly expressed in quail on the 10% CP diet under 1L:23D than in those maintained on the 24% CP diet under 12L:12D (2.2 vs. 7.9, P &lt
    0.01), whereas the mRNA level of very low-density apolipoprotein II (apoVLDLII), which encodes a VLDL structural protein, was lower (157.3 vs. 12.5, P &lt
    0.01). To our knowledge, this is the first report of a selective protein restriction and shorter photoperiod, alone and in combination, inducing a fatty liver state in an avian species. Our study provides insight into factors that govern lipid metabolism in birds, and the efficiency of domestic fowl production.

    DOI

  • Chronotype and social jetlag influence human circadian clock gene expression.

    Masaki Takahashi, Yu Tahara, Miku Tsubosaka, Mayuko Fukazawa, Mamiho Ozaki, Tamao Iwakami, Takashi Nakaoka, Shigenobu Shibata

    Scientific reports   8 ( 1 ) 10152 - 10152  2018.07  [Refereed]  [International journal]

     View Summary

    We examined the relationships between chronotype or social jetlag and clock gene expression. Twenty-four young men [Chronotype: morningness, n = 8; intermediate, n = 8, eveningness, n = 8], aged 27 ± 2 years old (mean ± SE), completed two trials in a randomized order: (1) a Friday trial and (2) a Monday trial. In both trials, hair follicle cells were collected to evaluate the expression of clock genes over a 24-hour period at 4-hour intervals. There was a significant main effect of time on the expression of NR1D1, NR1D2, and PER3 (P < 0.001) in the morningness group, but not in the eveningness group. Changes in the peak time of expression of NR1D1 (r = 0.434, P = 0.034), NR1D2 (r = 0.481, P = 0.017), and PER3 (r = 0.457, P = 0.025) from the Friday to Monday trials were positively correlated with social jetlag (SJL) time. Our findings indicate that there was no change in the patterns of clock gene expression between workdays and the day after the holiday in the morningness group, and that SJL time influences the peak time of clock gene expression, moving it from the early to late workday, after a holiday.

    DOI PubMed

  • A randomized, double-blind and placebo-controlled crossover trial on the effect of l-ornithine ingestion on the human circadian clock.

    Fukuda T, Haraguchi A, Takahashi M, Nakaoka T, Fukazawa M, Okubo J, Ozaki M, Kanatome A, Ohya R, Miura Y, Obara K, Shibata S

    Chronobiology international   35 ( 10 ) 1 - 11  2018.07  [Refereed]

    DOI PubMed

  • Circadian clock component PERIOD2 regulates diurnal expression of Na+/H+ exchanger regulatory factor-1 and its scaffolding function.

    Yuya Tsurudome, Satoru Koyanagi, Takumi Kanemitsu, Chiharu Katamune, Masayuki Oda, Yuki Kanado, Mizuki Kato, Akari Morita, Yu Tahara, Naoya Matsunaga, Shigenobu Shibata, Shigehiro Ohdo

    Scientific reports   8 ( 1 ) 9072 - 9072  2018.06  [Refereed]  [International journal]

     View Summary

    A number of diverse cell-surface proteins are anchored to the cytoskeleton via scaffold proteins. Na+/H+ exchanger regulatory factor-1 (NHERF1), encoded by the Slc9a3r1 gene, functions as a scaffold protein, which is implicated in the regulation of membrane expression of various cell-surface proteins. Here, we demonstrate that the circadian clock component PERIOD2 (PER2) modulates transcription of the mouse Slc9a3r1 gene, generating diurnal accumulation of NHERF1 in the mouse liver. Basal expression of Slc9a3r1 was dependent on transcriptional activation by p65/p50. PER2 bound to p65 protein and prevented p65/p50-mediated transactivation of Slc9a3r1. The time-dependent interaction between PER2 and p65 underlay diurnal oscillation in the hepatic expression of Slc9a3r1/NHERF1. The results of immunoprecipitation experiments and liquid chromatography-mass spectrometry analysis of mouse liver revealed that NHERF1 time-dependently interacted with fatty acid transport protein-5 (FATP5). Temporary accumulation of NHERF1 protein stabilized plasmalemmal localization of FATP5, thereby enhancing hepatic uptake of fatty acids at certain times of the day. Our results suggest an unacknowledged role for PER2 in regulating the diurnal expression of NHERF1 in mouse liver. This machinery also contributed to diurnal changes in the ability of hepatic cells to uptake fatty acids.

    DOI PubMed

  • γ-oryzanol ameliorates the acute stress induced by behavioral anxiety testing in mice.

    Akter S, Sasaki H, Ikeda Y, Miyakawa H, Shibata S

    Journal of pharmacological sciences   138 ( 2 ) 155 - 159  2018.06  [Refereed]  [Domestic journal]

     View Summary

    We evaluated the anxiolytic effect of γ-oryzanol (GORZ) and elucidated the molecular mechanisms involved in its inhibition of behavioral test-induced anxiety. Behavioral tests were conducted on day 13, and mice were subjected to 30 min of acute restraint stress treatment (ARST) before sacrifice on day 16. In other group, behavioral tests were conducted on day 13 and 14 after ARST. 0.5% GORZ significantly weakened the effect of behavioral stress, but not the effect of strong ARST. GORZ downregulated ARST-induced cFos levels in the cerebral cortex. In conclusion, GORZ has potential ant-anxiety effect in the treatment of weak behavioral test-induced stress.

    DOI PubMed

  • Entrainment of the mouse circadian clock: Effects of stress, exercise, and nutrition.

    Yu Tahara, Shigenobu Shibata

    Free radical biology & medicine   119   129 - 138  2018.05  [Refereed]  [International journal]

     View Summary

    The circadian clock system in mammals plays a fundamental role in maintaining homeostasis. Entrainment is an important characteristic of the internal clock, by which appropriate timing is maintained according to external daily stimuli, such as light, stress, exercise, and/or food. Disorganized entrainment or a misaligned clock time, such as jet lag, increases health disturbances. The central clock in the suprachiasmatic nuclei, located in the hypothalamus, receives information about arousal stimuli, such as physical stress or exercise, and changes the clock time by modifying neural activity or the expression of circadian clock genes. Although feeding stimuli cannot entrain the central clock in a normal light-dark cycle, the central clock can partially detect the metabolic status. Local clocks in the peripheral tissues, including liver and kidney, have a strong direct response to the external stimuli of stress, exercise, and/or food that is independent of the central clock. The mechanism underlying entrainment by stress/exercise is mediated by glucocorticoids, sympathetic nerves, oxidative stress, hypoxia, pH, cytokines, and temperature. Food/nutrition-induced entrainment is mediated by fasting-induced hormonal or metabolic changes and re-feeding-induced insulin or oxyntomodulin secretion. Chrono-nutrition is a clinical application based on chronobiology research. Future studies are required to elucidate the effects of eating and nutrient composition on the human circadian clock. Here, we focus on the central and peripheral clocks mostly in rodents' studies and review the findings of recent investigations of the effects of stress, exercise, and food on the entrainment system.

    DOI PubMed

  • 医療のさらなる安心・安全を志向する若手研究者のイノベーティブチャレンジ 時計遺伝子Per2の発現周期やマウス行動周期に対する遠志の影響について

    原口 敦嗣, 中村 文彬, 阿部 真太郎, 中尾 洋一, 柴田 重信

    日本薬学会年会要旨集   138年会 ( 1 ) 336 - 336  2018.03

  • Glucagon and/or IGF-1 Production Regulates Resetting of the Liver Circadian Clock in Response to a Protein or Amino Acid-only Diet.

    Ikeda Y, Kamagata M, Hirao M, Yasuda S, Iwami S, Sasaki H, Tsubosaka M, Hattori Y, Todoh A, Tamura K, Shiga K, Ohtsu T, Shibata S

    EBioMedicine   28   210 - 224  2018.02  [Refereed]  [International journal]

     View Summary

    The circadian system controls the behavior and multiple physiological functions. In mammals, the suprachiasmatic nucleus (SCN) acts as the master pacemaker and regulates the circadian clocks of peripheral tissues. The SCN receives information regarding the light-dark cycle and is thus synchronized to the external 24-hour environment. In contrast, peripheral clocks, such as the liver clock, receive information from the SCN and other factors; in particular, food intake which leads to insulin secretion induces strong entrainment of the liver clock. On the other hand, the liver clock of insulin-depleted mice treated with streptozotocin (STZ) has been shown to be entrained by scheduled feeding, suggesting that insulin is not necessary for entrainment of the liver clock by feeding. In this study, we aimed to elucidate additional mechanism on entraining liver clock by feeding a protein-only diet and/or amino-acid administration which does not increase insulin levels. We demonstrated that protein-only diet and cysteine administration elicit entrainment of the liver clock via glucagon secretion and/or insulin-like growth factors (IGF-1) production. Our findings suggest that glucagon and/or IGF-1 production are additional key factors in food-induced entrainment.

    DOI PubMed

  • Gut Microbiota-Derived Short Chain Fatty Acids Induce Circadian Clock Entrainment in Mouse Peripheral Tissue.

    Yu Tahara, Mayu Yamazaki, Haruna Sukigara, Hiroaki Motohashi, Hiroyuki Sasaki, Hiroki Miyakawa, Atsushi Haraguchi, Yuko Ikeda, Shinji Fukuda, Shigenobu Shibata

    Scientific reports   8 ( 1 ) 1395 - 1395  2018.01  [Refereed]  [International journal]

     View Summary

    Microbiota-derived short-chain fatty acids (SCFAs) and organic acids produced by the fermentation of non-digestible fibre can communicate from the microbiome to host tissues and modulate homeostasis in mammals. The microbiome has circadian rhythmicity and helps the host circadian clock function. We investigated the effect of SCFA or fibre-containing diets on circadian clock phase adjustment in mouse peripheral tissues (liver, kidney, and submandibular gland). Initially, caecal SCFA concentrations, particularly acetate and butyrate, induced significant day-night differences at high concentrations during the active period, which were correlated with lower caecal pH. By monitoring luciferase activity correlated with the clock gene Period2 in vivo, we found that oral administration of mixed SCFA (acetate, butyrate, and propionate) and an organic acid (lactate), or single administration of each SCFA or lactate for three days, caused phase changes in the peripheral clocks with stimulation timing dependency. However, this effect was not detected in cultured fibroblasts or cultured liver slices with SCFA applied to the culture medium, suggesting SCFA-induced indirect modulation of circadian clocks in vivo. Finally, cellobiose-containing diets facilitated SCFA production and refeeding-induced peripheral clock entrainment. SCFA oral gavage and prebiotic supplementation can facilitate peripheral clock adjustment, suggesting prebiotics as novel therapeutic candidates for misalignment.

    DOI PubMed

  • Night eating model shows time-specific depression-like behavior in the forced swimming test.

    Atsushi Haraguchi, Miyabi Fukuzawa, Shiho Iwami, Yutaro Nishimura, Hiroaki Motohashi, Yu Tahara, Shigenobu Shibata

    Scientific reports   8 ( 1 ) 1081 - 1081  2018.01  [Refereed]  [International journal]

     View Summary

    The circadian clock system is associated with feeding and mood. Patients with night eating syndrome (NES) delay their eating rhythm and their mood declines during the evening and night, manifesting as time-specific depression. Therefore, we hypothesized that the NES feeding pattern might cause time-specific depression. We established new NES model by restricted feeding with high-fat diet during the inactive period under normal-fat diet ad libitum. The FST (forced swimming test) immobility time in the NES model group was prolonged only after lights-on, corresponding to evening and early night for humans. We examined the effect of the NES feeding pattern on peripheral clocks using PER2::LUCIFERASE knock-in mice and an in vivo monitoring system. Caloric intake during the inactive period would shift the peripheral clock, and might be an important factor in causing the time-specific depression-like behavior. In the NES model group, synthesis of serotonin and norepinephrine were increased, but utilization and metabolism of these monoamines were decreased under stress. Desipramine shortened some mice's FST immobility time in the NES model group. The present study suggests that the NES feeding pattern causes phase shift of peripheral clocks and malfunction of the monoamine system, which may contribute to the development of time-specific depression.

    DOI PubMed

  • Chrono-nutrition and n-3 polyunsaturated fatty acid

    Shigenobu Shibata, Akiko Furutani

    Folia Pharmacologica Japonica   151 ( 1 ) 34 - 40  2018  [Refereed]

     View Summary

    Circadian clock system has been widely maintained in many spices from prokaryote to mammals. “Circadian” means “approximately day” in Latin, thus circadian rhythm means about 24 hour rhythms. The earth revolves once every 24 hours, and our circadian system has been developed for adjusting to this 24 hour cycles, to get sun light information for getting their foods or for alive in birds or mammals. We have two different circadian systems so-called main oscillator located in the suprachiasmatic nucleus (SCN) of the hypothalamus, and local oscillator located in the various peripheral organ tissues such as liver, kidney and skeletal muscle. The SCN is directly entrained by light-dark information through retinal-hypothalamic tract, and then organizes local clock in peripheral tissues via many pathways including neural and hormonal functions. On the other hand, peripheral local clocks are entrained by feeding, exercise and stress stimuli through several cell signaling. Foods (protein, carbohydrate, and lipid) are important regulator of circadian clocks in peripheral tissues. Thus, controlling the timing of food consumption and food composition, a concept known as chrononutrition, is one area of active research to aid recovery from many physiological dysfunctions. In this review, we focus on molecular mechanisms of entrainment and the relationships between circadian clock systems and n-3 polyunsaturated fatty acid. We concentrate on experimental data obtained from cells or animals and humans and discuss how these findings translate into clinical research, and we highlight the latest developments in chrononutritional studies.

    DOI PubMed

  • Association of body mass index-related single nucleotide polymorphisms with psychiatric disease and memory performance in a Japanese population

    Midori Ninomiya-Baba, Junko Matsuo, Daimei Sasayama, Hiroaki Hori, Toshiya Teraishi, Miho Ota, Kotaro Hattori, Takamasa Noda, Ikki Ishida, Shigenobu Shibata, Hiroshi Kunugi

    Acta Neuropsychiatrica   29 ( 5 ) 299 - 308  2017.10  [Refereed]

     View Summary

    Objective Obesity is a risk factor for psychiatric diseases. Recently, a number of single nucleotide polymorphisms (SNPs) have been shown to be related to body mass index (BMI). In this study, we investigated the association of BMI-related SNPs with psychiatric diseases and one of their endophenotypes, memory performance, in a Japanese population. Methods The subjects were 1624 patients with one of three psychiatric diseases (799 patients with major depressive disorder, 594 with schizophrenia, and 231 with bipolar disorder) and 1189 healthy controls. Memory performance was assessed using the Wechsler Memory Scale-Revised (WMS-R). Genomic DNA was prepared from venous blood and used to genotype 23 BMI-related SNPs using the TaqMan 5′-exonuclease allelic discrimination assay. We then analysed the relationships between the SNPs and psychiatric disease and various subscales of the WMS-R. Results Three SNPs (rs11142387, rs12597579, and rs6548238) showed significant differences in the genotype or allele frequency between patients with any psychiatric diseases and controls. Furthermore, six SNPs (rs11142387, rs12597579, rs2815752, rs2074356, rs4776970, and rs2287019) showed significant differences in at least one subscale of the WMS-R depending on the genotypes of the healthy controls. Interestingly, rs11142387 near the Kruppel-like factor 9 (KLF9) was significantly associated with psychiatric disease and poor memory function. Conclusions We identified three and six BMI-related SNPs associated with psychiatric disease and memory performance, respectively. In particular, carrying the A allele of rs11142387 near KLF9 was found to be associated with psychiatric disease and poor memory performance, which warrants further investigations.

    DOI PubMed

  • Association of body mass nindex-related single nucleotide polymorphisms with psychiatric disease and memory performance in a Japanese population

    Midori Ninomiya-Baba, Junko Matsuo, Daimei Sasayama, Hiroaki Hori, Toshiya Teraishi, Miho Ota, Kotaro Hattori, Takamasa Noda, Ikki Ishida, Shigenobu Shibata, Hiroshi Kunugi

    ACTA NEUROPSYCHIATRICA   29 ( 5 ) 299 - 308  2017.10  [Refereed]

     View Summary

    Objective: Obesity is a risk factor for psychiatric diseases. Recently, a number of single nucleotide polymorphisms (SNPs) have been shown to be related to body mass index (BMI). In this study, we investigated the association of BMI-related SNPs with psychiatric diseases and one of their endophenotypes, memory performance, in a Japanese population.
    Methods: The subjects were 1624 patients with one of three psychiatric diseases (799 patients with major depressive disorder, 594 with schizophrenia, and 231 with bipolar disorder) and 1189 healthy controls. Memory performance was assessed using the Wechsler Memory Scale Revised (WMS-R). Genomic DNA was prepared from venous blood and used to genotype 23 BMI-related SNPs using the TaqMan 5'-exonuclease allelic discrimination assay. We then analysed the relationships between the SNPs and psychiatric disease and various subscales of the WMS-R.
    Results: Three SNPs (rs11142387, rs12597579, and rs6548238) showed significant differences in the genotype or allele frequency between patients with any psychiatric diseases and controls. Furthermore, six SNPs (rs11142387, rs12597579, rs2815752, rs2074356, rs4776970, and rs2287019) showed significant differences in at least one subscale of the WMS-R depending on the genotypes of the healthy controls. Interestingly, rs11142387 near the Kruppel-like factor 9 (KLF9) was significantly associated with psychiatric disease and poor memory function.
    Conclusions: We identified three and six BMI-related SNPs associated with psychiatric disease and memory performance, respectively. In particular, carrying the A allele of rs11142387 near KLF9 was found to be associated with psychiatric disease and poor memory performance, which warrants further investigations.

    DOI

  • Age-dependent motor dysfunction due to neuron-specific disruption of stress-activated protein kinase MKK7

    Tokiwa Yamasaki, Norie Deki-Arima, Asahito Kaneko, Norio Miyamura, Mamiko Iwatsuki, Masato Matsuoka, Noriko Fujimori-Tonou, Yoshimi Okamoto-Uchida, Jun Hirayama, Jamey D. Marth, Yuji Yamanashi, Hiroshi Kawasaki, Koji Yamanaka, Josef M. Penninger, Shigenobu Shibata, Hiroshi Nishina

    SCIENTIFIC REPORTS   7 ( 1 ) 7348  2017.08  [Refereed]

     View Summary

    c-Jun N-terminal kinase (JNK) is a member of the mitogen-activated protein kinase family and controls various physiological processes including apoptosis. A specific upstream activator of JNKs is the mitogen-activated protein kinase kinase 7 (MKK7). It has been reported that MKK7-JNK signaling plays an important regulatory role in neural development, however, post-developmental functions in the nervous system have not been elucidated. In this study, we generated neuron-specific Mkk7 knockout mice (MKK7 cKO), which impaired constitutive activation of JNK in the nervous system. MKK7 cKO mice displayed impaired circadian behavioral rhythms and decreased locomotor activity. MKK7 cKO mice at 8 months showed motor dysfunctions such as weakness of hind-limb and gait abnormality in an age-dependent manner. Axonal degeneration in the spinal cord and muscle atrophy were also observed, along with accumulation of the axonal transport proteins JNK-interacting protein 1 and amyloid beta precursor protein in the brains and spinal cords of MKK7 cKO mice. Thus, the MKK7-JNK signaling pathway plays important roles in regulating circadian rhythms and neuronal maintenance in the adult nervous system.

    DOI PubMed

  • Circadian clock-dependent increase in salivary IgA secretion modulated by sympathetic receptor activation in mice

    Misaki Wada, Kanami Orihara, Mayo Kamagata, Koki Hama, Hiroyuki Sasaki, Atsushi Haraguchi, Hiroki Miyakawa, Atsuhito Nakao, Shigenobu Shibata

    SCIENTIFIC REPORTS   7 ( 1 ) 8802  2017.08  [Refereed]

     View Summary

    The salivary gland is rhythmically controlled by sympathetic nerve activation from the suprachiasmatic nucleus (SCN), which functions as the main oscillator of circadian rhythms. In humans, salivary IgA concentrations reflect circadian rhythmicity, which peak during sleep. However, the mechanisms controlling this rhythmicity are not well understood. Therefore, we examined whether the timing of parasympathetic (pilocarpine) or sympathetic (norepinephrine; NE) activation affects IgA secretion in the saliva. The concentrations of saliva IgA modulated by pilocarpine activation or by a combination of pilocarpine and NE activation were the highest in the middle of the light period, independent of saliva flow rate. The circadian rhythm of IgA secretion was weakened by an SCN lesion and Clock gene mutation, suggesting the importance of the SCN and Clock gene on this rhythm. Adrenoceptor antagonists blocked both NE- and pilocarpine-induced basal secretion of IgA. Dimeric IgA binds to the polymeric immunoglobulin receptor (pIgR) on the basolateral surface of epithelial cells and forms the IgA-pIgR complex. The circadian rhythm of Pigr abundance peaked during the light period, suggesting pIgR expression upon rhythmic secretion of IgA. We speculate that activation of sympathetic nerves during sleep may protect from bacterial access to the epithelial surface through enhanced secretion of IgA.

    DOI PubMed

  • Clock-dependent temporal regulation of IL-33/ST2-mediated mast cell response

    Takahiro Kawauchi, Kayoko Ishimaru, Yuki Nakamura, Nobuhiro Nakano, Mutsuko Hara, Hideoki Ogawa, Ko Okumura, Shigenobu Shibata, Atsuhito Nakao

    ALLERGOLOGY INTERNATIONAL   66 ( 3 ) 472 - 478  2017.07  [Refereed]

     View Summary

    Background: Interleukin-33 (IL-33) is an alarmin cytokine that binds to the interleukin 1 receptor-like 1 protein ST2. Clock is a key circadian gene that is essential for endogenous clockworks in mammals. This study investigated whether Clock temporally regulated IL-33-mediated responses in mast cells.
    Methods: The kinetics of IL-33-mediated IL-6, IL-13, and TNF-alpha productions were compared between bone marrow-derived mast cells (BMMCs) from wild-type and Clock-mutated mice (Clock(Delta 19/Delta 19) mice). The kinetics of the neutrophil influx into the peritoneal cavity or expression of IL-13 and Gob-5 in the lung in response to IL-33 were compared between wild-type and Clock(Delta 19/Delta 19) mice. We also examined the kinetics of ST2 expression in mast cells and its association with Clock expression.
    Results: There was a time-of-day-dependent variation in IL-33-mediated IL-6, IL-13, and TNF-alpha production in wild-type BMMCs, which was absent in Clock-mutated BMMCs. IL-33-induced neutrophil infiltration into the peritoneal cavity also showed a time-of-day-dependent variation in wild-type mice, which was absent in Clock(Delta 19/Delta 19) mice. Furthermore, IL-33-induced IL-13 and Gob-5 expression in the lung exhibited a time-of-day-dependent variation in wild-type mice. These temporal variations in IL-33 mediated mast cell responses were associated with temporal variations of ST2 expression in mast cells. In addition, CLOCK bound to the promoter region of ST2 and Clock deletion resulted in down-regulation of ST2 expression in mast cells.
    Conclusions: CLOCK temporally gates mast cell responses to IL-33 via regulation of ST2 expression. Our findings provide novel insights into IL-33/mast cell-associated physiology and pathologies. Copyright (C) 2017, Japanese Society of Allergology. Production and hosting by Elsevier B.V.

    DOI PubMed

  • Polyporus and Bupleuri radix effectively alter peripheral circadian clock phase acutely in male mice.

    Hiroaki Motohashi, Haruna Sukigara, Yu Tahara, Keisuke Saito, Mayu Yamazaki, Takuya Shiraishi, Yosuke Kikuchi, Atsushi Haraguchi, Shigenobu Shibata

    Nutrition research (New York, N.Y.)   43   16 - 24  2017.07  [Refereed]  [International journal]

     View Summary

    In mammals, daily physiological events are precisely regulated by an internal circadian clock system. An important function of this system is to readjust the phase of the clock daily. In Japan, traditional herb medicines, so-called crude drugs (Shoyaku), are widely used for many diseases, and some are reported to affect circadian clock impairment, suggesting that some of them might have an ability to modify clock gene expression rhythms. Therefore, from selected 40 crude drugs, finding candidates that control the circadian clock phases was the first purpose of this study. As there are several crude drugs used for liver- and/or kidney-related diseases, the second aim of the present study was to find some crude drugs affecting liver/kidney circadian clock in vivo. To assess phase changes in the daily circadian rhythm, bioluminescence from the core clock gene product Period 2 was continuously monitored in mouse embryonic fibroblasts in vitro and in some peripheral tissues (kidney, liver, and submandibular gland) of PERIOD2::LUCIFERASE knock-in mice in vivo. In our screening, Polyporus and Bupleuri radix were found to be good candidates to effectively manipulate the peripheral circadian clock phase acutely, with stimulation time-of-day dependency in vitro as well as in vivo. Interestingly, Polyporus and Bupleuri radix are traditional herb medicines use for treating edema and promoting diuresis, and for chronic hepatitis, respectively. These crude drugs may be therefore good modulators of the circadian peripheral clocks including liver and kidney, and circadian clock genes become new molecular targets for these crude drugs.

    DOI PubMed

  • Abnormal tuning of the hepatic circadian metabolic rhythms in lung cancer.

    Yu Tahara, Shigenobu Shibata

    Hepatology (Baltimore, Md.)   65 ( 3 ) 1061 - 1064  2017.03  [Refereed]  [International journal]

    DOI PubMed

  • Positive association between physical activity and PER3 expression in older adults.

    Masaki Takahashi, Atsushi Haraguchi, Yu Tahara, Natsumi Aoki, Mayuko Fukazawa, Kumpei Tanisawa, Tomoko Ito, Takashi Nakaoka, Mitsuru Higuchi, Shigenobu Shibata

    Scientific reports   7   39771 - 39771  2017.01  [Refereed]  [International journal]

     View Summary

    The circadian clock regulates many physiological functions including physical activity and feeding patterns. In addition, scheduled exercise and feeding themselves can affect the circadian clock. The purpose of the present study was to investigate the relationship between physical/feeding activity and expression of clock genes in hair follicle cells in older adults. Twenty adult men (age, 68 ± 7 years, mean ± SE) were examined in this cross-sectional study. Prior to hair follicle cell collection, the participants were asked to wear a uniaxial accelerometer for one week. The timings of breakfast, lunch, and dinner were also recorded. Hair follicle cells were then collected over a 24 h period at 4 h intervals. The amplitude of PER3 expression was positively correlated with moderate and vigorous physical activity (r = 0.582, p = 0.007) and peak oxygen uptake (r = 0.481, p = 0.032), but these correlations were not observed for NR1D1 or NR1D2. No association was noted between meal times and the amplitude or the acrophase for any of these three clock genes. These findings suggest that rhythmic expression of the circadian clock gene PER3 is associated with the amount of daily physical activity and physical fitness in older adults.

    DOI PubMed

  • The mammalian circadian clock and its entrainment by stress and exercise.

    Yu Tahara, Shinya Aoyama, Shigenobu Shibata

    The journal of physiological sciences : JPS   67 ( 1 ) 1 - 10  2017.01  [Refereed]  [Domestic journal]

     View Summary

    The mammalian circadian clock regulates day-night fluctuations in various physiological processes. The circadian clock consists of the central clock in the suprachiasmatic nucleus of the hypothalamus and peripheral clocks in peripheral tissues. External environmental cues, including light/dark cycles, food intake, stress, and exercise, provide important information for adjusting clock phases. This review focuses on stress and exercise as potent entrainment signals for both central and peripheral clocks, especially in regard to the timing of stimuli, types of stressors/exercises, and differences in the responses of rodents and humans. We suggest that the common signaling pathways of clock entrainment by stress and exercise involve sympathetic nervous activation and glucocorticoid release. Furthermore, we demonstrate that physiological responses to stress and exercise depend on time of day. Therefore, using exercise to maintain the circadian clock at an appropriate phase and amplitude might be effective for preventing obesity, diabetes, and cardiovascular disease.

    DOI PubMed

  • Regulation of plasma histamine levels by the mast cell clock and its modulation by stress

    Yuki Nakamura, Kayoko Ishimaru, Shigenobu Shibata, Atsuhito Nakao

    SCIENTIFIC REPORTS   7   39934  2017.01  [Refereed]

     View Summary

    At steady state, plasma histamine levels exhibit circadian variations with nocturnal peaks, which is implicated in the nighttime exacerbation of allergic symptoms. However, the regulatory mechanisms are largely unexplored. This study determined how steady-state plasma histamine levels are regulated and affected by environmental factors. We found that plasma histamine levels decreased in mast cell-deficient mice and their circadian variations were lost in mast cell-deficient mice reconstituted with bone marrow-derived mast cells (BMMCs) harboring a mutation in the circadian gene Clock. Clock temporally regulates expression of organic cation transporter 3 (OCT3), which is involved in histamine transport, in mast cells; OCT inhibition abolished circadian variations in plasma histamine levels. Mice housed under aberrant light/dark conditions or suffering from restraint stress exhibited desynchronization of the mast cell clockwork, concomitant with the loss of circadian variations in OCT3 expression and plasma histamine levels. The degree of compound 48/80-induced plasma extravasation in mice was correlated with plasma histamine levels. Collectively, the mast cell clock mediates circadian regulation of plasma histamine levels at steady state, in part by controlling OCT3 expression, which can be modulated by stress. Additionally, we propose that plasma histamine levels potentiate mast cell-mediated allergic reactions.

    DOI PubMed

  • Potent Effects of Flavonoid Nobiletin on Amplitude, Period, and Phase of the Circadian Clock Rhythm in PER2::LUCIFERASE Mouse Embryonic Fibroblasts.

    Ayako Shinozaki, Kenichiro Misawa, Yuko Ikeda, Atsushi Haraguchi, Mayo Kamagata, Yu Tahara, Shigenobu Shibata

    PloS one   12 ( 2 ) e0170904  2017  [Refereed]  [International journal]

     View Summary

    Flavonoids are natural polyphenols that are widely found in plants. The effects of flavonoids on obesity and numerous diseases such as cancer, diabetes, and Alzheimer's have been well studied. However, little is known about the relationships between flavonoids and the circadian clock. In this study, we show that continuous or transient application of flavonoids to the culture medium of embryonic fibroblasts from PER2::LUCIFERASE (PER2::LUC) mice induced various modifications in the circadian clock amplitude, period, and phase. Transient application of some of the tested flavonoids to cultured cells induced a phase delay of the PER2::LUC rhythm at the down slope phase. In addition, continuous application of the polymethoxy flavonoids nobiletin and tangeretin increased the amplitude and lengthened the period of the PER2::LUC rhythm. The nobiletin-induced phase delay was blocked by co-treatment with U0126, an ERK inhibitor. In summary, among the tested flavonoids, polymethoxy flavones increased the amplitude, lengthened the period, and delayed the phase of the PER2::LUC circadian rhythm. Therefore, foods that contain polymethoxy flavones may have beneficial effects on circadian rhythm disorders and jet lag.

    DOI PubMed

  • The Role of Circadian Rhythms in Muscular and Osseous Physiology and Their Regulation by Nutrition and Exercise.

    Shinya Aoyama, Shigenobu Shibata

    Frontiers in neuroscience   11   63 - 63  2017  [Refereed]  [International journal]

     View Summary

    The mammalian circadian clock regulates the day and night cycles of various physiological functions. The circadian clock system consists of a central clock in the suprachiasmatic nucleus (SCN) of the hypothalamus and peripheral clocks in peripheral tissues. According to the results of circadian transcriptomic studies in several tissues, the majority of rhythmic genes are expressed in a tissue-specific manner and are influenced by tissue-specific circadian rhythms. Here we review the diurnal variations of musculoskeletal functions and discuss the impact of the circadian clock on homeostasis in skeletal muscle and bone. Peripheral clocks are controlled by not only photic stimulation from the central clock in the SCN but also by external cues, such as feeding and exercise. In this review, we discuss the effects of feeding and exercise on the circadian clock and diurnal variation of musculoskeletal functions. We also discuss the therapeutic potential of chrono-nutrition and chrono-exercise on circadian disturbances and the failure of homeostasis in skeletal muscle and bone.

    DOI PubMed

  • Potent synchronization of peripheral circadian clocks by glucocorticoid injections in PER2::LUC-Clock/Clock mice.

    Mayo Kamagata, Yuko Ikeda, Hiroyuki Sasaki, Yuta Hattori, Shinnosuke Yasuda, Shiho Iwami, Miku Tsubosaka, Ryosuke Ishikawa, Ai Todoh, Konomi Tamura, Yu Tahara, Shigenobu Shibata

    Chronobiology international   34 ( 8 ) 1067 - 1082  2017  [Refereed]  [International journal]

     View Summary

    In mammals, the central clock (the suprachiasmatic nuclei, SCN) is entrained mainly by the light-dark cycle, whereas peripheral clocks in the peripheral tissues are entrained/synchronized by multiple factors, including feeding patterns and endocrine hormones such as glucocorticoids. Clock-mutant mice (Clock/Clock), which have a mutation in a core clock gene, show potent phase resetting in response to light pulses compared with wild-type (WT) mice, owing to the damped and flexible oscillator in the SCN. However, the phase resetting of the peripheral clocks in Clock/Clock mice has not been elucidated. Here, we characterized the peripheral clock gene synchronization in Clock/Clock mice by daily injections of a synthetic glucocorticoid (dexamethasone, DEX) by monitoring in vivo PER2::LUCIFERASE bioluminescence. Compared with WT mice, the Clock/Clock mice showed significantly decreased bioluminescence and peripheral clock rhythms with decreased amplitudes and delayed phases. In addition, the DEX injections increased the amplitudes and advanced the phases. In order to examine the robustness of the internal oscillator, T-cycle experiments involving DEX stimulations with 24- or 30-h intervals were performed. The Clock/Clock mice synchronized to the 30-h T-cycle stimulation, which suggested that the peripheral clocks in the Clock/Clock mice had increased synchronizing ability upon DEX stimulation, to that of circadian and hour-glass type oscillations, because of weak internal clock oscillators.

    DOI PubMed

  • Age-related circadian disorganization caused by sympathetic dysfunction in peripheral clock regulation.

    Yu Tahara, Yuta Takatsu, Takuya Shiraishi, Yosuke Kikuchi, Mayu Yamazaki, Hiroaki Motohashi, Aya Muto, Hiroyuki Sasaki, Atsushi Haraguchi, Daisuke Kuriki, Takahiro J Nakamura, Shigenobu Shibata

    NPJ aging and mechanisms of disease   3   16030 - 16030  2017  [Refereed]  [International journal]

     View Summary

    The ability of the circadian clock to adapt to environmental changes is critical for maintaining homeostasis, preventing disease, and limiting the detrimental effects of aging. To date, little is known about age-related changes in the entrainment of peripheral clocks to external cues. We therefore evaluated the ability of the peripheral clocks of the kidney, liver, and submandibular gland to be entrained by external stimuli including light, food, stress, and exercise in young versus aged mice using in vivo bioluminescence monitoring. Despite a decline in locomotor activity, peripheral clocks in aged mice exhibited normal oscillation amplitudes under light-dark, constant darkness, and simulated jet lag conditions, with some abnormal phase alterations. However, age-related impairments were observed in peripheral clock entrainment to stress and exercise stimuli. Conversely, age-related enhancements were observed in peripheral clock entrainment to food stimuli and in the display of food anticipatory behaviors. Finally, we evaluated the hypothesis that deficits in sympathetic input from the central clock located in the suprachiasmatic nucleus of the hypothalamus were in part responsible for age-related differences in the entrainment. Aged animals showed an attenuated entrainment response to noradrenergic stimulation as well as decreased adrenergic receptor mRNA expression in target peripheral organs. Taken together, the present findings indicate that age-related circadian disorganization in entrainment to light, stress, and exercise is due to sympathetic dysfunctions in peripheral organs, while meal timing produces effective entrainment of aged peripheral circadian clocks.

    DOI PubMed

  • L-Ornithine affects peripheral clock gene expression in mice

    Takafumi Fukuda, Atsushi Haraguchi, Mari Kuwahara, Kaai Nakamura, Yutaro Hamaguchi, Yuko Ikeda, Yuko Ishida, Guanying Wang, Chise Shirakawa, Yoko Tanihata, Kazuaki Ohara, Shigenobu Shibata

    SCIENTIFIC REPORTS   6   34665  2016.10  [Refereed]

     View Summary

    The peripheral circadian clock is entrained by factors in the external environment such as scheduled feeding, exercise, and mental and physical stresses. In addition, recent studies in mice demonstrated that some food components have the potential to control the peripheral circadian clock during scheduled feeding, although information about these components remains limited. L-Ornithine is a type of non-protein amino acid that is present in foods and has been reported to have various physiological functions. In human trials, for example, L-ornithine intake improved a subjective index of sleep quality. Here we demonstrate, using an in vivo monitoring system, that repeated oral administration of L-ornithine at an early inactive period in mice induced a phase advance in the rhythm of PER2 expression. By contrast, L-ornithine administration to mouse embryonic fibroblasts did not affect the expression of PER2, indicating that L-ornithine indirectly alters the phase of PER2. L-Ornithine also increased plasma levels of insulin, glucose and glucagon-like peptide-1 alongside mPer2 expression, suggesting that it exerts its effects probably via insulin secretion. Collectively, these findings demonstrate that L-ornithine affects peripheral clock gene expression and may expand the possibilities of L-ornithine as a health food.

    DOI PubMed

  • Forced rather than voluntary exercise entrains peripheral clocks via a corticosterone/noradrenaline increase in PER2::LUC mice.

    Hiroyuki Sasaki, Yuta Hattori, Yuko Ikeda, Mayo Kamagata, Shiho Iwami, Shinnosuke Yasuda, Yu Tahara, Shigenobu Shibata

    Scientific reports   6   27607 - 27607  2016.06  [Refereed]  [International journal]

     View Summary

    Exercise during the inactive period can entrain locomotor activity and peripheral circadian clock rhythm in mice; however, mechanisms underlying this entrainment are yet to be elucidated. Here, we showed that the bioluminescence rhythm of peripheral clocks in PER2::LUC mice was strongly entrained by forced treadmill and forced wheel-running exercise rather than by voluntary wheel-running exercise at middle time during the inactivity period. Exercise-induced entrainment was accompanied by increased levels of serum corticosterone and norepinephrine in peripheral tissues, similar to the physical stress-induced response. Adrenalectomy with norepinephrine receptor blockers completely blocked the treadmill exercise-induced entrainment. The entrainment of the peripheral clock by exercise is independent of the suprachiasmatic nucleus clock, the main oscillator in mammals. The present results suggest that the response of forced exercise, but not voluntary exercise, may be similar to that of stress, and possesses the entrainment ability of peripheral clocks through the activation of the adrenal gland and the sympathetic nervous system.

    DOI PubMed

  • Anatomical cross-sectional area of the quadriceps femoris and sit-to-stand test score in middle-aged and elderly population: development of a predictive equation.

    Saito A, Ema R, Inami T, Maeo S, Otsuka S, Higuchi M, Shibata S, Kawakami Y

    Journal of physiological anthropology   36 ( 1 ) 3  2016.06  [Refereed]

    DOI PubMed

  • Anatomical cross-sectional area of the quadriceps femoris and sit-to-stand test score in middle-aged and elderly population: development of a predictive equation

    Akira Saito, Ryoichi Ema, Takayuki Inami, Sumiaki Maeo, Shun Otsuka, Mitsuru Higuchi, Shigenobu Shibata, Yasuo Kawakami

    Journal of Physiological Anthropology   36  2016.06  [Refereed]

     View Summary

    Background: Although the sit-to-stand (STS) test score has been shown to relate to the strength and size of the quadriceps femoris (QF) for elderly population, it is unknown whether this relationship is influenced by a posture (i.e., the trunk being allowed to stoop or not) during the STS test. The present study investigated the relationship between STS test score and QF anatomical cross-sectional area (ACSA) in the middle-aged and elderly population with regard to the difference in the posture during STS test, and aimed to develop an accurate predicting equation of the QF ACSA from the STS test score.
    Methods: 105 males (40-81 years) and 113 females (41-79 years) participated in the present study, then the subjects were divided at random as validation and cross-validation groups. Mid-thigh QF ACSA was determined by magnetic resonance imaging. Subjects performed a 10-repeated STS as fast as possible in two conditions: (1) with the trunk being allowed to stoop during the sitting phases, and (2) kept upright throughout the test. A power index of the STS test score was calculated based on an equation obtained in a previous study using the time taken for each test condition, the thigh and shank lengths, and body mass. In the validation group (n = 109), a stepwise multiple linear regression analysis was performed to create a predictive model of the ACSA with sex, age, the STS time, and power for both conditions as independent variables. The formulated predictive equation was examined in the cross-validation group (n = 109).
    Results: In the validation group, a stepwise regression analysis revealed that STS power with upright trunk condition, sex, and age but not with the stooping condition, were selected as variables to predict QF ACSA (R-2 = 0.64, P &lt; 0.001). There was no systematic error for the relationship between predicted and measured values in the cross-validation group.
    Conclusions: These results indicate that STS test score with upright trunk condition is one of the indices of QF muscle size of the middle-aged and elderly population. The estimated predicting equation should be useful in clinical and practical settings for the health promotion.

    DOI

  • Different Roles of Negative and Positive Components of the Circadian Clock in Oncogene-induced Neoplastic Transformation

    Chiharu Katamune, Satoru Koyanagi, Shoya Shiromizu, Naoya Matsunaga, Shigeki Shimba, Shigenobu Shibata, Shigehiro Ohdo

    JOURNAL OF BIOLOGICAL CHEMISTRY   291 ( 20 ) 10541 - +  2016.05  [Refereed]

     View Summary

    In mammals, circadian rhythms in physiological function are generated by a molecular oscillator driven by transcriptional-translational feedback loop consisting of negative and positive regulators. Disruption of this circadian clock machinery is thought to increase the risk of cancer development, but the potential contributions of each component of circadian clock to oncogenesis have been little explored. Here we reported that negative and positive transcriptional regulators of circadian feedback loop had different roles in oncogene-induced neoplastic transformation. Mouse embryonic fibroblasts prepared from animals deficient in negative circadian clock regulators, Period2 (Per2) or Cryptochrome1/2 (Cry1/2), were prone to transformation induced by co-expression of H-ras(V12) and SV40 large T antigen (SV40LT). In contrast, mouse embryonic fibroblasts prepared from mice deficient in positive circadian clock regulators, Bmal1 or Clock, showed resistance to oncogene-induced transformation. In Per2 mutant and Cry1/2-null cells, the introduction of oncogenes induced expression of ATF4, a potent repressor of cell senescence-associated proteins p16INK4a and p19ARF. Elevated levels of ATF4 were sufficient to suppress expression of these proteins and drive oncogenic transformation. Conversely, in Bmal1-null and Clock mutant cells, the expression of ATF4 was not induced by oncogene introduction, which allowed constitutive expression of p16INK4a and p19ARF triggering cellular senescence. Although genetic ablation of either negative or positive transcriptional regulators of the circadian clock leads to disrupted rhythms in physiological functions, our findings define their different contributions to neoplastic cellular transformation.

    DOI PubMed

  • Inhibition of IgE-mediated allergic reactions by pharmacologically targeting the circadian clock

    Yuki Nakamura, Nobuhiro Nakano, Kayoko Ishimaru, Noriko Ando, Ryohei Katoh, Katsue Suzuki-Inoue, Satoru Koyanagki, Hideoki Ogawa, Ko Okumura, Shigenobu Shibata, Atsuhito Nakao

    JOURNAL OF ALLERGY AND CLINICAL IMMUNOLOGY   137 ( 4 ) 1226 - 1235  2016.04  [Refereed]

     View Summary

    Background: The circadian clock temporally gates signaling through the high-affinity IgE receptor (Fc epsilon RI) in mast cells, thereby generating a marked day/night variation in allergic reactions. Thus manipulation of the molecular clock in mast cells might have therapeutic potential for IgE-mediated allergic reactions.
    Objective: We determined whether pharmacologically resetting the molecular clock in mast cells or basophils to times when FceRI signaling was reduced (ie, when core circadian protein period 2 [PER2] is upregulated) resulted in suppression of IgE-mediated allergic reactions.
    Methods: We examined the effects of PF670462, a selective inhibitor of the key clock component casein kinase 1 delta/epsilon, or glucocorticoid, both of which upregulated PER2 in mast cells, on IgE-mediated allergic reactions both in vitro and in vivo.
    Results: PF670462 or corticosterone (or dexamethasone) suppressed IgE-mediated allergic reactions in mouse bone marrow-derived mast cells or basophils and passive cutaneous anaphylactic reactions in mice in association with increased PER2 levels in mast cells or basophils. PF670462 or dexamethasone also ameliorated allergic symptoms in a mouse model of allergic rhinitis and downregulated allergen-specific basophil reactivity in patients with allergic rhinitis.
    Conclusion: Pharmacologically resetting the molecular clock in mast cells or basophils to times when Fc epsilon RI signaling is reduced can inhibit IgE-mediated allergic reactions. The results suggest a new strategy for controlling IgE-mediated allergic diseases. Additionally, this study suggests a novel mechanism underlying the antiallergic actions of glucocorticoids that relies on the circadian clock, which might provide a novel insight into the pharmacology of this drug in allergic patients.

    DOI PubMed

  • Leucine restores murine hepatic triglyceride accumulation induced by a low-protein diet by suppressing autophagy and excessive endoplasmic reticulum stress

    Shin-Ichi Yokota, Midori Ando, Shinya Aoyama, Kawai Nakamura, Shigenobu Shibata

    AMINO ACIDS   48 ( 4 ) 1013 - 1021  2016.04  [Refereed]

     View Summary

    Although it is known that a low-protein diet induces hepatic triglyceride (TG) accumulation in both rodents and humans, little is known about the underlying mechanism. In the present study, we modeled hepatic TG accumulation by inducing dietary protein deficiency in mice and aimed to determine whether certain amino acids could prevent low-protein diet-induced TG accumulation in the mouse liver. Mice fed a diet consisting of 3 % casein (3C diet) for 7 days showed hepatic TG accumulation with up-regulation of TG synthesis for the Acc gene and down-regulation of TG-rich lipoprotein secretion from hepatocytes for Mttp genes. Supplementing the 3 % casein diet with essential amino acids, branched-chain amino acids, or the single amino acid leucine rescued hepatic TG accumulation. In the livers of mice fed the 3 % casein diet, we observed a decrease in the levels of the autophagy substrate p62, an increase in the expression levels of the autophagy marker LC3-II, and an increase in the splicing of the endoplasmic reticulum (ER) stress-dependent Xbp1 gene. Leucine supplementation to the 3 % casein diet did not affect genes related to lipid metabolism, but inhibited the decrease in p62, the increase in LC3-II, and the increase in Xbp1 splicing levels in the liver. Our results suggest that ER stress responses and activated autophagy play critical roles in low-protein diet-induced hepatic TG accumulation in mice, and that leucine suppresses these two major protein degradation systems. This study contributes to understanding the mechanisms of hepatic disorders of lipid metabolism.

    DOI PubMed

  • Circadian rhythms of liver physiology and disease: experimental and clinical evidence.

    Yu Tahara, Shigenobu Shibata

    Nature reviews. Gastroenterology & hepatology   13 ( 4 ) 217 - 26  2016.04  [Refereed]  [International journal]

     View Summary

    The circadian clock system consists of a central clock located in the suprachiasmatic nucleus in the hypothalamus and peripheral clocks in peripheral tissues. Peripheral clocks in the liver have fundamental roles in maintaining liver homeostasis, including the regulation of energy metabolism and the expression of enzymes controlling the absorption and metabolism of xenobiotics. Over the past two decades, research has investigated the molecular mechanisms linking circadian clock genes with the regulation of hepatic physiological functions, using global clock-gene-knockout mice, or mice with liver-specific knockout of clock genes or clock-controlled genes. Clock dysfunction accelerates the development of liver diseases such as fatty liver diseases, cirrhosis, hepatitis and liver cancer, and these disorders also disrupt clock function. Food is an important regulator of circadian clocks in peripheral tissues. Thus, controlling the timing of food consumption and food composition, a concept known as chrononutrition, is one area of active research to aid recovery from many physiological dysfunctions. In this Review, we focus on the molecular mechanisms of hepatic circadian gene regulation and the relationships between hepatic circadian clock systems and liver physiology and disease. We concentrate on experimental data obtained from cell or mice and rat models and discuss how these findings translate into clinical research, and we highlight the latest developments in chrononutritional studies.

    DOI PubMed

  • Phase shifts in circadian peripheral clocks caused by exercise are dependent on the feeding schedule in PER2::LUC mice

    Hiroyuki Sasaki, Yuta Hattori, Yuko Ikeda, Mayo Kamagata, Shiho Iwami, Shinnosuke Yasuda, Shigenobu Shibata

    CHRONOBIOLOGY INTERNATIONAL   33 ( 7 ) 849 - 862  2016  [Refereed]

     View Summary

    Circadian rhythms are regulated by the suprachiasmatic nucleus (SCN) clock, which is the main oscillator and peripheral clock. SCN clock can be entrained by both photic and non-photic stimuli, and an interaction exists between photic and non-photic entrainment. Moreover, peripheral circadian clocks can be entrained not only by scheduled restricted feeding, but also by scheduled exercise. Thus, the entrainment of peripheral circadian clocks may be the result of an interaction between the entrainment caused by feeding and exercise. In this study, we examined the effect of wheel-running exercise on the phase of the peripheral clocks (kidney, liver and submandibular gland) in PER2::LUC mice under various feeding schedules. Phase and waveforms of the peripheral clocks were not affected by voluntary wheel-running exercise. Exercise for a period of 4 h during the early dark period (morning) delayed the peripheral clocks, while exercise for the same duration during the late dark period (evening) advanced the peripheral clocks. The feeding phase was advanced and delayed by evening and morning exercise, respectively, suggesting that the feeding pattern elicited by the scheduled exercise may entrain the peripheral clocks. Exercise did not affect the phase of the peripheral clock under the 1 meal per day schedule. When the phase of the peripheral clocks was advanced by the feeding schedule of 2 or 4 meals per day during light and/or dark periods, wheel-running exercise during the morning period significantly and equally shifted the phase of all organs back to the original positions observed in mice maintained under free-feeding conditions and with no exercise. When the schedule of 2 meals per day during the dark period failed to affect the phase of peripheral clock, morning exercise did not affect the phase. Wheel-running exercise increased the levels of serum corticosterone, and the injection of dexamethasone/corticosterone instead of exercise shifted a phase that had advanced under the feeding schedule of 2 meals per day, back to the normal position. The liver and submandibular glands exhibit higher sensitivity to dexamethasone than the kidneys. In adrenalectomized mice, treadmill-induced normalization of the advanced phase under a feeding schedule of 2 meals per day was not observed. In summary, scheduled exercise-induced phase shifts were weaker compared to scheduled feeding-induced phase shifts. The phase advance caused by the feeding schedule of 2 or 4 meals per day was suppressed by wheel-running, treadmill exercise or dexamethasone/corticosterone injection in the early dark period (morning). Corticosterone release may be involved in exercise-induced phase shift of peripheral clocks. These results suggest that there is an interaction between the phase shifts caused by feeding and exercise schedules in peripheral clocks.

    DOI PubMed

  • In vitro and in vivo phase changes of the mouse circadian clock by oxidative stress

    Yu Tahara, Aya Yokota, Takuya Shiraishi, Shunya Yamada, Atsushi Haraguchi, Ayako Shinozaki, Shigenobu Shibata

    Journal of Circadian Rhythms   14 ( 1 ) 1 - 7  2016  [Refereed]

     View Summary

    Mammalian circadian rhythms are governed by an endogenous circadian clock system, including the molecular clock works in each cell and tissue. Adaptation of the circadian clock to different environmental stimuli such as light, food, and stress is essential for homeostasis maintenance. However, the influence of oxidative stress on the circadian clock phase is not fully understood in vitro and in vivo. Here, we examined the effects of hydrogen peroxide (H2O2)-induced oxidative stress on the PERIOD2::LUCIFERASE bioluminescence rhythm in mouse embryonic fibroblasts in vitro and in mouse peripheral tissues in vivo. The circadian clock phase changed with the dose of H2O2 and time of day in vitro
    similar phase changes were observed in vivo in the circadian clocks of the peripheral tissues. In addition, mice treated with hemin-induced oxidative stress also showed phase changes of peripheral clocks, similarly as H2O2 treatment. Thus, oxidative stress can entrain circadian clock systems.

    DOI

  • Circadian Gene Clock Regulates Psoriasis-Like Skin Inflammation in Mice

    Noriko Ando, Yuki Nakamura, Rui Aoki, Kayoko Ishimaru, Hideoki Ogawa, Ko Okumura, Shigenobu Shibata, Shinji Shimada, Atsuhito Nakao

    JOURNAL OF INVESTIGATIVE DERMATOLOGY   135 ( 12 ) 3001 - 3008  2015.12  [Refereed]

     View Summary

    There are several reports suggesting that the pathophysiology of psoriasis may be associated with aberrant circadian rhythms. However, the mechanistic link between psoriasis and the circadian time-keeping system, "the circadian clock," remains unclear. This study determined whether the core circadian gene, Clock, had a regulatory role in the development of psoriasis. For this purpose, we compared the development of psoriasis-like skin inflammation induced by the Toll-like receptor 7 ligand imiquimod (IMQ) between wild-type mice and mice with a loss-of-function mutation of Clock. We also compared the development of IMQ-induced dermatitis between wild-type mice and mice with a loss-of-function mutation of Period2 (Per2), another key circadian gene that inhibits CLOCK activity. We found that Clock mutation ameliorated IMQ-induced dermatitis, whereas the Per2 mutation exaggerated IMQ-induced dermatitis, when compared with wild-type mice associated with decreased or increased IL-23 receptor (IL-23R) expression in gamma/delta(+) T cells, respectively. In addition, CLOCK directly bound to the promoter of IL-23R in gamma/delta(+) T cells, and IL-23R expression in the mouse skin was under circadian control. These findings suggest that Clock is a novel regulator of psoriasis-like skin inflammation in mice via direct modulation of IL-23R expression in gamma/delta(+) T cells, establishing a mechanistic link between psoriasis and the circadian clock.

    DOI PubMed

  • Impairment of Circadian Rhythms in Peripheral Clocks by Constant Light Is Partially Reversed by Scheduled Feeding or Exercise.

    Yutaro Hamaguchi, Yu Tahara, Masashi Hitosugi, Shigenobu Shibata

    Journal of biological rhythms   30 ( 6 ) 533 - 42  2015.12  [Refereed]  [International journal]

     View Summary

    In mammals, circadian rhythms in peripheral organs are impaired when animals are maintained in abnormal environmental light-dark cycles such as constant light (LL). This conclusion is based on averaged data from groups of experimental animals sacrificed at each time point. To investigate the effect of LL housing on the peripheral clocks of individual mice, an in vivo imaging system was used to observe the circadian bioluminescence rhythm in peripheral tissues of the liver, kidney, and submandibular salivary gland in PER2::LUCIFERASE knock-in mice. Using this technique, we demonstrated that the majority of individual peripheral tissues still had rhythmic oscillations of their circadian clocks in LL conditions. However, LL housing caused decreased amplitudes and a broad distribution of peak phases in PER2::LUCIFERASE oscillations irrespective of the state of the animals' behavioral rhythmicity. Because both scheduled feeding and scheduled exercise are effective recovery stimuli for circadian clock deficits, we examined whether scheduled feeding or scheduled exercise could reverse this impairment. The results showed that scheduled feeding or exercise could not restore the amplitude of peripheral clocks in LL. On the other hand, the LL-induced broad phase distribution was reversed, and peak phases were entrained to a specific time point by scheduled feeding but only slightly by scheduled exercise. The present results demonstrate that LL housing impairs peripheral circadian clock oscillations by altering both amplitude and phase in individual mice. The broad distribution of clock phases was clearly reversed by scheduled feeding, suggesting the importance of scheduled feeding as an entraining stimulus for impaired peripheral clocks.

    DOI PubMed

  • Phase-delay in the light-dark cycle impairs clock gene expression and levels of serotonin, norepinephrine, and their metabolites in the mouse hippocampus and amygdala.

    Shunpei Moriya, Yu Tahara, Hiroyuki Sasaki, Jun Ishigooka, Shigenobu Shibata

    Sleep medicine   16 ( 11 ) 1352 - 1359  2015.11  [Refereed]  [International journal]

     View Summary

    OBJECTIVE: A number of animal studies have implicated circadian clock genes in the regulation of mood, anxiety, and reward. However, the effect of misalignment of the environmental light-dark and internal circadian clock on the monoamine system is not fully understood. In the present study, we examined whether an abnormal light-dark schedule would affect behavior-, circadian clock-, and monoamine-related gene expressions, along with monoamine contents in the amygdala and hippocampus of mice. METHODS: Mice were subjected to an 8-hour phase delay in the light-dark cycle (Shift) every two days for four weeks, and locomotor activity was continuously measured. We examined the circadian expression of clock genes (Per1, Per2, and Bmal1) and genes of the NE/5HT uptake transporters (Net and Sert). In addition, the levels of NE/5HT and their metabolites MHPG/5HIAA were analyzed in the amygdala and hippocampus. RESULTS: Locomotor activity showed a free-running phenotype with a longer period (>24 hours) and showed misalignment between the light-dark and inactive-active cycles. The amplitude of the day-night fluctuation of Bmal1 expression was reduced in the amygdala and hippocampus of light-dark-shifted mice. Net gene expression in the Shift group showed different profiles compared with the Control group. In addition, NE and 5HT levels in the amygdala of the Shift group increased during the active period. CONCLUSIONS: The present results suggest that misalignment of the internal and external clocks by continuous shifting of the light-dark cycle affects the circadian clocks and monoamine metabolism in the amygdala and hippocampus of mice.

    DOI PubMed

  • Housing under abnormal light-dark cycles attenuates day/night expression rhythms of the clock genes Per1, Per2, and Bmal1 in the amygdala and hippocampus of mice.

    Shunpei Moriya, Yu Tahara, Hiroyuki Sasaki, Jun Ishigooka, Shigenobu Shibata

    Neuroscience research   99   16 - 21  2015.10  [Refereed]  [International journal]

     View Summary

    Although the results of previous studies have suggested that disruptions in circadian rhythms are involved in the pathogenesis of depression, no studies have examined the interaction of clock gene expression deficit and depression state. In this study, we examined clock gene expression levels and depressive-like behavior in mice housed under 3.5h light, 3.5h dark (T = 7) conditions to investigate the association between clock gene expression and depressive state. C57BL/6J mice were housed under a T = 24 cycle (12h light, 12h dark) or a T = 7 cycle and clock gene expression levels in the hippocampus and the amygdala were measured by real-time RT-PCR. Depressive state was evaluated by the forced swim test (FST). Although circadian rhythms of Per1 and Per2 clock gene expression in the hippocampus and amygdala were still detected under T = 7 conditions, rhythmicity and expression levels of both significantly decreased. Mice housed with a T = 7 cycle showed increased immobile time in the FST than those with a T = 24 cycle. The present results suggest that the presence of a depressive state around the early active phase of activity may be related to impairment of rhythmicity and expression levels of Per1 and Per2 genes under abnormal light-dark conditions.

    DOI PubMed

  • Artificial oxygen carriers rescue placental hypoxia and improve fetal development in the rat pre-eclampsia model

    Heng Li, Hidenobu Ohta, Yu Tahara, Sakiko Nakamura, Kazuaki Taguchi, Machiko Nakagawa, Yoshihisa Oishi, Yu-ichi Goto, Keiji Wada, Makiko Kaga, Masumi Inagaki, Masaki Otagiri, Hideo Yokota, Shigenobu Shibata, Hiromi Sakai, Kunihiro Okamura, Nobuo Yaegashi

    SCIENTIFIC REPORTS   5   15271  2015.10  [Refereed]

     View Summary

    Pre-eclampsia affects approximately 5% of all pregnant women and remains a major cause of maternal and fetal morbidity and mortality. The hypertension associated with pre-eclampsia develops during pregnancy and remits after delivery, suggesting that the placenta is the most likely origin of this disease. The pathophysiology involves insufficient trophoblast invasion, resulting in incomplete narrow placental spiral artery remodeling. Placental insufficiency, which limits the maternal-fetal exchange of gas and nutrients, leads to fetal intrauterine growth restriction. In this study, in our attempt to develop a new therapy for pre-eclampsia, we directly rescued placental and fetal hypoxia with nano-scale size artificial oxygen carriers (hemoglobin vesicles). The present study is the first to demonstrate that artificial oxygen carriers successfully treat placental hypoxia, decrease maternal plasma levels of anti-angiogenic proteins and ameliorate fetal growth restriction in the pre-eclampsia rat model.

    DOI PubMed

  • Effects of television luminance and wavelength at habitual bedtime on melatonin and cortisol secretion in humans

    Yoko Komada, Kazuyuki Aoki, Seiichi Gohshi, Hideki Ichioka, Shigenobu Shibata

    SLEEP AND BIOLOGICAL RHYTHMS   13 ( 4 ) 316 - 322  2015.10  [Refereed]

     View Summary

    The aim of this study was to examine the effect of exposure to different types of television displays at habitual bedtime on human melatonin and cortisol secretion. Thirteen male participants (mean age: 22.7 +/- 0.85 years) were tested over three nights in one baseline and two experimental sessions. Participants were instructed to watch a movie on four different luminance- and wavelength-controlled television displays: normal luminance (450 candela [cd]/m(2)) or high luminance (1200 cd/m(2)) and normal blue light or half blue light. Salivary melatonin and cortisol levels were measured at two time points before and after television viewing. There was no significant difference in cortisol secretion due to the different displays. Melatonin suppression was significantly lower following the exposure to the half-blue light display compared with the normal blue light display. These results suggest that the use of half-blue light displays during night time may prevent circadian rhythm dysfunction.

    DOI

  • 時計遺伝子Period2 非翻訳領域に結合する蛋白質の年齢依存的発現解析

    浜田俊幸, 宮川和子, 櫛笥博子, 柴田重信, 倉地須美子

    The Hokkaido Journal of Medical Sience   90 ( 2 )  2015.10  [Refereed]

  • Global Medical Discovery: Age-related expression analysis of mouse liver nuclear protein binding to 3'-untranslated region of Period2 gene

    HamadaT, Miyakawa K, Kushige H, Shibata S, Kurachi S

    Global Medical Discovery    2015.10  [Refereed]

    DOI

  • Antigen exposure in the late light period induces severe symptoms of food allergy in an OVA-allergic mouse model.

    Kana Tanabe, Eri Kitagawa, Misaki Wada, Atsushi Haraguchi, Kanami Orihara, Yu Tahara, Atsuhito Nakao, Shigenobu Shibata

    Scientific reports   5   14424 - 14424  2015.09  [Refereed]  [International journal]

     View Summary

    The mammalian circadian clock controls many physiological processes that include immune responses and allergic reactions. Several studies have investigated the circadian regulation of intestinal permeability and tight junctions known to be affected by cytokines. However, the contribution of circadian clock to food allergy symptoms remains unclear. Therefore, we investigated the role of the circadian clock in determining the severity of food allergies. We prepared an ovalbumin food allergy mouse model, and orally administered ovalbumin either late in the light or late in the dark period under light-dark cycle. The light period group showed higher allergic diarrhea and weight loss than the dark period group. The production of type 2 cytokines, IL-13 and IL-5, from the mesenteric lymph nodes and ovalbumin absorption was higher in the light period group than in the dark period group. Compared to the dark period group, the mRNA expression levels of the tight junction proteins were lower in the light period group. We have demonstrated that increased production of type 2 cytokines and intestinal permeability in the light period induced severe food allergy symptoms. Our results suggest that the time of food antigen intake might affect the determination of the severity of food allergy symptoms.

    DOI PubMed

  • Entrainment of mouse peripheral circadian clocks to <24 h feeding/fasting cycles under 24 h light/dark conditions.

    Yutaro Hamaguchi, Yu Tahara, Hiroaki Kuroda, Atsushi Haraguchi, Shigenobu Shibata

    Scientific reports   5   14207 - 14207  2015.09  [Refereed]  [International journal]

     View Summary

    The circadian clock system in peripheral tissues can endogenously oscillate and is entrained by the light-dark and fasting-feeding cycles in mammals. Although the system's range of entrainment to light-dark cycles with a non-24 h (<24 h) interval has been studied, the range of entrainment to fasting-feeding cycles with shorter periods (<24 h) has not been investigated in peripheral molecular clocks. In the present study, we measured this range by monitoring the mouse peripheral PER2::LUCIFERASE rhythm in vivo at different periods under each feeding cycle (Tau (T) = 15-24 h) under normal light-dark conditions. Peripheral clocks could be entrained to the feeding cycle with T = 22-24 h, but not to that with T = 15-21 h. Under the feeding cycle with T = 15-18 h, the peripheral clocks oscillated at near the 24-h period, suggesting that they were entrained to the light-dark cycle. Thus, for the first time, we demonstrated the range of entrainment to the non-24 h feeding cycle, and that the circadian range (T = 22-24 h) of feeding stimulus is necessary for peripheral molecular clock entrainment under light-dark cycles.

    DOI PubMed

  • Age-related expression analysis of mouse liver nuclear protein binding to 3'-untranslated region of Period2 gene

    Toshiyuki Hamada, Kazuko Miyakawa, Hiroko Kushige, Shigenobu Shibata, Sumiko Kurachi

    JOURNAL OF PHYSIOLOGICAL SCIENCES   65 ( 4 ) 349 - 357  2015.07  [Refereed]

     View Summary

    In mammals, both circadian rhythm and aging play important roles in regulating time-dependent homeostasis. We previously discovered an age-related increase element binding protein, hnRNP A3, which binds to the 3'-untranslated region (UTR) of blood coagulation factor IX (FIX). Here, we describe other members of this protein family, hnRNP C and hnRNP H, which bind to the 3'-UTR of the mouse circadian clock gene Period 2 (mPer2). RNA electrophoretic mobility shift assays using a P-32-labeled Per2 RNA probe coupled with two-dimensional gel electrophoresis followed by MALDI-TOF/MS peptide mass fingerprint analysis was used to analyze these proteins. Western blotting suggested that the total expression of these proteins in mouse liver cell nuclei does not increase with age. Two-dimensional gel electrophoresis analysis of age-related protein expression showed that many isoforms of these proteins exist in the liver and that each protein exhibits a complex age-related expression pattern. These results suggest that many isoforms of proteins are regulated by different aging systems and that many age regulation systems function in the liver.

    DOI PubMed

  • Entrainment of the mouse circadian clock by sub-acute physical and psychological stress.

    Yu Tahara, Takuya Shiraishi, Yosuke Kikuchi, Atsushi Haraguchi, Daisuke Kuriki, Hiroyuki Sasaki, Hiroaki Motohashi, Tomoko Sakai, Shigenobu Shibata

    Scientific reports   5   11417 - 11417  2015.06  [Refereed]  [International journal]

     View Summary

    The effects of acute stress on the peripheral circadian system are not well understood in vivo. Here, we show that sub-acute stress caused by restraint or social defeat potently altered clock gene expression in the peripheral tissues of mice. In these peripheral tissues, as well as the hippocampus and cortex, stressful stimuli induced time-of-day-dependent phase-advances or -delays in rhythmic clock gene expression patterns; however, such changes were not observed in the suprachiasmatic nucleus, i.e. the central circadian clock. Moreover, several days of stress exposure at the beginning of the light period abolished circadian oscillations and caused internal desynchronisation of peripheral clocks. Stress-induced changes in circadian rhythmicity showed habituation and disappeared with long-term exposure to repeated stress. These findings suggest that sub-acute physical/psychological stress potently entrains peripheral clocks and causes transient dysregulation of circadian clocks in vivo.

    DOI PubMed

  • Eating meals before wheel-running exercise attenuate high fat diet-driven obesity in mice under two meals per day schedule

    Hiroyuki Sasaki, Yuta Hattori, Yuko Ikeda, Mayo Kamagata, Shigenobu Shibata

    CHRONOBIOLOGY INTERNATIONAL   32 ( 5 ) 677 - 686  2015.06  [Refereed]

     View Summary

    Mice that exercise after meals gain less body weight and visceral fat compared to those that exercised before meals under a one meal/exercise time per day schedule. Humans generally eat two or three meals per day, and rarely have only one meal. To extend our previous observations, we examined here whether a "two meals, two exercise sessions per day'' schedule was optimal in terms of maintaining a healthy body weight. In this experiment, "morning'' refers to the beginning of the active phase (the "morning'' for nocturnal animals). We found that 2-h feeding before 2-h exercise in the morning and evening (F-Ex/F-Ex) resulted in greater attenuation of high fat diet (HFD)-induced weight gain compared to other combinations of feeding and exercise under two daily meals and two daily exercise periods. There were no significant differences in total food intake and total wheel counts, but feeding before exercise in the morning groups (F-Ex/F-Ex and F-Ex/Ex-F) increased the morning wheel counts. These results suggest that habitual exercise after feeding in the morning and evening is more effective for preventing HFD-induced weight gain. We also determined whether there were any correlations between food intake, wheel rotation, visceral fat volume and skeletal muscle volumes. We found positive associations between gastrocnemius muscle volumes and morning wheel counts, as well as negative associations between morning food intake volumes/body weight and morning wheel counts. These results suggest that morning exercise-induced increase of muscle volume may refer to anti-obesity. Evening exercise is negatively associated with fat volume increases, suggesting that this practice may counteract fat deposition. Our multifactorial analysis revealed that morning food intake helps to increase exercise, and that evening exercise reduced fat volumes. Thus, exercise in the morning or evening is important for preventing the onset of obesity.

    DOI PubMed

  • 毒性学における生体リズムの重要性を考える 中枢神経系作用薬の時間薬理学とニューロン新生

    守屋 孝洋, 竹生田 淳, 茂木 明日香, 佐々木 崇志, 前川 知子, 鈴木 登紀子, 柴田 重信, 太田 英伸, 小林 正樹

    The Journal of Toxicological Sciences   40 ( Suppl. ) S55 - S55  2015.06

  • Nutrition and Diet as Potent Regulators of the Liver Clock

    Yu Tahara, Shigenobu Shibata

    Circadian Medicine     107 - 117  2015.05  [Refereed]

     View Summary

    This chapter centers on the new keyword "chrono-nutrition", which helps to understand the importance of mutual interactions between circadian rhythms and nutrition/diet. The timing of food can have a major impact on the circadian system. The "food entrainable oscillator" (FEO) senses food timing and/or nutritional factors, and then organizes the circadian system independent of suprachiasmatic nucleus (SCN) regulation. To understand the mechanism of FEO, many types of food have been used for food-induced phase entrainment in the circadian system. Food-induced insulin secretion is an important direct pathway for food entrainment in the liver circadian clock. The chapter also highlights the interactions between food, circadian system, and disease. Regular meal time has the possibility to improve the deficit of the circadian system, independent of the master clock function. Food can become a "zeitgeber" to entrain the circadian system and it is hoped that eating times function as a "zeitgeber" in humans.

    DOI

  • The Circadian Clock Functions As A Potent Regulator of Allergic Reaction. [Review]

    Atsuhito NAKAO, Yuki NAKAMURA, Shibata S

    Allergy   70 ( 5 ) 467 - 473  2015.05  [Refereed]

     View Summary

    IF:6.335

  • Feeding and adrenal entrainment stimuli are both necessary for normal circadian oscillation of peripheral clocks in mice housed under different photoperiods.

    Yuko Ikeda, Hiroyuki Sasaki, Teiji Ohtsu, Takuya Shiraishi, Yu Tahara, Shigenobu Shibata

    Chronobiology international   32 ( 2 ) 195 - 210  2015.03  [Refereed]  [International journal]

     View Summary

    The mammalian circadian rhythm is entrained by multiple factors, including the light-dark cycle, the organism's feeding pattern and endocrine hormones such as glucocorticoids. Both a central clock (the suprachiasmatic nucleus, or SCN) and peripheral clocks (i.e. in the liver and lungs) in mice are entrained by photoperiod. However, the factors underlying entrainment signals from the SCN to peripheral clocks are not well known. To elucidate the role of entrainment factors such as corticosterone and feeding, we examined whether peripheral clock rhythms were impaired by adrenalectomy (ADX) and/or feeding of 6 meals per day at equal intervals under short-day, medium-day and long-day photoperiods (SP, MP and LP, respectively). We evaluated the waveform and phase of circadian rhythms in the liver, kidney and salivary gland by in vivo imaging of PER2::LUCIFERASE knock-in mice. In intact mice, the waveforms of the peripheral clocks were similar among all photoperiods. The phases of peripheral clocks were well adjusted by the timing of the "lights-off"-operated evening (E) oscillator but not the "lights-on"-operated morning (M) oscillator. ADX had almost no effect on the rhythmicity and phase of peripheral clocks, regardless of photoperiod. To reduce the feeding-induced signal, we placed mice on a restricted feeding regimen with 6 meals per day (6 meals RF). This caused advances of the peripheral clock phase in LP-housed mice (2-5 h) and MP-housed mice (1-2 h) but not SP-housed mice. Thus, feeding pattern may affect the phase of peripheral clocks, depending on photoperiod. More specifically, ADX + 6 meals RF mice showed impairment of circadian rhythms in the kidney and liver but not in the salivary gland, regardless of photoperiod. However, the impairment of peripheral clocks observed in ADX + 6 meals RF mice was reversed by administration of dexamethasone for 3 days. The phase differences in the salivary gland clock among SP-, MP- and LP-housed mice became very small following treatment with ADX + 6 meals RF, suggesting that the effect of photoperiod was reduced by ADX and 6 meals RF. Because the SCN rhythm (as evaluated by PER2 immunohistochemistry) was not disrupted by ADX + 6 meals RF, impairment of peripheral clocks in these mice was not because of impaired SCN clock function. In addition, locomotor activity rhythm and modifications of the feeding pattern may not be completely responsible for determining the phase of peripheral clocks. Thus, this study demonstrates that the phase of peripheral clocks responds to a photoperiodic lights-off signal, and suggests that signals from normal feeding patterns and the adrenal gland are necessary to maintain the oscillation and phase of peripheral clocks under various photoperiods.

    DOI PubMed

  • Fish Oil Accelerates Diet-Induced Entrainment of the Mouse Peripheral Clock via GPR120.

    Akiko Furutani, Yuko Ikeda, Misa Itokawa, Hiroki Nagahama, Teiji Ohtsu, Naoki Furutani, Mayo Kamagata, Zhi-Hong Yang, Akira Hirasawa, Yu Tahara, Shigenobu Shibata

    PloS one   10 ( 7 ) e0132472  2015  [Refereed]  [International journal]

     View Summary

    The circadian peripheral clock is entrained by restricted feeding (RF) at a fixed time of day, and insulin secretion regulates RF-induced entrainment of the peripheral clock in mice. Thus, carbohydrate-rich food may be ideal for facilitating RF-induced entrainment, although the role of dietary oils in insulin secretion and RF-induced entrainment has not been described. The soybean oil component of standard mouse chow was substituted with fish or soybean oil containing docosahexaenoic acid (DHA) and/or eicosapentaenoic acid (EPA). Tuna oil (high DHA/EPA), menhaden oil (standard), and DHA/EPA dissolved in soybean oil increased insulin secretion and facilitated RF-induced phase shifts of the liver clock as represented by the bioluminescence rhythms of PER2::LUCIFERASE knock-in mice. In this model, insulin depletion blocked the effect of tuna oil and fish oil had no effect on mice deficient for GPR120, a polyunsaturated fatty acid receptor. These results suggest food containing fish oil or DHA/EPA is ideal for adjusting the peripheral clock.

    DOI PubMed

  • The circadian clock controls fluctuations of colonic cell proliferation during the light/dark cycle via feeding behavior in mice

    Daisuke Yoshida, Natsumi Aoki, Mizuho Tanaka, Shinya Aoyama, Shigenobu Shibata

    CHRONOBIOLOGY INTERNATIONAL   32 ( 8 ) 1145 - 1155  2015  [Refereed]

     View Summary

    The mammalian circadian system is controlled not only by the suprachiasmatic nucleus (SCN), but also by the peripheral clocks located in tissues such as liver, kidney, small intestine, and colon, mediated through signals such as hormones. Peripheral clocks, but not the SCN, can be entrained by food intake schedules. While it is known that cell proliferation exhibits a circadian rhythm in the colon epithelium, it is unclear how this rhythm is influenced by food intake schedules. Here, we aimed to determine the relationships between feeding schedules and cell proliferation in the colon epithelium by means of immunochemical analysis, using 5-bromo-2'-deoxyuridine (BrdU), as well as to elucidate how feeding schedules influence the colonic expression of clock and cell cycle genes, using real-time reverse-transcription PCR (qRT-PCR). Cell proliferation in the colonic epithelium of normal mice exhibited a daily fluctuation, which was abrogated in Clock mutant mice. The day/night pattern of cellular proliferation and clock gene expression under daytime and nighttime restricted feeding (RF) schedules showed opposite tendencies. While daytime RF for every 4 h attenuated the day/night pattern of cell proliferation, this was restored to normal in the Clock mutant mice under the nighttime RF schedule. These results suggest that feeding schedules contribute to the establishment of a daily fluctuation of cell proliferation and RF can recover it in Clock mutant mice. Thus, this study demonstrates that the daily fluctuation of cell proliferation in the murine colon is controlled by a circadian feeding rhythm, suggesting that feeding schedules are important for rhythmicity in the proliferation of colon cells.

    DOI PubMed

  • Circadian rhythm and food/nutrition

    Yu Tahara, Shigenobu Shibata

    Mechanisms of Circadian Systems in Animals and Their Clinical Relevance     237 - 259  2015.01  [Refereed]

     View Summary

    Scheduled food access during the daytime for nocturnal mice or rats entrains the food-entrainable oscillator (FEO) in the brain and the food-entrainable peripheral oscillator (FEPO) in the peripheral tissues. FEO and FEPO are not regulated by the central clock, which is in the suprachiasmatic nucleus. FEO produces food anticipatory activity (FAA) 2-3 h before the scheduled feeding time initiates. FEPO produces entrainment in the rhythm of peripheral clock gene expression and in the rhythm of food-metabolic functions in peripheral organs. At present, the mechanisms of the FEO and FEPO are not completely understood, despite many studies that have been performed in this field. In addition, circadian clocks affect metabolism of nutrition (absorption, distribution, metabolism, excretion). In this review, we describe and review the characteristics and biological implications of FEO and FEPO and the mechanism of metabolism of nutrition with day-night differences. We call this relationship between nutrition and chronobiology –Chrono-nutrition,— which is an important study field to understand how our body clocks contribute to our health throughout the day.

    DOI

  • Effects of caffeine on circadian phase, amplitude and period evaluated in cells in vitro and peripheral organs in vivo in PER2::LUCIFERASE mice.

    Seira Narishige, Mari Kuwahara, Ayako Shinozaki, Satoshi Okada, Yuko Ikeda, Mayo Kamagata, Yu Tahara, Shigenobu Shibata

    British journal of pharmacology   171 ( 24 ) 5858 - 69  2014.12  [Refereed]  [International journal]

     View Summary

    BACKGROUND AND PURPOSE: Caffeine is one of the most commonly used psychoactive substances. Circadian rhythms consist of the main suprachiasmatic nucleus (SCN) clocks and peripheral clocks. Although caffeine lengthens circadian rhythms and modifies phase changes in SCN-operated rhythms, the effects on caffeine on the phase, period and amplitude of peripheral organ clocks are not known. In addition, the role of cAMP/Ca(2+) signalling in effects of caffeine on rhythm has not been fully elucidated. EXPERIMENTAL APPROACH: We examined whether chronic or transient application of caffeine affects circadian period/amplitude and phase by evaluating bioluminescence rhythm in PER2::LUCIFERASE knock-in mice. Circadian rhythms were monitored in vitro using fibroblasts and ex vivo and in vivo for monitoring of peripheral clocks. KEY RESULTS: Chronic application of caffeine (0.1-10 mM) increased period and amplitude in vitro. Transient application of caffeine (10 mM) near the bottom of the decreasing phase of bioluminescence rhythm caused phase advance in vitro. Caffeine (0.1%) intake caused a phase delay under light-dark or constant dark conditions, suggesting a period-lengthening effect in vivo. Caffeine (20 mg·kg(-1) ) at daytime or at late night-time caused phase advance or delay in bioluminescence rhythm in the liver and kidney respectively. The complicated roles of cAMP/Ca(2+) signalling may be involved in the caffeine-induced increase of period and amplitude in vitro. CONCLUSIONS AND IMPLICATIONS: Caffeine affects circadian rhythm in mice by lengthening the period and causing a phase shift of peripheral clocks. These results suggest that caffeine intake with food/drink may help with food-induced resetting of peripheral circadian clocks.

    DOI PubMed

  • Combination of meal and exercise timing with a high-fat diet influences energy expenditure and obesity in mice

    Hiroyuki Sasaki, Teiji Ohtsu, Yuko Ikeda, Miku Tsubosaka, Shigenobu Shibata

    CHRONOBIOLOGY INTERNATIONAL   31 ( 9 ) 959 - 975  2014.11  [Refereed]

     View Summary

    In mice, obesity has been observed not only in those freely fed a high-fat diet (HFD) but also in those fed while physically inactive. In contrast, a HFD during physically active periods protects against obesity and the impairments in the circadian rhythm induced by free feeding of a HFD. Although exercise is known to be effective for obesity prevention and management, the optimal timing of exercise has not yet been determined. In the present experiments, we aimed to determine the best combination of daily timing of HFD consumption and exercise for the prevention of HFD-induced weight gain in mice. In this experiment, "morning" refers to the beginning of the active phase (the "morning" for nocturnal animals). Increases in body weight related to free feeding of a HFD was significantly reduced with 4 h of exercise during the late (evening) or middle (noon) active period compared to 4 h of exercise during the early (morning) active period or free access to exercise, which resulted in hours of exercise similar to that of morning exercise. These results suggested that eating in the morning or at noon followed by exercise in the evening could prevent weight gain more effectively than exercise in the morning followed by eating at noon or in the evening. The group fed a HFD for 4 h in the morning had lower body weight than the group fed a HFD for 4 h in the evening without exercise. The last group of experiments tested the hypothesis that there would be an interaction between mealtime and exercise time (i.e. time of day) versus order (i.e. which comes first) effects. We compared groups that exercised for 4 h at noon and were fed either in the morning or evening and groups that were fed for 4 h at noon and either exercised in the morning or evening. We found that the groups that were fed before exercise gained less body and fat weight and more skeletal muscle weight compared to the groups that exercised before eating. Corresponding to the body and fat weight changes, the respiratory exchange ratio (RER) was lower and energy expenditure was higher in the groups fed before exercise than in the groups fed after exercise, and these effects on energy metabolism were also observed in the early stage of HFD feeding before obesity. When obese mice fed a HFD for 12 weeks were exposed to a combination of feeding and exercise timing in an effort to reduce body weight, eating followed by exercise resulted in greater weight loss, similar to the experiments conducted to prevent weight gain. These results demonstrate that a combination of daily timing of eating and exercise may influence weight gain and that eating followed by exercise may be effective for minimizing increases in body and fat weight as well as maximizing increases in skeletal muscle weight.

    DOI PubMed

  • Controlling access time to a high-fat diet during the inactive period protects against obesity in mice.

    Atsushi Haraguchi, Natsumi Aoki, Teiji Ohtsu, Yuko Ikeda, Yu Tahara, Shigenobu Shibata

    Chronobiology international   31 ( 8 ) 935 - 44  2014.10  [Refereed]  [International journal]

     View Summary

    Free feeding (FF) with a high fat diet (HFD) causes excessive body weight gain, whereas restricted feeding (RF) with a HFD attenuates body weight gain. The effects of timing of feeding with a HFD (day vs. night) and feeding duration on energy homeostasis have not yet been investigated. In this study, we fed mice a HFD or a normal diet (ND) twice a day, during their active and inactive periods, on a schedule. The amount of food was regulated by feeding duration (2, 4 or 8 h). First, we investigated the effects of 4-h RF during active-inactive periods (ND-ND, HFD-HFD, ND-HFD or HFD-ND). Among all the 4-h RF groups, mice consumed almost the same amount of calories as those in the FF[ND] group, even those fed a HFD. Body weight and visceral fat in these three groups were lower than that in the FF[HFD] group. Second, we investigated the effects of RF duration. Body weight and visceral fat were higher in the 8-h groups than in the 4-h groups. Body weight and visceral fat were higher in the 2-h groups than in the 4-h groups even though the 2-h groups had less food. Third, we investigated the effects of eating a HFD during the inactive period, when RF duration was extended (2, 6 or 12 h). Mice were fed with a HFD during the inactive period for 2 h and fed with a ND during the active period for 2, 6 or 12 h. Body weight and visceral fat in these mice were comparable to those in the FF[ND] mice. The results of our first set of experiments suggest that 4-h RF was an adequate feeding duration to control the effect of a HFD on obesity. The results of our second set of experiments suggest 2-h RF (such as speed-eating) and 8-h RF, representative of eating disorders, are unhealthy feeding patterns related to obesity. The results of our third set of experiments suggest that eating a HFD for a short period during the night does not affect body weight and visceral fat. Taken together, these results indicate that consideration to feeding with a HFD during the inactive period and restricting eating habits relieve the risks of body weight gain and visceral fat accumulation.

    DOI PubMed

  • Bile Acid-regulated Peroxisome Proliferator-activated Receptor-alpha (PPAR alpha) Activity Underlies Circadian Expression of Intestinal Peptide Absorption Transporter PepT1/Slc15a1

    Ayako Okamura, Satoru Koyanagi, Adila Dilxiat, Naoki Kusunose, Jia Jun Chen, Naoya Matsunaga, Shigenobu Shibata, Shigehiro Ohdo

    JOURNAL OF BIOLOGICAL CHEMISTRY   289 ( 36 ) 25296 - 25305  2014.09  [Refereed]

     View Summary

    Digested proteins are mainly absorbed as small peptides composed of two or three amino acids. The intestinal absorption of small peptides is mediated via only one transport system: the proton-coupled peptide transporter-1 (PepT1) encoded from the soluble carrier protein Slc15a1. In mammals, intestinal expression of PepT1/Slc15a1 oscillates during the daily feeding cycle. Although the oscillation in the intestinal expression of PepT1/Slc15a1 is suggested to be controlled by molecular components of circadian clock, we demonstrated here that bile acids regulated the oscillation of PepT1/Slc15a1 expression through modulating the activity of peroxisome proliferator-activated receptor alpha (PPAR alpha). Nocturnally active mice mainly consumed their food during the dark phase. PPAR alpha activated the intestinal expression of Slc15a1 mRNA during the light period, and protein levels of PepT1 peaked before the start of the dark phase. After food intake, bile acids accumulated in intestinal epithelial cells. Intestinal accumulated bile acids interfered with recruitment of co-transcriptional activator CREB-binding protein/p300 on the promoter region of Slc15a1 gene, thereby suppressing PPAR alpha-mediated transactivation of Slc15a1. The time-dependent suppression of PPAR alpha-mediated transactivation by bile acids caused an oscillation in the intestinal expression of PepT1/Slc15a1 during the daily feeding cycle that led to circadian changes in the intestinal absorption of small peptides. These findings suggest a molecular clock-independent mechanism by which bile acid-regulated PPAR alpha activity governs the circadian expression of intestinal peptide transporter.

    DOI PubMed

  • A single daily meal at the beginning of the active or inactive period inhibits food deprivation-induced fatty liver in mice.

    Natsumi Aoki, Daisuke Yoshida, Ryosuke Ishikawa, Midori Ando, Kaai Nakamura, Yu Tahara, Shigenobu Shibata

    Nutrition research (New York, N.Y.)   34 ( 7 ) 613 - 22  2014.07  [Refereed]  [International journal]

     View Summary

    Food deprivation (FD) induces hepatic steatosis in both rodents and humans. Although body composition, age, and sex influence hepatic triglyceride (TG) levels after FD, whether feeding patterns affect FD-induced liver TG increases is unknown. We hypothesized that restricted feeding (RF) of 1 meal per day during the active or inactive period (especially the inactive period) augments FD-induced elevation of liver TGs because RF in the inactive period impairs the circadian rhythm. Triglyceride levels and the expression of genes related to TG metabolism in the liver were examined by a bioassay and real-time reverse transcription-polymerase chain reaction, respectively. In the first experiment, when compared to nonfasted mice, mice that fasted for 24 hours showed a 1.5-fold (FD starting during the inactive period) to 3-fold (FD started during the active period) increase in liver TG levels. This experiment showed that TG levels depend upon the starting time of FD. In the second experiment, mice were given free access to food for 3 hours at the beginning of either the inactive ("supper-only") or the active ("breakfast-only") period for 2 weeks. Restricted feeding inhibited the FD-induced increases in liver and serum TG levels, serum free fatty acids, and the expression of genes related to fatty acid uptake in the liver, including fatty acid transport protein 1 (Fatp1) and 4 (Fatp4). Unexpectedly, compared to free feeding, RF during the active or inactive period resulted in resistance to FD-induced fatty liver. This is the first study to demonstrate that feeding patterns affect FD-induced TG accumulation in the mouse liver.

    DOI PubMed

  • Expressions of Tight Junction Proteins Occludin and Claudin-1 Are under the Circadian Control in the Mouse Large Intestine: Implications in Intestinal Permeability and Susceptibility to Colitis

    Kyoko Oh-oka, Hiroshi Kono, Kayoko Ishimaru, Kunio Miyake, Takeo Kubota, Hideoki Ogawa, Ko Okumura, Shigenobu Shibata, Atsuhito Nakao

    PLOS ONE   9 ( 5 ) e98016  2014.05  [Refereed]

     View Summary

    Background & Aims: The circadian clock drives daily rhythms in behavior and physiology. A recent study suggests that intestinal permeability is also under control of the circadian clock. However, the precise mechanisms remain largely unknown. Because intestinal permeability depends on tight junction (TJ) that regulates the epithelial paracellular pathway, this study investigated whether the circadian clock regulates the expression levels of TJ proteins in the intestine.
    Methods: The expression levels of TJ proteins in the large intestinal epithelium and colonic permeability were analyzed every 4, 6, or 12 hours between wild-type mice and mice with a mutation of a key clock gene Period2 (Per2; mPer2(m/m)). In addition, the susceptibility to dextran sodium sulfate (DSS)-induced colitis was compared between wild-type mice and mPer2(m/m) mice.
    Results: The mRNA and protein expression levels of Occludin and Claudin-1 exhibited daily variations in the colonic epithelium in wild-type mice, whereas they were constitutively high in mPer2(m/m) mice. Colonic permeability in wild-type mice exhibited daily variations, which was inversely associated with the expression levels of Occludin and Claudin-1 proteins, whereas it was constitutively low in mPer2(m/m) mice. mPer2(m/m) mice were more resistant to the colonic injury induced by DSS than wild-type mice.
    Conclusions: Occludin and Claudin-1 expressions in the large intestine are under the circadian control, which is associated with temporal regulation of colonic permeability and also susceptibility to colitis.

    DOI PubMed

  • Circadian regulation of allergic reactions by the mast cell clock in mice.

    Yuki Nakamura, Nobuhiro Nakano, Kayoko Ishimaru, Mutsuko Hara, Takako Ikegami, Yu Tahara, Ryohei Katoh, Hideoki Ogawa, Ko Okumura, Shigenobu Shibata, Chiharu Nishiyama, Atsuhito Nakao

    The Journal of allergy and clinical immunology   133 ( 2 ) 568 - 75  2014.02  [Refereed]  [International journal]

     View Summary

    BACKGROUND: It remains elusive how allergic symptoms exhibit prominent 24-hour variations. In mammals the circadian clocks present in nearly all cells, including mast cells, drive the daily rhythms of physiology. Recently, we have shown that the circadian clocks drive the daily rhythms in IgE/mast cell-mediated allergic reactions. However, the precise mechanisms, particularly the specific roles of the mast cell-intrinsic clockwork in temporal regulation, remain unclear. OBJECTIVE: We determined whether the mast cell clockwork contributes to the temporal regulation of IgE/mast cell-mediated allergic reaction. METHODS: The kinetics of a time of day-dependent variation in passive cutaneous anaphylactic reactions were compared between mast cell-deficient mice reconstituted with bone marrow-derived cultured mast cells generated from mice with a wild-type allele and a dominant negative type mutation of the key clock gene Clock. We also examined the temporal responses of wild-type and Clock-mutated bone marrow-derived cultured mast cells to IgE stimulation in vitro. Furthermore, factors influencing the mast cell clockwork were determined by using in vivo imaging. RESULTS: The Clock mutation in mast cells resulted in the absence of temporal variations in IgE-mediated degranulation in mast cells both in vivo and in vitro associated with the loss of temporal regulation of FcεRI expression and signaling. Additionally, adrenalectomy abolished the mast cell clockwork in vivo. CONCLUSION: The mast cell-intrinsic clockwork, entrained by humoral factors from the adrenal gland, primarily contributes to the temporal regulation of IgE/mast cell-mediated allergic reactions. Our results reveal a novel regulatory mechanism for IgE-mediated mast cell responses that might underlie the circadian pathophysiology in patients with allergic diseases.

    DOI PubMed

  • Involvement of 5-HT3 and 5-HT4 Receptors in the Regulation of Circadian Clock Gene Expression in Mouse Small Intestine

    Natsumi Aoki, Hiroyuki Watanabe, Kazuya Okada, Kazuyuki Aoki, Takuma Imanishi, Daisuke Yoshida, Ryosuke Ishikawa, Shigenobu Shibata

    JOURNAL OF PHARMACOLOGICAL SCIENCES   124 ( 2 ) 267 - 275  2014.02  [Refereed]

     View Summary

    Several lines of evidence suggest that 5-HT receptors play a critical role in the expression of clock genes in the suprachiasmatic nucleus, the main circadian oscillator in hamsters. The contributions of 5-HT-receptor subtypes in the intestine, where they are expressed at high concentrations, are however not yet clarified. The 5-HT synthesis inhibitor,p-chlorophenylalanine, attenuated the daily rhythm of Per1 and Per2 gene expression in the intestine. Injection of 5-HT and agonists of the 5-HT3 and 5-HT4 receptors increased Per1/Per2 expression and decreased Bmall expression in a dose-dependent manner Although treatment with antagonists of 5-HT3 and 5-HT4 alone did not affect clock gene expression, co-injection of these antagonists with 5-HT blocked the 5-HT-induced changes in clock gene expression. Increased tissue levels of 5-HT due to treatment with the antidepressants clomipramine and fluvoxamine did not affect clock gene expression. The present results suggest that the 5-HT system in the small intestine may play a critical role in regulating circadian rhythms through 5-HT3/5-HT4-receptor activation.

    DOI PubMed

  • Chrono-biology, chrono-pharmacology, and chrono-nutrition.

    Yu Tahara, Shigenobu Shibata

    Journal of pharmacological sciences   124 ( 3 ) 320 - 35  2014  [Refereed]  [Domestic journal]

     View Summary

    The circadian clock system in mammals drives many physiological processes including the daily rhythms of sleep-wake behavior, hormonal secretion, and metabolism. This system responds to daily environmental changes, such as the light-dark cycle, food intake, and drug administration. In this review, we focus on the central and peripheral circadian clock systems in response to drugs, food, and nutrition. We also discuss the adaptation and anticipation mechanisms of our body with regard to clock system regulation of various kinetic and dynamic pathways, including absorption, distribution, metabolism, and excretion of drugs and nutrients. "Chrono-pharmacology" and "chrono-nutrition" are likely to become important research fields in chrono-biological studies.

    DOI PubMed

  • Disruption of the suprachiasmatic nucleus blunts a time of day-dependent variation in systemic anaphylactic reaction in mice.

    Yuki Nakamura, Kayoko Ishimaru, Yu Tahara, Shigenobu Shibata, Atsuhito Nakao

    Journal of immunology research   2014   474217 - 474217  2014  [Refereed]  [International journal]

     View Summary

    Anaphylaxis is a severe systemic allergic reaction which is rapid in onset and potentially fatal, caused by excessive release of mediators including histamine and cytokines/chemokines from mast cells and basophils upon allergen/IgE stimulation. Increased prevalence of anaphylaxis in industrialized countries requires urgent needs for better understanding of anaphylaxis. However, the pathophysiology of the disease is not fully understood. Here we report that the circadian clock may be an important regulator of anaphylaxis. In mammals, the central clock located in the suprachiasmatic nucleus (SCN) of the hypothalamus synchronizes and entrains peripheral circadian clock present in virtually all cell types via neural and endocrine pathways, thereby driving the daily rhythms in behavior and physiology. We found that mechanical disruption of the SCN resulted in the absence of a time of day-dependent variation in passive systemic anaphylactic (PSA) reaction in mice, associated with loss of daily variations in serum histamine, MCP-1 (CCL2), and IL-6 levels. These results suggest that the central SCN clock controls the time of day-dependent variation in IgE-mediated systemic anaphylactic reaction, which may provide a novel insight into the pathophysiology of anaphylaxis.

    DOI PubMed

  • Warm water bath stimulates phase-shifts of the peripheral circadian clocks in PER2::LUCIFERASE mouse.

    Nobuaki Ohnishi, Yu Tahara, Daisuke Kuriki, Atsushi Haraguchi, Shigenobu Shibata

    PloS one   9 ( 6 ) e100272  2014  [Refereed]  [International journal]

     View Summary

    Circadian clocks in the peripheral tissues of mice are known to be entrained by pulse stimuli such as restricted feeding, novel wheel running, and several other agents. However, there are no reports on high temperature pulse-mediated entrainment on the phase-shift of peripheral clocks in vivo. Here we show that temperature treatment of mice for two days at 41°C, instead of 37°C, for 1-2 h during the inactive period, using a temperature controlled water bath stimulated phase-advance of peripheral clocks in the kidney, liver, and submandibular gland of PER2::LUCIFERASE mice. On the other hand, treatment for 2 days at 35°C ambient room temperature for 2 h did not cause a phase-advance. Maintenance of mice at 41°C in a water bath, sustained the core body temperature at 40-41°C. However, the use of 37°C water bath or the 35°C ambient room temperature elevated the core body temperature to 38.5°C, suggesting that at least a core body temperature of 40-41°C is necessary to cause phase-advance under light-dark cycle conditions. The temperature pulse stimulation at 41°C, instead of 37°C water bath for 2 h led to the elevated expression of Per1 and Hsp70 in the peripheral tissue of mice. In summary, the present study demonstrates that transient high temperature pulse using water bath during daytime causes phase-advance in mouse peripheral clocks in vivo. The present results suggest that hot water bath may affect the phase of peripheral clocks.

    DOI PubMed

  • Effect of quetiapine on Per1, Per2, and Bmal1 clock gene expression in the mouse amygdala and hippocampus.

    Shunpei Moriya, Yu Tahara, Hiroyuki Sasaki, Yutaro Hamaguchi, Daisuke Kuriki, Ryosuke Ishikawa, Jun Ishigooka, Shigenobu Shibata

    Journal of pharmacological sciences   125 ( 3 ) 329 - 32  2014  [Refereed]  [Domestic journal]

     View Summary

    Circadian rhythms are related to various psychiatric disorders. Recently, antipsychotics, including quetiapine (QTP), have been accepted as potential therapeutic agents for the treatment of depression, but its mechanism remains poorly understood. In this study, we examined clock gene fluctuation patterns in QTP-treated mice. QTP significantly increased Per2 mRNA at ZT12 and Per1 and Per2 expression at ZT18 in the amygdala. There were significant differences between the control and QTP groups in the cross-time effects of Per2 mRNA expression in the amygdala. Our findings suggest that QTP possibly acts on the circadian system, which then induces changes in mood symptoms.

    DOI PubMed

  • Acetylcholinesterase (AChE) inhibition aggravates fasting-induced triglyceride accumulation in the mouse liver

    Shin-Ichi Yokota, Kaai Nakamura, Midori Ando, Hiroyasu Kamei, Fumihiko Hakuno, Shin-Ichiro Takahashi, Shigenobu Shibata

    FEBS OPEN BIO   4   905 - 914  2014  [Refereed]

     View Summary

    Although fasting induces hepatic triglyceride (TG) accumulation in both rodents and humans, little is known about the underlying mechanism. Because parasympathetic nervous system activity tends to attenuate the secretion of very-low-density-lipoprotein-triglyceride (VLDL-TG) and increase TG stores in the liver, and serum cholinesterase activity is elevated in fatty liver disease, the inhibition of the parasympathetic neurotransmitter acetylcholinesterase (AChE) may have some influence on hepatic lipid metabolism. To assess the influence of AChE inhibition on lipid metabolism, the effect of physostigmine, an AChE inhibitor, on fasting-induced increase in liver TG was investigated in mice. In comparison with ad libitum-fed mice, 30 h fasting increased liver TG accumulation accompanied by a downregulation of sterol regulatory element-binding protein 1 (SREBP-1) and liver-fatty acid binding-protein (L-FABP). Physostigmine promoted the 30 h fasting-induced increase in liver TG levels in a dose-dependent manner, accompanied by a significant fall in plasma insulin levels, without a fall in plasma TG. Furthermore, physostigmine significantly attenuated the fasting-induced decrease of both mRNA and protein levels of SREBP-1 and L-FABP, and increased IRS-2 protein levels in the liver. The muscarinic receptor antagonist atropine blocked these effects of physostigmine on liver TG, serum insulin, and hepatic protein levels of SREBP-1 and L-FABP. These results demonstrate that AChE inhibition facilitated fasting-induced TG accumulation with up regulation of the hepatic L-FABP and SREBP-1 in mice, at least in part via the activation of muscarinic acetylcholine receptors. Our studies highlight the crucial role of parasympathetic regulation in fasting-induced TG accumulation, and may be an important source of information on the mechanism of hepatic disorders of lipid metabolism. (C) 2014 The Authors. Published by Elsevier B.V. on behalf of the Federation of European Biochemical Societies.

    DOI PubMed

  • Modulation of rat behaviour by using a rat-like robot

    Qing Shi, Hiroyuki Ishii, Shinichi Kinoshita, Atsuo Takanishi, Satoshi Okabayashi, Naritoshi Iida, Hiroshi Kimura, Shigenobu Shibata

    Bioinspiration and Biomimetics   8 ( 4 ) 046002  2013.12  [Refereed]

     View Summary

    In this paper, we study the response of a rat to a rat-like robot capable of generating different types of behaviour (stressful, friendly, neutral). Experiments are conducted in an open-field where a rat-like robot called WR-4 is put together with live rats. The activity level of each rat subject is evaluated by scoring its locomotor activity and frequencies of performing rearing (rising up on its hind limbs) and body grooming (body cuddling and head curling) actions, whereas the degree of preference of that is indicated by the robot-rat distance and the frequency of contacting WR-4. The moving speed and behaviour of WR-4 are controlled in real-time based on the feedback from rat motion. The activity level and degree of preference of rats for each experimental condition are analysed and compared to understand the influence of robot behaviour. The results of this study show that the activity level and degree of preference of the rat decrease when exposed to a stressful robot, and increase when the robot exhibit friendly behaviour, suggesting that a rat-like robot can modulate rat behaviour in a controllable, predictable way. © 2013 IOP Publishing Ltd.

    DOI PubMed

  • Two Distinct Serotonin Receptors Co-mediate Non-photic Signals to the Circadian Clock

    Kazumasa Horikawa, Kazuyuki Fuji, Yuka Fukazawa, Shigenobu Shibata

    JOURNAL OF PHARMACOLOGICAL SCIENCES   123 ( 4 ) 402 - 406  2013.12  [Refereed]

     View Summary

    Several lines of evidence indicate that serotonin type 7 (5-HT7) receptors play a critical role for non-photic resetting of the mammalian circadian clock; however, the contributions of other types of 5-HT receptors to non-photic entrainment are not yet clarified. The present study demonstrates that MKC-242, a selective 5-HT1A receptor agonist, can evoke a non-photic-like phase-response in hamsters in vivo. This phase-shifting response to MKC-242 was antagonized not only by the selective 5-HT1A receptor blocker WAY100635 but also by the selective 5-HT7 receptor blocker DR4004. These suggest that synchronous activation of 5-HT1A and 5-HT7 receptors mediates non-photic signals to the hamster circadian clock.

    DOI PubMed

  • [Chrono-nutrition and chrono-exercise].

    Shibata S, Sasaki H, Ikeda Y

    Nihon rinsho. Japanese journal of clinical medicine   71 ( 12 ) 2194 - 2199  2013.12  [Refereed]  [Domestic journal]

     View Summary

    The circadian rhythm controls many physiological functions, such as feeding, motor activity, endocrine secretion and autonomic nerve. Regular feeding pattern can entrain the peripheral circadian clock, whereas peripheral clock systems can control the absorption distribution, metabolism and excretion of nutrients, suggesting mutual interactions between circadian clocks and nutrition/food. The interactions were so-called by "chrono-nutrition", and bigger meals for breakfast were good for entrainment of peripheral clock and protection of obesity. Similar to chrono-nutrition the timing of exercise ("chrono-exercise") is important for both entrainment signals and energy expenditure. Evening exercise and/or feeding then exercise was good timing exercise for protection of obesity. Taken all, it is suggested that timing of feeding and exercise is now one of key factors for metabolic syndrome.

    PubMed

  • Time-restricted feeding of rapidly digested starches causes stronger entrainment of the liver clock in PER2::LUCIFERASE knock-in mice

    Misa Itokawa, Akiko Hirao, Hiroki Nagahama, Makiko Otsuka, Teiji Ohtsu, Naoki Furutani, Kazuko Hirao, Tamao Hatta, Shigenobu Shibata

    Nutrition Research   33 ( 2 ) 109 - 119  2013.02  [Refereed]

     View Summary

    Restricting feeding to daytime can entrain circadian clocks in peripheral organs of rodents, and nutrients that rapidly increase the blood glucose level are suitable for inducing entrainment. However, dietetic issues, for example, whether or not the diet comprises heated food, have not been fully explored. We therefore hypothesized that rapidly digested starch causes stronger entrainment than slowly digested starch. The entrainment ability of the liver clock in PER2::LUCIFERASE knock-in mice, blood glucose levels, insulin levels, and acute changes in liver clock gene expression were compared between a β-starch (native)-substituted AIN-93M standard diet and an α-starch (gelatinized)-substituted diet. β-Corn and β-rice starch induced larger phase delays of the liver clock, larger blood glucose increases, and higher Per2 gene expression in the liver compared with β-potato starch. Starch granule size, as examined by electron microscopy, was larger for β-potato starch than for β-corn or β-rice starch. After heating, we obtained gelatinized α-potato, α-corn, and α-rice starch, which showed destruction of the crystal structure and a high level of gelatinization. No difference in the increase of blood glucose or insulin levels was observed between β-corn and α-corn starch, or between β-rice and α-rice starch. In contrast, α-potato starch caused higher levels of glucose and insulin compared with β-potato starch. An α-potato starch-substituted diet induced larger phase delays of the liver clock than did β-potato starch. Therefore, rapidly digested starch is appropriate for peripheral clock entrainment. Dietetic issues (heated vs unheated) are important when applying basic mouse data to humans. © 2013 Elsevier Inc.

    DOI PubMed

  • Crosstalk between the circadian clock circuitry and the immune system

    Nicolas Cermakian, Tanja Lange, Diego Golombek, Dipak Sarkar, Atsuhito Nakao, Shigenobu Shibata, Gianluigi Mazzoccoli

    CHRONOBIOLOGY INTERNATIONAL   30 ( 7 ) 870 - 888  2013  [Refereed]

     View Summary

    Various features, components, and functions of the immune system present daily variations. Immunocompetent cell counts and cytokine levels present variations according to the time of day and the sleep-wake cycle. Moreover, different immune cell types, such as macrophages, natural killer cells, and lymphocytes, contain a circadian molecular clockwork. The biological clocks intrinsic to immune cells and lymphoid organs, together with inputs from the central pacemaker of the suprachiasmatic nuclei via humoral and neural pathways, regulate the function of cells of the immune system, including their response to signals and their effector functions. Consequences of this include, for example, the daily variation in the response to an immune challenge (e. g., bacterial endotoxin injection) and the circadian control of allergic reactions. The circadian-immune connection is bidirectional, because in addition to this circadian control of immune functions, immune challenges and immune mediators (e. g., cytokines) were shown to have strong effects on circadian rhythms at the molecular, cellular, and behavioral levels. This tight crosstalk between the circadian and immune systems has wide-ranging implications for disease, as shown by the higher incidence of cancer and the exacerbation of autoimmune symptoms upon circadian disruption.

    DOI PubMed

  • 時間栄養学

    柴田重信, 青木菜摘

    G. I. Research   20 ( 5 )  2012.10

  • 用語解説「時間栄養学」

    柴田重信

    栄養-評価と治療   29 ( 3 )  2012.08

  • Biological clock dysfunction exacerbates contact hypersensitivity in mice.

    Takita E, Yokota S, Tahara Y, Hirao A, Aoki N, Nakamura Y, Nakao A, Shibata S

    Br J Dermatol.    2012.07

    DOI

  • In vivo monitoring of peripheral circadian clocks in the mouse.

    Yu Tahara, Hiroaki Kuroda, Keisuke Saito, Yoshihiro Nakajima, Yuji Kubo, Nobuaki Ohnishi, Yasuhiro Seo, Makiko Otsuka, Yuta Fuse, Yuki Ohura, Takuya Komatsu, Youhei Moriya, Satoshi Okada, Naoki Furutani, Akiko Hirao, Kazumasa Horikawa, Takashi Kudo, Shigenobu Shibata

    Current biology : CB   22 ( 11 ) 1029 - 34  2012.06  [Refereed]  [International journal]

     View Summary

    The mammalian circadian system is comprised of a central clock in the suprachiasmatic nucleus (SCN) and a network of peripheral oscillators located in all of the major organ systems. The SCN is traditionally thought to be positioned at the top of the hierarchy, with SCN lesions resulting in an arrhythmic organism. However, recent work has demonstrated that the SCN and peripheral tissues generate independent circadian oscillations in Per1 clock gene expression in vitro. In the present study, we sought to clarify the role of the SCN in the intact system by recording rhythms in clock gene expression in vivo. A practical imaging protocol was developed that enables us to measure circadian rhythms easily, noninvasively, and longitudinally in individual mice. Circadian oscillations were detected in the kidney, liver, and submandibular gland studied in about half of the SCN-lesioned, behaviorally arrhythmic mice. However, their amplitude was decreased in these organs. Free-running periods of peripheral clocks were identical to those of activity rhythms recorded before the SCN lesion. Thus, we can report for the first time that many of the fundamental properties of circadian oscillations in peripheral clocks in vivo are maintained in the absence of SCN control.

    DOI PubMed

  • 体内時計と食品成分(シリーズ 長寿の秘訣を時計遺伝子から探る)

    柴田重信

    FOODSTYLE21   16 ( 6 )  2012.06

  • Differential roles of breakfast only (one meal per day) and a bigger breakfast with a small dinner (two meals per day) in mice fed a high-fat diet with regard to induced obesity and lipid metabolism.

    Yuta Fuse, Akiko Hirao, Hiroaki Kuroda, Makiko Otsuka, Yu Tahara, Shigenobu Shibata

    Journal of circadian rhythms   10 ( 1 ) 4 - 4  2012.05  [Refereed]  [International journal]

     View Summary

    BACKGROUND: Recent studies on humans and rodents have suggested that the timing of food intake plays an important role in circadian regulation and metabolic health. Consumption of high-fat foods during the inactive period or at the end of the awake period results in weight gain and metabolic syndrome in rodents. However, the distinct effects of breakfast size and the breakfast/dinner size ratio on metabolic health have not yet been fully examined in mice. METHODS: We examined whether the parameters of metabolic syndrome were differentially affected in mice that consumed a large meal at the beginning of the awake period (breakfast; one meal group) and a relatively smaller meal at end of the awake period (dinner; two meals group). The mice of each group were provided equal food volume per day. RESULTS: Mice on one meal exhibited an increase in body weight gain, hyperinsulinemia, hyperleptinemia, and a decrease of gene expression associated with β-oxidation in adipose tissue and liver compared with those on two meals. The circadian expression pattern of the Clock gene in mice on one meal was disturbed compared with those on two meals. CONCLUSIONS: In conclusion, a bigger breakfast with a smaller dinner (two meals per day) but not breakfast only (one meal per day) helps control body weight and fat accumulation in mice on a high-fat meals schedule. The findings of this study suggest that dietary recommendations for weight reduction and/or maintenance should include information on the timing and quantity of dietary intake.

    DOI PubMed

  • 2,2,2-Tribromoethanol phase-shifts the circadian rhythm of the liver clock in Per2::Luciferase knockin mice: lack of dependence on anesthetic activity.

    Yuji Kubo, Yu Tahara, Akiko Hirao, Shigenobu Shibata

    The Journal of pharmacology and experimental therapeutics   340 ( 3 ) 698 - 705  2012.03  [Refereed]  [International journal]

     View Summary

    Comprehensive gene expression profiling in mice in response to the inhalation of sevoflurane has revealed that circadian clock gene expression is affected strongly in the liver, heart, lung, and kidney, in this order, but moderately in the spleen and slightly in the brain. Therefore, we examined whether the administration of general anesthetics at different times of the day induces phase shifts of the liver clock in Per2::Luciferase knockin mice. One to 4 days of intraperitoneal injection of 2,2,2-tribromoethanol (240 mg/kg, anesthetic time 60 min) or 2,2,2-trichloroethanol (240 mg/kg, 60 min), common anesthetics in veterinary surgery, caused phase delays when injected during the daytime and phase advances when injected during the nighttime. Inhalation administration of isoflurane for 30 or 60 min during the daytime did not induce a phase delay. Injection of propofol (300 mg/kg, 17 min) during the daytime induced an insignificant phase delay of the Per2 bioluminescence rhythm. Injection of 2,2,2-tribromoethanol did not induce a phase shift in the suprachiasmatic nucleus, the main oscillator, or in behavioral locomotor rhythms, suggesting that 2,2,2-tribromoethanol induced phase shifts of the liver clock independent of the main suprachiasmatic clock. The expression of clock genes, such as Bmal1 and Clock, in mouse liver was decreased strongly 1 and 4 h after a single injection of 2,2,2-tribromoethanol. These results demonstrate that 2,2,2-tribromoethanol or 2,2,2-trichloroethanol produce phase shifts of the peripheral clock, independent of anesthetic activity. These anesthetics may cause circadian rhythm disorders in peripheral organs when administered as general anesthetics several times during the day.

    DOI PubMed

  • Involvement of Stress Kinase Mitogen-activated Protein Kinase Kinase 7 in Regulation of Mammalian Circadian Clock

    Yoshimi Uchida, Tomomi Osaki, Tokiwa Yamasaki, Tadanori Shimomura, Shoji Hata, Kazumasa Horikawa, Shigenobu Shibata, Takeshi Todo, Jun Hirayama, Hiroshi Nishina

    JOURNAL OF BIOLOGICAL CHEMISTRY   287 ( 11 ) 8318 - 8326  2012.03  [Refereed]

     View Summary

    The stress kinase mitogen-activated protein kinase kinase 7 (MKK7) is a specific activator of c-Jun N-terminal kinase (JNK), which controls various physiological processes, such as cell proliferation, apoptosis, differentiation, and migration. Here we show that genetic inactivation of MKK7 resulted in an extended period of oscillation in circadian gene expression in mouse embryonic fibroblasts. Exogenous expression in cultured mammalian cells of an MKK7-JNK fusion protein that functions as a constitutively active form of JNK induced phosphorylation of PER2, an essential circadian component. Furthermore, JNK interacted with PER2 at both the exogenous and endogenous levels, and MKK7-mediated JNK activation increased the half-life of PER2 protein by inhibiting its ubiquitination. Notably, the PER2 protein stabilization induced by MKK7-JNK fusion protein reduced the degradation of PER2 induced by casein kinase 1 epsilon. Taken together, our results support a novel function for the stress kinase MKK7 as a regulator of the circadian clock in mammalian cells at steady state.

    DOI PubMed

  • 時計遺伝子と食事リズム-時間栄養学-

    柴田重信

    日本臨床栄養学会誌    2012

  • 概日リズムと肥満・生活習慣病「体内時計と栄養・食事の相互作用」

    日本肥満学会誌    2012

  • 生体リズムと創薬

    柴田重信

    日本臨床薬理学会誌    2012

  • 時間栄養学

    柴田重信

    化学と生物    2012

  • 暮らしの最前線90「時間栄養学の現状とこれから」

    柴田重信

    日本家政学会誌   63 ( 6 ) 337 - 341  2012

    DOI

  • 体内時計を用いた理想的な食生活作りのために

    平尾彰子, 柴田重信

    顕微鏡   47 ( 2 ) 83 - 86  2012

  • The Protective and Recovery Effects of Fish Oil Supplementation on Cedar Pollen-Induced Allergic Reactions in Mice.

    Hirao A, Furutani N, Nagahama H, Itokawa M, Shibata S

    Food Nutrition and Sciences   3   40 - 47  2012.01

  • Meal frequency patterns determine the phase of mouse peripheral circadian clocks.

    Hiroaki Kuroda, Yu Tahara, Keisuke Saito, Nobuaki Ohnishi, Yuji Kubo, Yasuhiro Seo, Makiko Otsuka, Yuta Fuse, Yuki Ohura, Akiko Hirao, Shigenobu Shibata

    Scientific reports   2   711 - 711  2012  [Refereed]  [International journal]

     View Summary

    Peripheral circadian clocks in mammals are strongly entrained by light-dark and eating cycles. Their physiological functions are maintained by the synchronization of the phase of organs via clock gene expression patterns. However, little is known about the adaptation of peripheral clocks to the timing of multiple daily meals. Here, we investigated the effect of irregular eating patterns, in terms of timing and volume, on their peripheral clocks in vivo. We found that the phase of the peripheral clocks was altered by the amount of food and the interval between feeding time points but was unaffected by the frequency of feeding, as long as the interval remained fixed. Moreover, our results suggest that a late dinner should be separated into 2 half-dinners in order to alleviate the effect of irregular phases of peripheral clocks.

    DOI PubMed

  • Stress exposure using small mobile robot both in immature and mature period induces mental disorder in rat

    Hiroyuki Ishii, Qing Shi, Syunsuke Miyagishima, Shogo Fumino, Shinichiro Konno, Satoshi Okabayashi, Naritoshi Iida, Hiroshi Kimura, Yu Tahara, Shigenobu Shibata, Atsuo Takanishi

    Proceedings of the IEEE RAS and EMBS International Conference on Biomedical Robotics and Biomechatronics     587 - 592  2012  [Refereed]

     View Summary

    The number of patients with mental disorders is increasing in advanced countries, hence more effective psychotropic drugs are recently desired. In process of development of psychotropic drugs, animal experiments have been playing a very important role. Mental disorder model animals which exhibit behavior disorder like patients with mental disorders are used in these experiments. These animals are normally developed by genetic manipulation, surgical operation in their brain or drug administration. A candidate for a new drug is administrated in these animals to evaluate its effect. However, we have some doubts about conventional mental disorder model animals because they are induced these disorders by using methods which are quite different from causes of mental disorder of human beings. Therefore, the purpose of this study is to develop an novel methodology to create mental disorder model animals. We then developed a small mobile robot and a control system for it. Using them, we have performed some experiments to create a mental disorder model rat. We had then succeeded in developing a mental disorder model rat by exposing stress using the robot during immature period. This rat exhibits low activity in some behavior tests during mature period. For better understanding of how stress exposure induces mental disorder in a rat, we conducted another experiment based on stress vulnerability hypothesis. In this experiment, stress was exposed during both immature and mature period while that had been exposed only during immature period. We prepared several conditions of stress exposure by changing robot behavior pattern to find the one to induce much stress in a rat. From a result of experiment, we found that a rat which received gentle chase by the robot during immature period was induced much stress when it received robot attack during mature period. Thus, we consider that this rat is more appropriate to the mental disorder model than that was developed in our past experiment. © 2012 IEEE.

    DOI

  • Stress Exposure using Small Mobile Robot both in Immature and Mature Period Induces Mental Disorder in Rat

    Hiroyuki Ishii, Qing Shi, Syunsuke Miyagishima, Shogo Fumino, Shinichiro Konno, Satoshi Okabayashi, Naritoshi Iida, Hiroshi Kimura, Yu Tahara, Shigenobu Shibata, Atsuo Takanishi

    2012 4TH IEEE RAS & EMBS INTERNATIONAL CONFERENCE ON BIOMEDICAL ROBOTICS AND BIOMECHATRONICS (BIOROB)     587 - 592  2012  [Refereed]

     View Summary

    The number of patients with mental disorders is increasing in advanced countries, hence more effective psychotropic drugs are recently desired. In process of development of psychotropic drugs, animal experiments have been playing a very important role. Mental disorder model animals which exhibit behavior disorder like patients with mental disorders are used in these experiments. These animals are normally developed by genetic manipulation, surgical operation in their brain or drug administration. A candidate for a new drug is administrated in these animals to evaluate its effect. However, we have some doubts about conventional mental disorder model animals because they are induced these disorders by using methods which are quite different from causes of mental disorder of human beings. Therefore, the purpose of this study is to develop an novel methodology to create mental disorder model animals. We then developed a small mobile robot and a control system for it. Using them, we have performed some experiments to create a mental disorder model rat. We had then succeeded in developing a mental disorder model rat by exposing stress using the robot during immature period. This rat exhibits low activity in some behavior tests during mature period. For better understanding of how stress exposure induces mental disorder in a rat, we conducted another experiment based on stress vulnerability hypothesis. In this experiment, stress was exposed during both immature and mature period while that had been exposed only during immature period. We prepared several conditions of stress exposure by changing robot behavior pattern to find the one to induce much stress in a rat. From a result of experiment, we found that a rat which received gentle chase by the robot during immature period was induced much stress when it received robot attack during mature period. Thus, we consider that this rat is more appropriate to the mental disorder model than that was developed in our past experiment.

  • Synthesis of a New [6]-Gingerol Analogue and Its Protective Effect with Respect to the Development of Metabolic Syndrome in Mice Fed a High-Fat Diet

    Mayumi Okamoto, Hiroyuki Irii, Yu Tahara, Hiroyuki Ishii, Akiko Hirao, Haruhide Udagawa, Masaki Hiramoto, Kazuki Yasuda, Atsuo Takanishi, Shigenobu Shibata, Isao Shimizu

    JOURNAL OF MEDICINAL CHEMISTRY   54 ( 18 ) 6295 - 6304  2011.09  [Refereed]

     View Summary

    To determine the effects of a [6]-gingerol analogue (6G), a major chemical component of the ginger rhizome, and its stable analogue after digestion in simulated gastric fluid, aza-[6]-gingerol (A6G), on diet-induced body fat accumulation, we synthesized 6G and A6G. Mice were fed either a control regular rodent chow, a high-fat diet (HFD), or a HFD supplemented with 6G and A6G. Magnetic resonance imaging adiposity parameters of the 6G- and A6G-treated mice were compared with those of control mice. Supplementation with 6G and A6G significantly reduced body weight gain, fat accumulation, and circulating levels of insulin and leptin. The mRNA levels of sterol regulatory element-binding protein 1c (SREBP-1c) and acetyl-CoA carboxylase 1 in the liver were significantly lower in mice fed A6G than in HFD control mice. Our findings indicate that A6G, rather than 6G, enhances energy metabolism and reduces the extent of lipogenesis by downregulating SREBP-1c and its related molecules, which leads to the suppression of body fat accumulation.

    DOI PubMed

  • Dietary intake is associated with human chronotype as assessed by both morningness-eveningness score and preferred midpoint of sleep in young Japanese women

    Natsuko Sato-Mito, Shigenobu Shibata, Satoshi Sasaki, Kazuto Sato

    INTERNATIONAL JOURNAL OF FOOD SCIENCES AND NUTRITION   62 ( 5 ) 525 - 532  2011.08  [Refereed]

     View Summary

    We analyzed the association between dietary intake and chronotype as assessed by both Morningness-Eveningness Questionnaire (MEQ) score and preferred midpoint of sleep in 112 young Japanese women. Dietary intake was assessed by a brief, self-administered diet history questionnaire. A lower MEQ score (evening-type tendency) showed a significant association with a lower energy-adjusted intake of protein, calcium, magnesium, zinc, vitamins (D, riboflavin, and B(6)), and vegetables, and with a higher intake of noodles. Furthermore, a later midpoint of sleep showed a significant association with a lower energy-adjusted intake of protein, cholesterol, potassium, calcium, magnesium, zinc, vitamins (D, riboflavin, B(6), and B(12)), soy, fish and shellfish, and eggs, and with a higher intake of noodles, bread, and confections. These data suggest that evening chronotype is associated with inadequate dietary habits such as low vitamin and mineral intakes.

    DOI PubMed

  • Refeeding after fasting elicits insulin-dependent regulation of Per2 and Rev-erbα with shifts in the liver clock.

    Yu Tahara, Makiko Otsuka, Yuta Fuse, Akiko Hirao, Shigenobu Shibata

    Journal of biological rhythms   26 ( 3 ) 230 - 40  2011.06  [Refereed]  [International journal]

     View Summary

    The mammalian circadian clock is known to be entrained by both a daily light-dark cycle and daily feeding cycle. However, the mechanisms of feeding-induced entrainment are not as fully understood as those of light entrainment. To elucidate the first step of entrainment of the liver clock, we identified the circadian clock gene(s) that show both phase advance and acute change of gene expression during the early term of the daytime refeeding schedule in mice. The expressions of liver Per2 and Rev-erbα genes were phase-advanced within 1 day of refeeding. Additionally, the upregulation of Per2 mRNA and down-regulation of Rev-erbα mRNA were induced within 2 hours, not only by food intake but also by insulin injection in intact mice. These expression changes by food intake were not revealed in streptozotocin-treated insulin-deficient mice, but insulin injection was able to recover the impairment of Per2 and Rev-erbα gene expression. Furthermore, we demonstrated using an ex vivo luciferase monitoring system that insulin injection during the daytime causes a phase advance of liver Per2 expression rhythm in Per2::luciferase knock-in mice. In embryonic fibroblasts from Per2::luciferase knock-in mice, insulin infusion caused an acute increase of Per2 gene expression and a similar phase advance of Per2 expression rhythm. Our results indicate that an acute change of Per2 and Rev-erbα gene expression mediated by refeeding-induced insulin secretion is a critical step mediating the early phase of feeding-induced entrainment of the liver clock.

    DOI PubMed

  • Circadian clock gene Period2 regulates a time-of-day-dependent variation in cutaneous anaphylactic reaction.

    Yuki Nakamura, Daisuke Harama, Naomi Shimokawa, Mutsuko Hara, Ryuyo Suzuki, Yu Tahara, Kayoko Ishimaru, Ryohei Katoh, Ko Okumura, Hideoki Ogawa, Shigenobu Shibata, Atsuhito Nakao

    The Journal of allergy and clinical immunology   127 ( 4 ) 1038 - 45  2011.04  [Refereed]  [International journal]

     View Summary

    BACKGROUND: IgE-mediated immediate-type skin reaction shows a diurnal rhythm, although the precise mechanisms remain uncertain. Period2 (Per2) is a key circadian gene that is essential for endogenous clockworks in mammals. OBJECTIVE: This study investigated whether Per2 regulates a time-of-day-dependent variation in IgE-mediated immediate-type skin reaction. METHODS: The kinetics of a passive cutaneous anaphylactic reaction were compared between wild-type mice and mice with a loss-of-function mutation of Per2 (mPer2(m/m) mice). The effects of adrenalectomy, aging, and dexamethasone on the kinetics of a passive cutaneous anaphylactic reaction were also examined. In addition, the extent of IgE-mediated degranulation in bone marrow-derived mast cells (BMMCs) was compared between wild-type and mPer2(m/m) mice. RESULTS: A time-of-day-dependent variation in a passive cutaneous anaphylactic reaction observed in wild-type mice was absent in mPer2(m/m) mice and in adrenalectomized and aged mice associated with the loss of rhythmic secretion of corticosterone. In addition, mPer2(m/m) mice showed decreased sensitivity to the inhibitory effects of dexamethasone on the passive cutaneous anaphylactic reactions. IgE-mediated degranulation in BMMCs was comparable between wild-type and mPer2(m/m) mice, but Per2 mutation decreased sensitivity to the inhibitory effects of dexamethasone on IgE-mediated degranulation in BMMCs. CONCLUSION: A circadian oscillator, Per2, regulates a time-of-day-dependent variation in a passive cutaneous anaphylactic reaction in mice. Per2 may do so by controlling the rhythmic secretion of glucocorticoid from adrenal glands and/or by gating the glucocorticoid responses of mast cells to certain times of the day (possibly when Per2 levels are high in mast cells).

    DOI PubMed

  • Basic and applied science of chrononutrition.

    Shibata S, Hirao A

    Nihon yakurigaku zasshi. Folia pharmacologica Japonica   137 ( 3 ) 110 - 114  2011.03  [Refereed]

    DOI PubMed

  • Attenuated Food Anticipatory Activity and Abnormal Circadian Locomotor Rhythms in Rgs16 Knockdown Mice

    Naoto Hayasaka, Kazuyuki Aoki, Saori Kinoshita, Shoutaroh Yamaguchi, John K. Wakefield, Sachiyo Tsuji-Kawahara, Kazumasa Horikawa, Hiroshi Ikegami, Shigeharu Wakana, Takamichi Murakami, Ram Ramabhadran, Masaaki Miyazawa, Shigenobu Shibata

    PLOS ONE   6 ( 3 ) e17655  2011.03  [Refereed]

     View Summary

    Regulators of G protein signaling (RGS) are a multi-functional protein family, which functions in part as GTPase-activating proteins (GAPs) of G protein alpha-subunits to terminate G protein signaling. Previous studies have demonstrated that the Rgs16 transcripts exhibit robust circadian rhythms both in the suprachiasmatic nucleus (SCN), the master circadian light-entrainable oscillator (LEO) of the hypothalamus, and in the liver. To investigate the role of RGS16 in the circadian clock in vivo, we generated two independent transgenic mouse lines using lentiviral vectors expressing short hairpin RNA (shRNA) targeting the Rgs16 mRNA. The knockdown mice demonstrated significantly shorter free-running period of locomotor activity rhythms and reduced total activity as compared to the wild-type siblings. In addition, when feeding was restricted during the daytime, food-entrainable oscillator (FEO)-driven elevated food-anticipatory activity (FAA) observed prior to the scheduled feeding time was significantly attenuated in the knockdown mice. Whereas the restricted feeding phase-advanced the rhythmic expression of the Per2 clock gene in liver and thalamus in the wild-type animals, the above phase shift was not observed in the knockdown mice. This is the first in vivo demonstration that a common regulator of G protein signaling is involved in the two separate, but interactive circadian timing systems, LEO and FEO. The present study also suggests that liver and/or thalamus regulate the food-entrained circadian behavior through G protein-mediated signal transduction pathway(s).

    DOI PubMed

  • The midpoint of sleep is associated with dietary intake and dietary behavior among young Japanese women

    Natsuko Sato-Mito, Satoshi Sasaki, Kentaro Murakami, Hitomi Okubo, Yoshiko Takahashi, Shigenobu Shibata, Kazuhiko Yamada, Kazuto Sato

    SLEEP MEDICINE   12 ( 3 ) 289 - 294  2011.03  [Refereed]

     View Summary

    Objectives: How human chronotype is correlated to nutrient and food-group intakes and dietary behavior remains to be elucidated. We cross-sectionally examined the association between the midpoint of sleep and these dietary variables in young Japanese women. A calculated halfway point between bedtime and rise time was used as midpoint of sleep.
    Methods: The subjects were 3304 female Japanese dietetics students aged 18-20 years from 53 institutions in Japan. Dietary intake during the previous month was assessed by a validated, self-administered diet history questionnaire. The midpoint of sleep was calculated using self-reported bedtimes and rise times.
    Results: Late midpoint of sleep was significantly negatively associated with the percentage of energy from protein and carbohydrates, and the energy-adjusted intake of cholesterol, potassium, calcium, magnesium, iron, zinc, vitamin A, vitamin D, thiamin, riboflavin, vitamin B(6), folate, rice, vegetables, pulses, eggs, and milk and milk products. It was also significantly positively associated with the percentage of energy from alcohol and fat, and the energy-adjusted intake of noodles, confections, fat and oil, and meat. Furthermore, subjects with a later midpoint of sleep tended to begin meals later, eat for a longer time, skip meals more frequently, and watch TV at meals, not only at breakfast but also at lunch and dinner.
    Conclusions: The midpoint of sleep is significantly associated with dietary intake of certain nutrients and foods and other dietary behaviors in young Japanese women. This finding may contribute to consider the relationships between chronotype and dietary intakes and behaviors. (C) 2011 Elsevier B.V. All rights reserved.

    DOI PubMed

  • Effects of ATP on circadian rhythm of Per2 expression in astrocytes

    Kobayashi Kei, Shibata Shigenobu, Koizumi Schuichi

    JOURNAL OF PHARMACOLOGICAL SCIENCES   115   243P  2011  [Refereed]

  • Combination of starvation interval and food volume determines the phase of liver circadian rhythm in Per2::Luc knock-in mice under two meals per day feeding.

    Akiko Hirao, Hiroki Nagahama, Takuma Tsuboi, Mizuho Hirao, Yu Tahara, Shigenobu Shibata

    American journal of physiology. Gastrointestinal and liver physiology   299 ( 5 ) G1045-53 - 1053  2010.11  [Refereed]  [International journal]

     View Summary

    Although the circadian liver clock is entrained by the feeding cycle, factors such as food volume and starvation interval are poorly understood. Per2::Luc knock-in mice were given two meals per day with different food volume sizes and/or with different intervals of starvation between two mealtimes with the total food volume per day fixed at 3.6 g (80 food pellets, ∼75% of free feeding) per mouse. The bioluminescence rhythm in the liver produced a unimodal peak but not bimodal peak under the regimen of two meals per day over 14-15 days. Peak Per2 expression occurred concurrently with the mealtime of the larger food volume, when the first and second meal were given as different food volume ratios under a 12 h feeding interval. When an equal volume of food was given under different starvation interval (8 h:16 h), the peak of the Per2 rhythm was close to peak by mealtime after long starvation (16 h). When food volumes for each mealtime were changed under 8 h:16 h, the peak rhythm was influenced by combined factors of food volume and starvation interval. Food intake after the 16-h starvation caused a significant increase in liver Per2, Dec1, and Bmal1 gene expression compared with food intake after the 8-h starvation with 8 h:16 h feeding intervals. In conclusion, the present results clearly demonstrate that food-induced entrainment of the liver clock is dependent on both food volume and the starvation interval between two meals. Therefore, normal feeding habits may help to maintain normal clock function in the liver organ.

    DOI PubMed

  • Time of Day and Nutrients in Feeding Govern Daily Expression Rhythms of the Gene for Sterol Regulatory Element-binding Protein (SREBP)-1 in the Mouse Liver

    Eriko Matsumoto, Akinori Ishihara, Saki Tamai, Ayako Nemoto, Katsuro Iwase, Takaki Hiwasa, Shigenobu Shibata, Masaki Takiguchi

    JOURNAL OF BIOLOGICAL CHEMISTRY   285 ( 43 ) 33028 - 33036  2010.10  [Refereed]

     View Summary

    Sterol regulatory element-binding protein-1 (SREBP-1) plays a central role in transcriptional regulation of genes for hepatic lipid synthesis that utilizes diet-derived nutrients such as carbohydrates and amino acids, and expression of SREBP-1 exhibits daily rhythms with a peak in the nocturnal feeding period under standard housing conditions of mice. Here, we report that the Srebp-1 expression rhythm shows time cue-independent and Clock mutation-sensitive circadian nature, and is synchronized with varied photoperiods apparently through entrainment of locomotor activity and food intake. Fasting caused diminution of Srebp-1 expression, while diabetic db/db and ob/ob mice showed constantly high expression with loss of rhythmicity. Time-restricted feedings during mid-light and mid-dark periods exhibited differential effects, the latter causing more severe damping of the oscillation. Therefore, "when to eat in a day (the light/dark cycle)," rather than "whenever to eat in a day," is a critical determinant to shape the daily rhythm of Srebp-1 expression. We further found that a high-carbohydrate diet and a high-protein diet, as well as a high-fat diet, cause phase shifts of the oscillation peak into the light period, underlining the importance of "what to eat." Daily rhythms of SREBP-1 protein levels and Akt phosphorylation levels also exhibited nutrient-responsive changes. Taken together, these findings provide a model for mechanisms by which time of day and nutrients in feeding shape daily rhythms of the Srebp-1 expression and possibly a number of other physiological functions with interindividual and interdaily differences in human beings and wild animals subjected to day-by-day changes in dietary timing and nutrients.

    DOI PubMed

  • The adjustment and manipulation of biological rhythms by light, nutrition, and abused drugs.

    Shigenobu Shibata, Yu Tahara, Akiko Hirao

    Advanced drug delivery reviews   62 ( 9-10 ) 918 - 27  2010.07  [Refereed]  [International journal]

     View Summary

    Daily restricted feeding entrains the circadian rhythm of mouse clock gene expression in the central nervous system, excluding the suprachiasmatic nucleus (SCN), as well as in the peripheral tissues such as the liver, lung, and heart. In addition to entrainment of the clock genes, daily restricted feeding induces a locomotor activity increase 2-3h before the restricted feeding time initiates. The increase in activity is called the food-anticipatory activity (FAA). In addition to FAA, daily restricted feeding can also entrain peripheral circadian clocks in other organs such as liver, lung, and heart. This type of oscillator is called the food-entrainable peripheral oscillator (FEPO). At present, the mechanisms for restricted feeding-induced entrainment of locomotor activity (FAA) and/or peripheral clock (FEPO) are still unknown. In this review, we describe the role of the central nervous system and peripheral tissues in FAA performance and also in the entrainment of clock gene expression. In addition, the mechanism for entrainment of circadian oscillators by the abuse of drugs, such as methamphetamine, is discussed.

    DOI PubMed

  • Cortical neurons from intrauterine growth retardation rats exhibit lower response to neurotrophin BDNF

    Midori Ninomiya, Tadahiro Numakawa, Naoki Adachi, Miyako Furuta, Shuichi Chiba, Misty Richards, Shigenobu Shibata, Hiroshi Kunugi

    NEUROSCIENCE LETTERS   476 ( 2 ) 104 - 109  2010.05  [Refereed]

     View Summary

    Intrauterine growth retardation (IUGR) is putatively involved in the pathophysiology of schizophrenia. The animal model of IUGR induced by synthetic thromboxane A2 (TXA2) is useful to clarify the effect of IUGR on pups' brains, however, analysis at the cellular level is still needed. Brain-derived neurotrophic factor (BDNF), which plays a role in neuronal survival and synaptic plasticity in the central nervous system (CNS), may also be associated with schizophrenia. However, the possible relationship between IUGR and BDNF function remains unclear. Here, we examined how IUGR by TXA2 impacts BDNF function by using dissociated cortical neurons. We found that, although BDNF levels in cultured neurons from the cerebral cortex of low birth weight pups with IUGR were unchanged, TrkB (BDNF receptor) was decreased compared with control-rats. BDNF-stimulated MAPK/ERK1/2 and PI3K/Akt pathways, which are downstream intracellular signaling pathways of TrkB, were repressed in IUGR-rat cultures. Expression of glutamate receptors such as GluA1 and GluN2A was also suppressed in IUGR-rat cultures. Furthermore, in IUGR-rat cultures, anti-apoptotic protein Bcl2 was decreased and BDNF failed to prevent neurons from cell death caused by serum-deprivation. Taken together, IUGR resulted in reductions in cell viability and in synaptic function following TrkB down-regulation, which may play a role in schizophrenia-like behaviors. (C) 2010 Elsevier Ireland Ltd. All rights reserved.

    DOI PubMed

  • 体内時計の活用法

    柴田重信

    明治学院大学教養教育センター付属研究所年報(Synthesis 2009)     29 - 37  2010.03

  • Self-sustained circadian rhythm in cultured human mononuclear cells isolated from peripheral blood

    Takashi Ebisawa, Kahori Numazawa, Hiroko Shimada, Hiroyuki Izutsu, Tsukasa Sasaki, Nobumasa Kato, Katsushi Tokunaga, Akio Mori, Ken-ichi Honma, Sato Honma, Shigenobu Shibata

    NEUROSCIENCE RESEARCH   66 ( 2 ) 223 - 227  2010.02  [Refereed]

     View Summary

    Disturbed circadian rhythmicity is associated with human diseases such as sleep and mood disorders. However, study of human endogenous circadian rhythm is laborious and time-consuming, which hampers the elucidation of diseases. It has been reported that peripheral tissues exhibit circadian rhythmicity as the suprachiasmatic nucleus-the center of the biological clock. We tried to study human circadian rhythm using cultured peripheral blood mononuclear cells (PBMCs) obtained from a single collection of venous blood. Activated human PBMCs showed self-sustained circadian rhythm of clock gene expression, which indicates that they are useful for investigating human endogenous circadian rhythm. (C) 2009 Elsevier Ireland Ltd and the Japan Neuroscience Society. All rights reserved.

    DOI PubMed

  • Effects of medial hypothalamic lesions on feeding-induced entrainment of locomotor activity and liver Per2 expression in Per2::luc mice.

    Yu Tahara, Akiko Hirao, Takahiro Moriya, Takashi Kudo, Shigenobu Shibata

    Journal of biological rhythms   25 ( 1 ) 9 - 18  2010.02  [Refereed]  [International journal]

     View Summary

    Restricted feeding induces anticipatory activity rhythm and also entrains the peripheral circadian clocks, although the underlying brain mechanisms have not been fully elucidated. The dorsomedial hypothalamus (DMH) has been implicated in the regulation of restricted feeding-induced anticipatory activity rhythms (FAA), but the role of the DMH in restricted feeding- induced entrainment of peripheral circadian clocks is still unknown. In the present study, the role of the DMH in entrainment of the peripheral circadian clock was examined using Per2::luciferase knock-in mice. The results indicate that lesions that destroy the large mediobasal hypothalamic (MBH) lesions destroying the DMH, ventrolateral hypothalamus (VMH), and arcuate nucleus (ARC) significantly reduce daily locomotor activity rhythms and FAA formation. In addition, these lesions phase advanced the peak of liver Per2 expression by 2 h when compared to sham-operated mice. Following the administration of MBH lesions, the animals run less and start later in the restricted feeding- induced FAA rhythm but do not have any alterations in the restricted feeding- induced phase shift of the liver Per2 rhythm. These results demonstrate that the hypothalamus, including the MBH, is an important brain area for maintaining the locomotor rhythm and FAA formation. However, it is not necessary for restricted feeding-induced entrainment of the liver clock.

    DOI PubMed

  • Prednisolone causes anxiety- and depression-like behaviors and altered expression of apoptotic genes in mice hippocampus

    Yu Kajiyama, Yoshimi Iijima, Shuichi Chiba, Miyako Furuta, Midori Ninomiya, Aiko Izumi, Shigenobu Shibata, Hiroshi Kunugi

    PROGRESS IN NEURO-PSYCHOPHARMACOLOGY & BIOLOGICAL PSYCHIATRY   34 ( 1 ) 159 - 165  2010.02  [Refereed]

     View Summary

    Glucocorticoids are known to cause psychiatric disorders including depression. Prednisolone (PSL) is one of the most widely used synthetic glucocorticoids to treat various medical diseases; however, little is known about PSL-induced behavioral changes and its molecular basis in the brain. Growing evidence has implicated that hippocampal remodeling or damage play a role in the pathogenic effect of glucocorticoids. In this study, mice were administered PSL (50 or 100 mg/kg) or vehicle for 6 or 7 days and subjected to a series of behavioral tests, i.e., open field, elevated plus maze, prepulse inhibition, forced swim, and tail suspension tests. Hippocampal tissues were subject to microarray analysis using the GeneChip Mouse Genome 430 2.0 Array (Affymetrix) containing 45,101 probes of transcripts. Increased anxiety- and depression-like behaviors assessed with open field, elevated plus maze, and tail suspension tests were observed. Microarray analysis detected 108 transcripts with a fold change of &gt;2.0 or &lt;0.5 in which many cell-death-related genes were found. The microarray data was validated by quantitative reverse transcriptase-polymerase chain reaction analysis. Our results demonstrated that PSL causes anxiety- and depression-like behaviors, and suggest that altered gene expressions related to hippocampal remodeling or damage are involved in the effect of PSL on such behaviors. (C) 2009 Elsevier Inc. All rights reserved.

    DOI PubMed

  • The role of GABAergic neuron on NMDA- and SP-induced phase delays in the suprachiasmatic nucleus neuronal activity rhythm in vitro

    Toshiyuki Hamada, Shigenobu Shibata

    NEUROSCIENCE LETTERS   468 ( 3 ) 344 - 347  2010.01  [Refereed]

     View Summary

    Gamma-aminobutyric acid (GABA), and its biosynthetic enzyme, glutamic decarboxylase, are widely distributed in the suprachiasmatic nucleus (SCN). In the present study, we examined the role of the GABA(A) receptor on in vitro SCN responses to photic-like signals. We found that 100 mu M GABA(A) receptor antagonist bicuculline partially blocked field potentials evoked by optic nerve stimulation. NMDA- and SP-induced phase shifts of SCN neuronal activity rhythms, were blocked with 10 mu M bicuculline. Application of 100 mu M bicuculline alone induced phase advance of SCN neuronal activity rhythm. These results show that NMDA- and SP-induced phase shifts are blocked by bicuculline and suggest GABA has an important role as neurotransmitter in the neuronal network regulating phase shifts of the circadian clock. (C) 2009 Elsevier Ireland Ltd. All rights reserved.

    DOI PubMed

  • Restricted feeding-induced entrainment of activity rhythm and peripheral clock rhythm

    Shigenobu Shibata, Akiko Hirao, Yu Tahara

    SLEEP AND BIOLOGICAL RHYTHMS   8 ( 1 ) 18 - 27  2010.01

     View Summary

    Daily restricted feeding entrains the circadian rhythm of mouse clock gene expression in the central nervous system excluding the suprachiasmatic nucleus, as well as in the peripheral tissues such as the liver, lungs, and heart. In addition to entrainment of the clock gene, daily restricted feeding induces a locomotor activity increase 2-3 h before the restricted feeding time. The increase of activity is called the food anticipatory activity (FAA). At present, the mechanisms for restricted feeding-induced entrainment are still unknown. In this review, we describe the role of the central nervous system and peripheral tissues in FAA performance and also in entrainment of clock gene expression.

    DOI

  • TIME-DEPENDENT INHIBITORY EFFECT OF LIPOPOLYSACCHARIDE INJECTION ON PER1 AND PER2 GENE EXPRESSION IN THE MOUSE HEART AND LIVER

    Yusuke Yamamura, Ichiro Yano, Takashi Kudo, Shigenobu Shibata

    CHRONOBIOLOGY INTERNATIONAL   27 ( 2 ) 213 - 232  2010  [Refereed]

     View Summary

    Lipopolysaccharide (LPS) is a pathogen-associated large molecule responsible for sepsis-related endotoxic shock, and the heart is one of the most common organs adversely affected. LPS is reported to increase serum TNF alpha levels and reduce Per1 and Per2 gene expression. Therefore, in this experiment, we determined the time-dependent effects of LPS on heart rate (HR) and circadian gene expression in the mouse heart and liver. HR of the LPS group was significantly elevated 2 and 8 h after injection compared to the control group. A significant percent increase in HR was observed at ZT6, 12, and 18. LPS increased Tnf alpha mRNA expression in the heart and liver at ZT6, 18, and 24. A time-dependent effect of LPS on reduction of Per1 and Per2 gene expression was also observed in the heart and liver. In order to examine the effect of LPS on cell damage, we examined apoptosis-related gene expression after LPS injection. Bax mRNA expression level of the LPS group was higher than that of the control group 8 and 26 h after injection. On the other hand, Bcl2 mRNA expression level of the LPS group was lower than that of the control group 2 and 26 h after injection. Dexamethasone strongly attenuated the LPS-induced increase of serum TNF alpha without significant change in Per1 and Per2 gene expression in the heart. In conclusion, the present results demonstrated that LPS exerts a time-dependent inhibitory effect on Per1 and Per2 gene expression in the heart and liver. The chronopharmacological lethal effect of LPS may be related to the time-dependent increase of serum TNF alpha level and simultaneously high level of Per2 gene expression in the heart and liver between ZT12-18. Taken together, chronopharmacological effect of LPS may be related to not only sickness behavior syndrome and mortality, but also circadian rhythm systems. (Author correspondence: shibatas@waseda.jp).

    DOI PubMed

  • Cultured neurons from intrauterine growth retardation rats display lower response to brain-derived neurotrophic factors

    Midori Ninomiya, Tadahiro Numakawa, Naoki Adachi, Miyako Furuta, Shuichi Chiba, Misty Richards, Shigenobu Shibata, Hiroshi Kunugi

    NEUROSCIENCE RESEARCH   68   E258 - E258  2010  [Refereed]

    DOI

  • Possible changes in neurotrophin signalings and levels of ER alpha are involved in anxiety- and depressive- like behaviors in postpartum rats after weaning

    Furuta Miyako, Numakawa Tadahiro, Ninomiya Midori, Chiba Shuichi, Kajiyama Yu, Shibata Shigenobu, Funabashi Toshiya, Akema Tatsuo, Kunugi Hiroshi

    NEUROSCIENCE RESEARCH   68   E285  2010  [Refereed]

    DOI

  • A balanced diet is necessary for proper entrainment signals of the mouse liver clock.

    Akiko Hirao, Yu Tahara, Ichiro Kimura, Shigenobu Shibata

    PloS one   4 ( 9 ) e6909  2009.09  [Refereed]  [International journal]

     View Summary

    BACKGROUND: The peripheral circadian clock in mice is entrained not only by light-dark cycles but also by daily restricted feeding schedules. Behavioral and cell culture experiments suggest an increase in glucose level as a factor in such feeding-induced entrainment. For application of feeding-induced entrainment in humans, nutrient content and dietary variations should be considered. PRINCIPAL FINDING: To elucidate the food composition necessary for dietary entrainment, we examined whether complete or partial substitution of dietary nutrients affected phase shifts in liver clocks of mice. Compared with fasting mice or ad libitum fed mice, the liver bioluminescence rhythm advanced by 3-4 h on the middle day in Per2::luciferase knock-in mice that were administered a standard mouse diet, i.e. AIN-93M formula [0.6-0.85 g/10 g mouse BW] (composition: 14% casein, 47% cornstarch, 15% gelatinized cornstarch, 10% sugar, 4% soybean oil, and 10% other [fiber, vitamins, minerals, etc.]), for 2 days. When each nutrient was tested alone (100% nutrient), an insignificant weak phase advance was found to be induced by cornstarch and soybean oil, but almost no phase advance was induced by gelatinized cornstarch, high-amylose cornstarch, glucose, sucrose, or casein. A combination of glucose and casein without oil, vitamin, or fiber caused a significant phase advance. When cornstarch in AIN-93M was substituted with glucose, sucrose, fructose, polydextrose, high-amylose cornstarch, or gelatinized cornstarch, the amplitude of phase advance paralleled the increase in blood glucose concentration. CONCLUSIONS: Our results strongly suggest the following: (1) balanced diets containing carbohydrates/sugars and proteins are good for restricted feeding-induced entrainment of the peripheral circadian clock and (2) a balanced diet that increases blood glucose, but not by sugar alone, is suitable for entrainment. These findings may assist in the development of dietary recommendations for on-board meals served to air travelers and shift workers to reduce jet lag-like symptoms.

    DOI PubMed

  • Food anticipation in Bmal1-/- and AAV-Bmal1 rescued mice: A reply to Fuller et al

    Ralph E. Mistlberger, Ruud M. Buijs, Etienne Challet, Carolina Escobar, Glenn J. Landry, Andries Kalsbeek, Paul Pevet, Shigenobu Shibata

    Journal of Circadian Rhythms   7   11  2009.08  [Refereed]

     View Summary

    Evidence that circadian food-anticipatory activity and temperature rhythms are absent in Bmal1 knockout mice and rescued by restoration of Bmal1 expression selectively in the dorsomedial hypothalamus was published in 2008 by Fuller et al and critiqued in 2009 by Mistlberger et al. Fuller et al have responded to the critique with new information. Here we update our critique in the light of this new information. We also identify and correct factual and conceptual errors in the Fuller et al response. We conclude that the original results of Fuller et al remain inconclusive and fail to clarify the role of Bmal1 or the dorsomedial hypothalamus in the generation of food-entrainable rhythms in mice. © 2009 Mistlberger et al
    licensee BioMed Central Ltd.

    DOI PubMed

  • The dorsomedial hypothalamic nucleus is not necessary for food-anticipatory circadian rhythms of behavior, temperature or clock gene expression in mice

    Takahiro Moriya, Reiko Aida, Takashi Kudo, Masashi Akiyama, Masao Doi, Naomi Hayasaka, Norimichi Nakahata, Ralph Mistlberger, Hitoshi Okamura, Shigenobu Shibata

    EUROPEAN JOURNAL OF NEUROSCIENCE   29 ( 7 ) 1447 - 1460  2009.04  [Refereed]

     View Summary

    Circadian rhythms in mammals are regulated by a light-entrainable circadian pacemaker in the hypothalamic suprachiasmatic nucleus and food-entrainable oscillators located elsewhere in the brain and body. The dorsomedial hypothalamic nucleus (DMH) has been proposed to be the site of oscillators driving food-anticipatory circadian rhythms, but this is controversial. To further evaluate this hypothesis, we measured clock gene, temperature and activity rhythms in intact and DMH-ablated mice. A single 4-h midday feeding after an overnight fast induced mPer1 and mPer2 mRNA expression in the DMH, arcuate nucleus, nucleus of the solitary tract and area postrema, and reset daily rhythms of mPer1, mPer2 and mBMAL1 in the DMH, arcuate and neocortex. These rhythms persisted during 2 days of food deprivation after 12 days of scheduled daytime feeding. Acute induction of DMH mPer1 and mPer2 was N-methyl-d-aspartate (NMDA) receptor-dependent, whereas rhythmic expression after 6 days of restricted feeding was not. Thermal DMH lesions did not affect acute induction or rhythmic expression of clock genes in other brain regions in response to scheduled daytime feeding. DMH lesions attenuated mean daily activity levels and nocturnality but did not affect food-anticipatory rhythms of activity and body temperature in either light-dark or constant darkness. These results confirm that the DMH and other brain regions express circadian clock gene rhythms sensitive to daytime feeding schedules, but do not support the hypothesis that DMH oscillations drive food-anticipatory behavioral or temperature rhythms.

    DOI PubMed

  • Effect of chronic ethanol exposure on the liver of Clock-mutant mice

    Takashi Kudo, Toru Tamagawa, Shigenobu Shibata

    Journal of Circadian Rhythms   7   4  2009.04  [Refereed]

     View Summary

    In humans, chronic ethanol consumption leads to a characteristic set of changes to the metabolism of lipids in the liver that is referred to as an "alcoholic fatty liver (AFL)". In severe cases, these metabolic changes result in the enlargement and fibrillization of the liver and are considered risk factors for cirrhosis and liver cancer. Clock-mutant mice have been shown to display abnormal lipid metabolism and alcohol preferences. To further understand the potential interactions between ethanol consumption, lipid metabolism, and the circadian clock, we investigated the effect of chronic ethanol intake on the lipid metabolism of Clock-mutant mice. We found that ethanol treatment produced a number of changes in the liver of Clock-mutant mice without impacting the wild-type controls. First, we found that 8 weeks of exposure to ethanol in the drinking water increased the weight of the liver in Clock-mutant mice. Ethanol treatment also increased triglyceride content of liver in Clock-mutant and wild-type mice. This increase was larger in the mutant mice. Finally, ethanol treatment altered the expression of a number of genes related to lipid metabolism in the Clock-mutant mice. Interestingly, this treatment did not impact circadian clock gene expression in the liver of either genotype. Thus, ethanol produces a number of changes in the liver of Clock-mutant mice that are not seen in the wild-type mice. These changes are consistent with the possibility that disturbance of circadian rhythmicity associated with the Clock mutation could be a risk factor for the development of an alcoholic fatty liver. © 2009 Kudo et al
    licensee BioMed Central Ltd.

    DOI PubMed

  • The dorsomedial hypothalamic nucleus is not necessary for food-anticipatory circadian rhythms of behavior, temperature or clock gene expression in mice

    Takahiro Moriya, Reiko Aida, Takashi Kudo, Masashi Akiyama, Masao Doi, Naomi Hayasaka, Norimichi Nakahata, Ralph Mistlberger, Hitoshi Okamura, Shigenobu Shibata

    EUROPEAN JOURNAL OF NEUROSCIENCE   29 ( 7 ) 1447 - 1460  2009.04

     View Summary

    Circadian rhythms in mammals are regulated by a light-entrainable circadian pacemaker in the hypothalamic suprachiasmatic nucleus and food-entrainable oscillators located elsewhere in the brain and body. The dorsomedial hypothalamic nucleus (DMH) has been proposed to be the site of oscillators driving food-anticipatory circadian rhythms, but this is controversial. To further evaluate this hypothesis, we measured clock gene, temperature and activity rhythms in intact and DMH-ablated mice. A single 4-h midday feeding after an overnight fast induced mPer1 and mPer2 mRNA expression in the DMH, arcuate nucleus, nucleus of the solitary tract and area postrema, and reset daily rhythms of mPer1, mPer2 and mBMAL1 in the DMH, arcuate and neocortex. These rhythms persisted during 2 days of food deprivation after 12 days of scheduled daytime feeding. Acute induction of DMH mPer1 and mPer2 was N-methyl-d-aspartate (NMDA) receptor-dependent, whereas rhythmic expression after 6 days of restricted feeding was not. Thermal DMH lesions did not affect acute induction or rhythmic expression of clock genes in other brain regions in response to scheduled daytime feeding. DMH lesions attenuated mean daily activity levels and nocturnality but did not affect food-anticipatory rhythms of activity and body temperature in either light-dark or constant darkness. These results confirm that the DMH and other brain regions express circadian clock gene rhythms sensitive to daytime feeding schedules, but do not support the hypothesis that DMH oscillations drive food-anticipatory behavioral or temperature rhythms.

    DOI

  • Standards of evidence in chronobiology: Critical review of a report that restoration of Bmal1 expression in the dorsomedial hypothalamus is sufficient to restore circadian food anticipatory rhythms in Bmal1-/- mice

    Ralph E. Mistlberger, Ruud M. Buijs, Etienne Challet, Carolina Escobar, Glenn J. Landry, Andries Kalsbeek, Paul Pevet, Shigenobu Shibata

    Journal of Circadian Rhythms   7   3  2009.03  [Refereed]

     View Summary

    Daily feeding schedules generate food anticipatory rhythms of behavior and physiology that exhibit canonical properties of circadian clock control. The molecular mechanisms and location of food-entrainable circadian oscillators hypothesized to control food anticipatory rhythms are unknown. In 2008, Fuller et al reported that food-entrainable circadian rhythms are absent in mice bearing a null mutation of the circadian clock gene Bmal1 and that these rhythms can be rescued by virally-mediated restoration of Bmal1 expression in the dorsomedial nucleus of the hypothalamus (DMH) but not in the suprachiasmatic nucleus (site of the master light-entrainable circadian pacemaker). These results, taken together with controversial DMH lesion results published by the same laboratory, appear to establish the DMH as the site of a Bmal1-dependent circadian mechanism necessary and sufficient for food anticipatory rhythms. However, careful examination of the manuscript reveals numerous weaknesses in the evidence as presented. These problems are grouped as follows and elaborated in detail: 1. data management issues (apparent misalignments of plotted data), 2. failure of evidence to support the major conclusions, and 3. missing data and methodological details. The Fuller et al results are therefore considered inconclusive, and fail to clarify the role of either the DMH or Bmal1 in the expression of food-entrainable circadian rhythms in rodents. © 2009 Mistlberger et al
    licensee BioMed Central Ltd.

    DOI PubMed

  • ANXIOLYTIC AND ANTIDEPRESSANT EFFECTS OF ESTROGEN THROUGH ESTROGEN RECEPTOR alpha, BUT NOT beta, IN POSTPARTUM RATS AFTER WEANING PUPS

    Furuta Miyako, Ninomiya Midori, Chiba Shuichi, Kajiyama Yu, Shibata Shigenobu, Akema Tatsuo, Kunugi Hiroshi

    JOURNAL OF PHYSIOLOGICAL SCIENCES   59   333  2009  [Refereed]

  • Estrogen receptor alpha is involved in anxiety- and depressive-like behaviors in postpartum rats after weaning pups

    Furuta Miyako, Ninomiya Midori, Chiba Shuichi, Kajiyama Yu, Shibata Shigenobu, Akema Tatsuo, Kunugi Hiroshi

    NEUROSCIENCE RESEARCH   65   S221  2009  [Refereed]

    DOI

  • 時間栄養学入門・夜遅い夕食と健康のQ&A

    柴田重信

    栄養と料理 2008年12月号   74 ( 12 ) 81 - 86  2008.12

  • “体内時計の乱れ”がお腹に脂肪がつく原因だった

    柴田重信

    日経Health 2008年8月号    2008.08

  • 時間栄養における朝食の大切さ

    柴田重信

    Kellogg's Update   ( 97 ) 1 - 4  2008.04

  • セミナー 体内時計と健康(3)

    柴田重信

    臨床栄養   112 ( 3 )  2008.03

  • Gonadectomy reveals sex differences in circadian rhythms and suprachiasmatic nucleus androgen receptors in mice

    Eiko Iwahana, Ilia Karatsoreos, Shigenobu Shibata, Rae Silver

    HORMONES AND BEHAVIOR   53 ( 3 ) 422 - 430  2008.03  [Refereed]

     View Summary

    In mammals, it is well established that circadian rhythms in physiology and behavior, including the rhythmic secretion of hormones, are regulated by a brain clock located in the suprachiasmatic nucleus (SCN) of the hypothalamus. While SCN regulation of gonadal hormone secretion has been amply studied, the mechanisms whereby steroid hormones affect circadian functions are less well known. This is surprising considering substantial evidence that sex hormones affect many aspects of circadian responses, and that there are significant sex differences in rhythmicity. Our previous finding that "core" and "shell" regions of the SCN differ in their expression of clock genes prompted us to examine the possibility that steroid receptors are localized to a specific compartment of the brain clock, with the discovery that the androgen receptor (AR) is concentrated in the SCN core in mate mice. In the present study, we compare AR expression in female and male mice using Western blots and immunochemistry. Both of these methods indicate that ARs are more highly expressed in males than in females; gonadectomy eliminates and androgen treatment restores these sex differences. At the behavioral level, gonadectomy produces a dramatic loss of the evening activity onset bout in males, but has no such effect in females. Treatment with testosterone, or with the non-aromatizable androgen dihydrotestosterone, restores male locomotor activity and eliminates sex differences in the behavioral response. The results indicate that androgenic hormones regulate circadian responses, and suggest an SCN site of action. (c) 2007 Elsevier Inc. All rights reserved.

    DOI PubMed

  • セミナー 体内時計と健康(2)

    柴田重信

    臨床栄養   112 ( 2 )  2008.02

  • 体内時計と食

    柴田重信

    キューピーニュース   ( 406 )  2008

  • 生命現象に潜むリズムを解明

    柴田重信

    WIN WING(アズワン株式会社)    2008

  • 体内時計と食

    柴田重信

    FFIジャーナル(食品・食品添加物研究誌)   213 ( 1 )  2008

  • 時刻・時間における体内時計機構の役割

    柴田重信

    動物心理学研究   58 ( 1 ) 21 - 31  2008

    DOI

  • セミナー 体内時計と健康(1)

    柴田重信

    臨床栄養   112 ( 1 )  2008.01

  • 睡眠覚醒のリズム

    守屋孝洋, 柴田重信

    小児内科   40 ( 1 )  2008.01

  • Clock mutation facilitates accumulation of cholesterol in the liver of mice fed a cholesterol and/or cholic acid diet

    Takashi Kudo, Mihoko Kawashima, Toru Tamagawa, Shigenobu Shibata

    AMERICAN JOURNAL OF PHYSIOLOGY-ENDOCRINOLOGY AND METABOLISM   294 ( 1 ) E120 - E130  2008.01  [Refereed]

     View Summary

    Cholesterol (CH) homeostasis in the liver is regulated by enzymes of CH synthesis such as 3-hydroxy-3-methylglutaryl coenzyme A reductase (HMGCR) and catabolic enzymes such as cytochrome P-450, family 7, subfamily A, and polypeptide 1 (CYP7A1). Since a circadian clock controls the gene expression of these enzymes, these genes exhibit circadian rhythm in the liver. In this study, we examined the relationship between a diet containing CH and/or cholic acid (CA) and the circadian regulation of Hmgcr, low-density lipoprotein receptor (Ldlr), and Cyp7a1 gene expression in the mouse liver. A 4-wk CA diet lowered and eventually abolished the circadian expression of these genes. Not only clock genes such as period homolog 2 (Drosophila) (Per2) and brain and muscle arnt-like protein-1 (Bmal1) but also clock-controlled genes such as Hmgcr, Ldlr, and Cyp7a1 showed a reduced and arrhythmic expression pattern in the liver of Clock mutant mice. The reduced gene expression of Cyp7a1 in mice fed a diet containing CA or CH + CA was remarkable in the liver of Clock mutants compared with wild-type mice, and high liver CH accumulation was apparent in Clock mutant mice. In contrast, a CH diet without CA only elevated Cyp7a1 expression in both wild-type and Clock mutant mice. The present findings indicate that normal circadian clock function is important for the regulation of CH homeostasis in the mouse liver, especially in conjunction with a diet containing high CH and CA.

    DOI PubMed

  • Multifactorial regulation of daily rhythms in expression of the metabolically responsive gene Spot14 in the mouse liver

    Akinori Ishihara, Eriko Matsumoto, Kazumasa Horikawa, Takashi Kudo, Eiko Sakao, Ayako Nemoto, Katsuro Iwase, Hajime Sugiyama, Yutaka Tamura, Shigenobu Shibata, Masaki Takiguchi

    JOURNAL OF BIOLOGICAL RHYTHMS   22 ( 4 ) 324 - 334  2007.08  [Refereed]

     View Summary

    Spot14 is a putative transcriptional regulator for genes involved in fatty acid synthesis. The Spot14 gene is activated in response to lipogenic stimuli such as dietary carbohydrate and is also under circadian regulation. The author's investigated factors responsible for daily oscillation of Spot14 expression. If mice were kept under a 12-h light/ 12-h dark cycle with ad libitum feeding, Spot14 mRNA levels in the liver reached a peak at an early dark period when mice, as nocturnal animals, start feeding. Under fasting, while Spot14 mRNA levels were generally decreased, the rhythmicity was still maintained, suggesting contribution of both nutritional elements and circadian clock factors on robust rhythmicity of Spot14 expression. Effects of circadian clock factors were confirmed by the observations that the circadian rhythm of Spot14 expression was seen also under the constant darkness and that the rhythmicity was lost in Clock mutant mice. When mice were housed in short-photoperiod (6-h light/18-h dark) and long-photoperiod (18-h light/6-h dark) cycles, rhythms of Spot14 mRNA levels were phase advanced and phase delayed, respectively, being concordant with the notion that Spot14 expression is under the control of the light-entrainable oscillator. As for nutritional mediators, in the liver of db/db mice exhibiting hyperinsulinemia-accompanied hyperglycemia, Spot14 mRNA levels were constantly high without apparent rhythmicity, consistent with previous observations for strong activation of the Spot14 gene by glucose and insulin. Restricted feeding during the 4-h mid-light period caused a phase advance of the Spot14 expression rhythm. On the other hand, restricted feeding during the 4-h mid-dark period led to damping of the rhythmicity, apparently resulting from the separation of phases between effects of the light/dark cycle and feeding on Spot14 expression. Thus, the daily rhythm of Spot14 expression in the liver is under the control of the light-entrainable oscillator, food-entrainable oscillator, and food-derived nutrients, in a separate or cooperative manner.

    DOI PubMed

  • Attenuating effect of Clock mutation on triglyceride contents in the ICR mouse liver under a high-fat diet

    Takashi Kudo, Toru Tarnagawa, Mihoko Kawashima, Natsuko Mito, Shigenobu Shibata

    JOURNAL OF BIOLOGICAL RHYTHMS   22 ( 4 ) 312 - 323  2007.08  [Refereed]

     View Summary

    Energy homeostasis is subjected to a circadian control that synchronizes energy intake and expenditure. The transcription factor CLOCK, a key component of the molecular circadian clock, controls many kinds of rhythms, such as those for locomotor activity, body temperature, and metabolic functions. The purpose of the present study is to understand the function of the Clock gene during lipid metabolism in the liver using Clock-mutant mice. Clock-mutant mice with an ICR background were fed a high-fat diet for 13 weeks, and liver triglyceride, serum triglyceride, and serum free fatty acid levels were examined. Triglyceride content in the liver was significantly less increased in Clock-mutant mice on a high-fat diet compared to wild-type mice on a high-fat diet. Acsl4 and Fabp1 mRNA levels in the liver showed daily rhythms in wild-type mice, In contrast, Clock-mutant mice had attenuated daily rhythms of Acs14 and Fabp1 gene expression in the liver under both normal and high-fat diet conditions compared to wild-type mice. In Clock-mutant mice, suppression of Acs14 and Fabp1 mRNA in the liver under high-fat diet conditions may have attenuated the accumulation of triglycerides in the liver compared to wild-type mice under the same conditions. In conclusion, the authors demonstrate that mice with a Clock mutation showed less triglyceride accumulation in the liver through the suppression of Acs14 and Fabp1 gene expression when fed a high-fat diet compared to wild-type mice fed the same diet.

    DOI PubMed

  • Optimization of dosing schedule of daily inhalant dexamethasone to minimize phase shifting of clock gene expression rhythm in the lungs of the asthma mouse model

    Naomi Hayasaka, Tsuyoshi Yaita, Tomoyuki Kuwaki, Sato Honma, Ken-ichi Honma, Takashi Kudo, Shigenobu Shibata

    ENDOCRINOLOGY   148 ( 7 ) 3316 - 3326  2007.07  [Refereed]

     View Summary

    Glucocorticoid receptor agonists such as dexamethasone ( DEXA) have been recommended for the treatment of asthma. An increased frequency of dosing with these drugs seems preferable for cases of severe or uncontrolled asthma. The purpose of this experiment was to find the appropriate dosing schedule ( frequency and timing) for DEXA inhalation based on chronotherapeutic dosing to minimize phase shifts of clock function in the lungs of the ovalbumin-treated asthmatic mouse. The daily rhythm of clock gene expression was similar between control and ovalbumin-treated mice. Acute inhalation of DEXA significantly increased mPer1 gene expression in the lungs but not the liver of mice. Daily exposure of DEXA at zeitgeber time 0 ( lights on) or at zeitgeber time 18 ( 6 h after lights off) for 6 d caused a phase advance or phase delay of bioluminescence rhythm in the lungs, respectively, similar to light-induced phase shifts in locomotor activity rhythm. Daily zeitgeber time 0 exposure to DEXA attenuated the expression level of the mClca3 gene, which is associated with mucus overproduction, and there was a phase-advancing peak time of the mClca3 rhythm. The present results denote the importance of selecting the most appropriate time of day for nebulizer administration of DEXA to minimize adverse effects such as the phase shifting of clock function in asthmatic lungs. This is the first report of a successful protocol that could obtain phase shifts of clock gene expression rhythm in isolated peripheral organs in vivo.

    DOI PubMed

  • PPAR alpha is a potential therapeutic target of drugs to treat circadian rhythm sleep disorders

    Hidenori Shirai, Katsutaka Oishi, Takashi Kudo, Shigenobu Shibata, Norio Ishida

    BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS   357 ( 3 ) 679 - 682  2007.06  [Refereed]

     View Summary

    Recent progress at the molecular level has revealed that nuclear receptors play an important role in the generation of mammalian circadian rhythms. To examine whether peroxisome proliferator-activated receptor alpha (PPAR alpha) is involved in the regulation of circadian behavioral rhythms in mammals, we evaluated the locomotor activity of mice administered with the hypolipidemic PPAR alpha ligand, bezafibrate. Circadian locomotor activity was phase-advanced about 3 h in mice given bezafibrate under light-dark (LD) conditions. Transfer from LD to constant darkness did not change the onset of activity in these mice, suggesting that bezafibrate advanced the phase of the endogenous clock. Surprisingly, bezafibrate also advanced the phase in mice with lesions of the suprachiasmatic nucleus (SCN; the central clock in mammals). The circadian expression of clock genes such as period2, BMAL1, and Rev-erba was also phase-advanced in various tissues (cortex, liver, and fat) without affecting the SCN. Bezafibrate also phase-advanced the activity phase that is delayed in model mice with delayed sleep phase syndrome (DSPS) due to a Clock gene mutation. Our results indicated that PPARa is involved in circadian clock control independently of the SCN and that PPARa could be a potent target of drugs to treat circadian rhythm sleep disorders including DSPS. (c) 2007 Elsevier Inc. All rights reserved.

    DOI PubMed

  • 朝食は体内時計にどのように影響するか?

    柴田重信

    栄養と料理 2007年4月号    2007.04

  • Constant light housing during nursing causes human DSPS (delayed sleep phase syndrome) behaviour in Clock-mutant mice

    Yukako Wakatsuki, Takashi Kudo, Shigenobu Shibata

    EUROPEAN JOURNAL OF NEUROSCIENCE   25 ( 8 ) 2413 - 2424  2007.04  [Refereed]

     View Summary

    Delayed sleep phase syndrome (DSPS) is very often seen among patients with sleep-wake rhythm disorders. Humans with the 3111C allele of the human Clock gene tend to demonstrate a higher evening preference on the morningness-eveningness (ME) preference test. DSPS is thought to be an extreme form of this evening preference. Clock-mutant mice have been proposed as an animal model of evening preference. In this study, we looked at whether constant light (LL) housing of Clock-mutant mice during lactation would result in evening preference and/or DSPS. Housed under light-dark (LD) or constant dark (DD) conditions during the lactation period, both wild-type and Clock-mutant mice did not show a phase-delay in the locomotor activity measured under light-dark conditions, whereas constant light housing during lactation significantly caused a delayed onset. The magnitude of the delay during the light-dark cycle was positively associated with free-running period measured during constant darkness. Among wild, heterozygote, and homozygote pups born from heterozygous dams, only homozygote pups showed a delayed onset. Constant light-housed Clock-mutant mice exhibited a lower number and delayed peak of phospho-MAPK-immunoreactive cells in core regions of the suprachiasmatic nucleus (SCN) compared to light-dark housed wild-type or Clock-mutant mice. Activity onset returned to normal with daily melatonin injection at the lights-off time for 5 days. The present results demonstrate that Clock-mutant mice exposed to constant light during lactation can function as an animal model of DSPS and can be used to gain an understanding of the ethological aspects of DSPS as well as to find medication for its treatment.

    DOI PubMed

  • ORL1 receptor-mediated down-regulation of mPER2 in the suprachiasmatic nucleus accelerates re-entrainment of the circadian clock following a shift in the environmental light/dark cycle

    Kazuko Miyakawa, Ayumi Uchida, Tomomi Shiraki, Koji Teshima, Hiroshi Takeshima, Shigenobu Shibata

    NEUROPHARMACOLOGY   52 ( 3 ) 1055 - 1064  2007.03  [Refereed]

     View Summary

    The circadian pacemaker in the suprachiasmatic nucleus (SCN) generates the near 24-h period of the circadian rhythm and is entrained to the 24-h daily cycle by periodic environmental signals, such as the light/dark cycle (photic signal), and can be modulated by various drugs (non-photic signals). The mechanisms by which non-photic signals modulate the circadian clock are not well understood in mice. In mice, many reportedly non-photic stimuli have little effect on the circadian rhythm in vivo. Herein, we investigated the molecular mechanism in W-212393-induced phase advance using mice. W-212393 caused a significant phase advance of locomotor activity rhythm in mice at subjective day. Injection of W-212393 during subjective day elicited down-regulation of mPER2 protein in the SCN shell region, but not mPei-2 mRNA. Administration of W-212393 during subjective day failed to produce phase advance in mPet-2-mutant mice as well as in ORL1 receptor deficient mice. Furthermore, we show that such inhibition of mPER2 accelerates re-entrainment of the circadian clock following an abrupt shift in the environmental light/dark cycle, such as occurs with transmeridian flight. The present results suggest that post-translational down-regulation of mPER2 protein in the shell region of mouse SCN may be involved in W-212393-induced non-photic phase advance. (c) 2006 Elsevier Ltd. All rights reserved.

    DOI PubMed

  • Differential effect of lithium on the circadian oscillator in young and old hamsters

    Eiko Iwahana, Toshiyuki Hamada, Ayumi Uchida, Shigenobu Shibata

    BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS   354 ( 3 ) 752 - 756  2007.03  [Refereed]

     View Summary

    Lithium is one of the most commonly used drugs in the prophylaxis and treatment of bipolar disorder. It is also known to lengthen circadian period in several organisms. Previously, we reported that there was the association between lengthening circadian period by lithium and GSK-3 protein and its enzyme activity in the mouse suprachiasmatic nucleus (SCN). In this study, we show that lithium affects the circadian oscillator in young and old hamster SCN, in an age-dependent manner. We found that basal levels of phosphorylated GSK-3 (pGSK-3) protein expression in old hamsters are much lower than that in young hamsters. Furthermore, in the old hamsters, lithium did not affect the period of the locomotor activity rhythm or pGSK-3 expression, while changing period and pGSK-3 in the younger animals. These results indicate that the content of pGSK-3 in the SCN has an important role in age-dependent effects of lithium on the circadian oscillator. (c) 2007 Elsevier Inc. All rights reserved.

    DOI PubMed

  • Circadian rhythms in the CNS and peripheral clock disorders: The cireadian clock and hyperlipidemia

    Takashi Kudo, Kazumasa Horikawa, Shigenobu Shibata

    JOURNAL OF PHARMACOLOGICAL SCIENCES   103 ( 2 ) 139 - 143  2007.02  [Refereed]

     View Summary

    A circadian clock controls various physiological and behavioral rhythms. In mammals, a master circadian clock exists in the suprachiasmatic nucleus of the hypothalamus, and slave oscillators can be found in most tissues. These circadian oscillations are controlled by "clock genes". The negative feedback loop is thought to function as a molecular mechanism of the circadian clock. It is plausible that clock genes may control lipid metabolism through so-called clock-controlled genes and that lipid metabolism-related clock-controlled genes may play important roles in the circadian change of lipid metabolism. Recently research has focused on the relationships between the clock system and lipid metabolism. In this review, we discuss the following items: 1) circadian clock system, 2) effect of the diet on clock gene expression, 3) effect of clock mutation on lipid metabolism, and 4) effect of streptozotocin-induced diabetes and ob mutation on clock gene expression and lipid metabolism. In this review we have summarized how the circadian clock affects lipid metabolism through the expression of lipid metabolism-related clock-controlled genes and at the same time discussed how abnormal metabolism of lipid affects the expression of clock genes. Further experiments are needed to elucidate the detailed mechanism of interaction between clock genes and lipid metabolisms.

    DOI PubMed

  • Circadian rhythms in the CNS and peripheral clock disorders: Preface

    Shigenobu Shibata

    JOURNAL OF PHARMACOLOGICAL SCIENCES   103 ( 2 ) 133 - 133  2007.02  [Refereed]

    DOI PubMed

  • 朝日でリセット心と体「太陽の光をたっぷり浴びることで、体内時計を調節」

    柴田重信

    心の健康ニュース   ( 312 )  2006.09

  • [Chronopharmacology].

    Shibata S

    Nihon yakurigaku zasshi. Folia pharmacologica Japonica   128 ( 2 ) 112 - 114  2006.08  [Refereed]

    PubMed

  • Circadian Clock mutation in dams disrupts nursing behavior and growth of pups

    K Hoshino, Y Wakatsuki, M Iigo, S Shibata

    ENDOCRINOLOGY   147 ( 4 ) 1916 - 1923  2006.04

     View Summary

    To understand the role of the circadian molecular clock in mouse reproduction, we investigated the daily rhythms associated with nursing and pup growth in Clock-mutant mice maintained under light-dark housing conditions. The daily rhythm associated with the maternal behavior of crouching had a strong diurnal peak and two weak nocturnal peaks in wild-type dams, whereas homozygotes (Clock/Clock) exhibited no significant peaks in activity. Wild-type, but not Clock-mutant, dams showed high, rhythmic levels of prolactin content in serum that corresponded with crouching. Pup body weight increased at a significantly slower rate in Clock-mutant dams compared with wild-type dams under all experimental conditions when the pups ranged from 10-15 in number. Heterozygote dams equally bred wild-type, heterozygote, or homozygote pups. The amount of milk secreted from dams, as calculated by the increase in pup body weight through suckling, was lower in Clock-mutant mothers vs. wild-type mice. When Clock-mutant dams gave birth to more than 10 pups, survival was poor for offspring until the time of weaning. The present results demonstrate that Clock mutation disrupts daily maternal behavior and the growth and survival rate of pups, especially with the breeding of more than 10 pups.

    DOI

  • 肝臓時計遺伝子発現系を利用した食品添加物の効能・副作用評価

    柴田重信

    第12回研究成果報告書(日本食品化学研究振興財団)    2006

  • A role of nociceptin in the entrainment mechanism of the biological clock

    Koji Teshima, Shigenobu Shibata

    Japanese Journal of Neuropsychopharmacology   25 ( 5 ) 203 - 211  2005.10

     View Summary

    The ORL1 receptor (nociceptin receptor), the fourth member of the opioid receptor family, is widely distributed throughout the central nervous system and mediates diverse physiological functions including pain, emotion, learning and memory. Nociceptin receptors are densely expressed in the suprachiasmatic nucleus, the principal pacemaker of circadian rhythms, and nociceptin inhibits light-induced phase advances (Allen et al, 1999)
    however, nociceptin's role in nonphotic entrainment has not been described. We used a small-molecule nociceptin receptor agonist capable of crossing the brain blood-brain barrier, and demonstrated that nociceptin acts as a nonphotic zeitgeiber. In this review, we describe a role for nociceptin in the biological clock and discuss the possibility of developing nociceptin receptor agonists for treatment of circadian rhythm disorders.

    PubMed

  • Circadian rhythm study from anticipatory behavior to drug treatment

    Shigenobu Shibata

    Japanese Journal of Neuropsychopharmacology   25 ( 5 ) 251 - 258  2005.10

     View Summary

    Precise, rhythmic, daily change of the internal milieu is a conspicuous feature of all living organisms. It affects temporal patterns of all kinds of behaviors during a day and deeply influences both the social structure and daily life of individual human beings. These daily variations arise from the internal circadian mechanisms. Three functions of the endogenous clock are discriminated as rhythm generation, entrainment to light-dark cycle and output from the clock. The endogenous clock is localized in the suprachiasmatic nucleus (SCN) in mammals. Recent papers demonstrated strong expression of clock genes such as Per1, Per2 and Per3 in the SCN. Circadian oscillation is basically regulated by the transcription/translation feedback system of the Per gene in mammals. As serotonin/antidepressant and GABA/benzodiazepine drugs affect the light and non-light-induced entrainment, these drugs can regulate the circadian oscillation of clock genes and environmental stimuli-induced change of Per gene expression in the SCN. There are two main stimuli that entrain circadian rhythm, the light-dark cycle (LD) and restricted feeding. Light resets the circadian clock with induction of Per1 and Per2 gene in the SCN, the locus of a main oscillator. Mice were allowed access to food for 4 h during daytime (7 h in advance of feeding time) under LD or constant darkness. The peaks of mPer1 and mPer2 mRNA in the cerebral cortex and liver were advanced 6-12 h after 6 days of RF, whereas those in SCN were unaffected. The increase of mPer expression by RF treatment was observed in SCN-lesioned mice. The present results suggest that RF strongly entrained the expression of mPer and clock-controlled genes in the cerebral cortex and liver without affecting light-dependent SCN clock function.

    PubMed

  • Nonphotic entrainment of the circadian body temperature rhythm by the selective ORL1 receptor agonist W-212393 in rats

    K Teshima, M Minoguchi, S Tounai, A Ashimori, J Eguchi, CN Allen, S Shibata

    BRITISH JOURNAL OF PHARMACOLOGY   146 ( 1 ) 33 - 40  2005.09

     View Summary

    1 We synthesized a small-molecule opioid receptor-like 1 (ORL1) receptor agonist, 2-{3-[1-((1R)-acenaphthen- 1-yl)piperidin-4-yl]-2,3-dihydro-2-oxo-benzimidazol-1-yl}-N-methylacetamide (W-212393), and investigated its effect on the circadian body temperature rhythm of rats.
    2 W-212393 has high affinity for ORL1 receptors in the rat cerebral cortex and human ORL1 receptors expressed in HEK293 cells with K-i values of 0.76 and 0.50 nM, respectively.
    3 W-212393 concentration-dependently stimulated GTP gamma 35S binding and its efficacy was similar to nociceptin/orphanin FQ (N/OFQ), suggesting that W-212393 is a full agonist at ORL1 receptors.
    4 W-212393 dose-dependently occupied ORL1 receptors following intraventricular or intraperitoneal administration, suggesting that W-212393 is a brain-penetrating compound.
    5 W-212393 (100 nM) and N/OFQ (100 nM) significantly suppressed the activity of spontaneously firing rat suprachiasmatic nucleus neurons. These suppressive effects were blocked by an ORL1 receptor antagonist, J-113397 (1 mu M).
    6 W-212393 (3 mg kg(-1), i.p.) induced a significant phase advance at circadian time 6 (CT6) and CT9, but not at other CTs. The magnitude of the W-212393 (0.3-3 mg kg(-1), i.p.)-induced phase advance was dose-dependent and greater than those produced by 8-hydroxy-2-(di-n-propylamino)tetralin (0.3-3 mg kg(-1), i.p.) or melatonin (0.3-3 mg kg(-1), i.p.). The W-212393 (3 mg kg(-1), i.p.)-induced phase advance was antagonized by J-113397 (10 mg kg(-1), i.p.).
    7 W-212393 (3 mg kg(-1), i.p.) significantly accelerated the re-entrainment of the body temperature rhythm to a 6 h advanced light-dark cycle.
    8 These results indicate that activation of ORL1 receptors contributes to the circadian entrainment and W-212393 may represent an interesting agent for the study of circadian rhythms.

    DOI

  • Altered food-anticipatory activity rhythm in Cryptochrome-deficient mice

    M Iijima, S Yamaguchi, GTJ van der Horst, Bonnefont, X, H Okamura, S Shibata

    NEUROSCIENCE RESEARCH   52 ( 2 ) 166 - 173  2005.06

     View Summary

    In nocturnal rodents, restricted feeding to daytime (RF) causes feeding-associated diurnal locomotor activity that persists for the next 1-2 days when food is withheld. Along with this anticipatory behavior, the expression pattern of clock genes such as mPer1 and mPer2 changes from a nocturnal to diurnal pattern in the liver and cerebral cortex but not in the suprachiasmatic nucleus (SCN). Whether the molecular clockwork, in which mCry1 and mCry2 genes are essential components, is involved in food-anticipatory circadian rhythms is unknown.
    In this study, we investigated the impact of the absence of mCRY products upon the locomotion pattern induced by RF. RF caused an increase in daytime activity that lasted even for 2 days after food was withheld, in wild-type and mCry1-1-mCry2-1-mice. However, RF-induced activity was less stable and appeared more gradually in mutant mice. Similar results were obtained with mice housed under constant darkness or with SCN-lesioned wild-type and mutant mice. Our data reveal that mCry proteins are basically dispensable for food-entrainable oscillation. However, it is also important to note that mCry deficiency affects the stability and development of RF-induced anticipatory locomotor activity. (c) 2005 Elsevier Ireland Ltd and the Japan Neuroscience Society. All rights reserved.

    DOI

  • Inhibitory action of 5-HT1A agonist MKC-242 on triazolam-induced phase advances in hamster circadian activity rhythms

    S Yokota, T Moriya, S Shibata

    JOURNAL OF PHARMACOLOGICAL SCIENCES   98 ( 1 ) 103 - 106  2005.05

     View Summary

    Mammalian circadian rhythms can be entrained by photic and non-photic stimuli. Although we know that non-photic entrainment interferes with photic entrainment signals, there is no information about whether photic entrainment interferes with non-photic entraining signals. We examined whether triazolam (TRZ), a non-photic form of entrainment, could be attenuated by pre-treatment with (S)-5-[3-[(1,4-benzodioxan-2-ylmethyl)amino]propoxy]-1,3-benzodioxole hydrochloride (MKC-242), a photic enhancing drug. We found that TRZ-induced phase advances in hamster behavioral circadian rhythms and the increase of locomotor activity were both significantly attenuated by MKC-242. Thus, in hamsters, the photic enhancing drug MKC-242 seemed to hinder benzodiazepine-induced non-photic entrainment.

    DOI

  • 生体リズムに関連した運動処方の開発とヒトへの応用

    柴田重信

    第7回(平成16年度)選定 宇宙環境利用に関する公募地上研究 研究成果報告書(概要版)    2005

  • Rapid damping of food-entrained circadian rhythm of clock gene expression in clock-defective peripheral tissues under fasting conditions

    K Horikawa, Y Minami, M Iijima, M Akiyama, S Shibata

    NEUROSCIENCE   134 ( 1 ) 335 - 343  2005

     View Summary

    Restricted feeding-induced free-running oscillation of clock genes in the liver was studied in homozygous Clock-mutant (Clock/Clock) mice. Similar to wild-type mice, Clock/Clock mice showed robust food-anticipatory behavioral activity in accordance with a restricted feeding schedule. Also, the peak of all clock gene mRNAs tested was phase-advanced in the liver of Clock/Clock mice as well as wild-type mice, although the amplitude of clock gene expression was low in Clock/Clock mice. The food-anticipatory behavioral rhythm in Clock/Clock mice maintained a period similar to wild-type mice during 2-day fasting after the cessation of restricted feeding. However, during the fasting days after temporal feeding cues were removed, the oscillation of clock genes in the liver and heart, excluding the suprachiasmatic nuclei, appeared to result in arrhythmicity in Clock/Clock mice. Thus, although the CLOCK-based molecular mechanism is not required for the expression of food-anticipatory activity, intact CLOCK protein might be involved in sustaining several cycles of peripheral circadian oscillations after restricted feeding-induced resetting. (c) 2005 Published by Elsevier Ltd on behalf of IBRO.

    DOI

  • Reduced food anticipatory activity in genetically orexin (hypocretin) neuron-ablated mice

    M Akiyama, T Yuasa, N Hayasaka, K Horikawa, T Sakurai, S Shibata

    EUROPEAN JOURNAL OF NEUROSCIENCE   20 ( 11 ) 3054 - 3062  2004.12

     View Summary

    Daily restricted feeding (RF) produces an anticipatory locomotor activity rhythm and entrains the peripheral molecular oscillator independently of the central pacemaker located in the suprachiasmatic nucleus (SCN). As orexins (hypocretins) are neuropeptides that coordinate sleep/wake patterns and motivated behaviours, such as food seeking, we studied the involvement of orexin in the food anticipatory activity (FAA) induced by RF. Daily RF shifted the mRNA rhythm of a clock-controlled gene mDbp in the cerebral cortex and caudate putamen but not in the SCN. Under these experimental conditions, prepro-orexin mRNA and orexin A immunoreactivity in the lateral hypothalamic area (LHA) did not show daily variation. Fasting increased the number of orexin A-ir cells, while RF did not. However, RF shifted the peak of Fos expression of the orexin neurons from night to day. Genetic ablation of orexin neurons in orexin/ataxin-3 transgenic mice severely reduced the formation of FAA under RF conditions. The expression of mNpas2 mRNA, a transcription factor thought to be involved in regulation of the food entrainable oscillator as well as mPer1 and mBmal1 mRNA, was reduced in the forebrain of orexin/ataxin-3 mice. Based on these results, we suggest that activity of the orexin neuron in the LHA contributes to the promotion and maintenance of FAA.

    DOI

  • Cholesterol diet enhances daily rhythm of Pai-1 mRNA in the mouse liver

    T Kudo, E Nakayama, S Suzuki, M Akiyama, S Shibata

    AMERICAN JOURNAL OF PHYSIOLOGY-ENDOCRINOLOGY AND METABOLISM   287 ( 4 ) E644 - E651  2004.10

     View Summary

    Myocardial infarction frequently occurs in the morning, a phenomenon in part resulting from the downregulation of fibrinolytic activity. Plasminogen activator inhibitor-1 (PAI-1) is a key factor behind fibrinolytic activity, and its gene expression is controlled under the circadian clock gene in the mouse heart and liver. Hypercholesterolemia has been associated with impaired fibrinolysis due to enhanced PAI-1 activity, which has also been implicated in atherosclerosis. The aim of this study was to decipher whether the Pai-1 gene is still expressed daily with hypercholesterolemia. Hypercholesterolemia (1% cholesterol diet) did not significantly affect the daily expression of clock genes (Per2 and Bmal1) and clock-controlled genes (Dbp and E4bp4) in the liver (P&gt;0.05); however, daily expression of the Pai-1 gene and Pai-1 promoter regulating factor genes such as Nr4a1 was significantly upregulated (P&lt;0.01). Daily restricted feeding for 4 h during the day reset the gene expression of Per2, Pai-1, Nr4a1, and Tnf-&alpha;. Lesion of the suprachiasmatic nucleus, the location of the main clock system, led to loss of Per2 and Pai-1 daily expression profiles. In the present experiments, we demonstrated that hypercholesterolemia enhanced daily expression of the Pai-1, Tnf-&alpha;, and Nr4a1 genes in the mouse liver without affecting clock and clock-controlled genes. Therefore, the risk or high frequency of acute atherothrombotic events in the morning still seems to be a factor that may be augmented under conditions of hypercholesterolemia.

    DOI

  • Phase-resetting response to (+)8-OH-DPAT, a serotonin 1A/7 receptor agonist, in the mouse in vivo

    K Horikawa, S Shibata

    NEUROSCIENCE LETTERS   368 ( 2 ) 130 - 134  2004.09

     View Summary

    Several lines of evidence indicate that 5-HT neuronal systems may play a critical role for the non-photic entrainment of the rodent circadian clock. Although it is well established that (+)8-hydroxy-2-(di-n-propylamino)tetralin [(+)8-OH-DPAT], a 5-HT1A/7 receptor agonist, causes a phase-advance of behavioral rhythm in hamsters, little is known whether this agent produces phase shifts of activity rhythm in mice. Therefore, we examined the effect of (+)8-OH-DPAT on the mouse locomotor activity rhythm. Systemic administration of this chemical at mid-subjective daytime induced a clear and dose-dependent phase advance, while there were no significant phase shifts at other times (early-subjective day, late-subjective day, or subjective night). Additionally, (+)8-OH-DPAT accelerated the re-entrainment of mouse behavioral rhythm to a 6-h advanced light-dark cycle. These results suggest that we can use mice for understanding the molecular mechanism of (+)8-OH-DPAT-induced phase shift because of availability of clock gene targeted mice. (C) 2004 Elsevier Ireland Ltd. All rights reserved.

    DOI

  • Neural regulation of the hepatic circadian rhythm

    S Shibata

    ANATOMICAL RECORD PART A-DISCOVERIES IN MOLECULAR CELLULAR AND EVOLUTIONARY BIOLOGY   280A ( 1 ) 901 - 909  2004.09

     View Summary

    A microarray analysis experiment has revealed that there are many genes, including so-called clock genes, expressing a circadian rhythm in the liver. The clock genes mentioned above are expressed not only in the suprachiasmatic nucleus (SCN) of the hypothalamus, where the master clock exists, but also in other brain regions and various peripheral tissues. In the liver, clock genes are abundantly expressed and show a clear circadian rhythm. Thus, clock genes seem to play a critical role in the molecular clockworks of both the SCN and the liver. Although oscillation of clock genes in the liver is controlled under the circadian clock mechanism in the SCN, we do not know the resetting signals on liver clock function. Over the past few years, use of the pseudorabies virus, a transsynaptic tract tracer, has allowed us to map neural connections between the SCN and peripheral tissues in several physiological systems. Communication between the SCN and peripheral tissues occurs through autonomic nervous systems involving the sympathetic and parasympathetic neurons. This review mainly describes both anatomical and physiological experiments to reveal the sympathetic control over liver clock function. Although further study is necessary to produce the precise mechanism underlying neural control of liver clock systems, evolution of this mechanism will help our understanding of liver clock functions such as drug metabolism and energy metabolism. (C) 2004 Wiley-Liss, Inc.

    DOI

  • Daily injection of insulin attenuated impairment of liver circadian clock oscillation in the streptozotocin-treated diabetic mouse

    K Kuriyama, K Sasahara, T Kudo, S Shibata

    FEBS LETTERS   572 ( 1-3 ) 206 - 210  2004.08

     View Summary

    Recent studies have shown a dampened amplitude of clock gene rhythm in the heart and liver of streptozotocin (STZ)-treated rats and mice, however it is unknown whether impairment is due to dysfunction of the suprachiasmatic nucleus (SCN) or not. Rhythmic expression of mPER2 was dampened in the STZ-treated mouse liver but not SCN and cerebral cortex. Injection of insulin could normalize an impairment of mPer2 and mPER2 expression rhythm in the liver, when it was injected at nighttime, but not at daytime. In the present study, we demonstrated that insulin-dependent diabetes impaired oscillation of the peripheral clock gene and its product. Insulin injection can recover dampened oscillation of the peripheral clock depending on its injection time. (C) 2004 Published by Elsevier B.V. on behalf of the Federation of European Biochemical Societies.

    DOI

  • Night-time restricted feeding normalises clock genes and Pai-1 gene expression in the db/db mouse liver

    T Kudo, M Akiyama, K Kuriyama, M Sudo, T Moriya, S Shibata

    DIABETOLOGIA   47 ( 8 ) 1425 - 1436  2004.08

     View Summary

    Aims/hypothesis. An increase in PAI-1 activity is thought to be a key factor underlying myocardial infarction. Mouse Pai-1 (mPai-1) activity shows a daily rhythm in vivo, and its transcription seems to be controlled not only by clock genes but also by humoral factors such as insulin and triglycerides. Thus, we investigated daily clock genes and mPai-1 mRNA expression in the liver of db/db mice exhibiting high levels of glucose, insulin and triglycerides.
    Methods. Locomotor activity was measured using an infrared detection system. RT-PCR or in situ hybridisation methods were applied to measure gene expression. Humoral factors were measured using measurement kits.
    Results. The db/db mice showed attenuated locomotor activity rhythms. The rhythmic expression of mPer2 mRNA was severely diminished and the phase of mBmal1 oscillation was advanced in the db/db mouse liver, whereas mPai-1 mRNA was highly and constitutively expressed. Night-time restricted feeding led to a recovery not only from the diminished locomotor activity, but also from the diminished Per2 and advanced mBmal1 mRNA rhythms. Expression of mPai-1 mRNA in db/db mice was reduced to levels far below normal. Pioglitazone treatment slightly normalised glucose and insulin levels, with a slight reduction in mPai-1 gene expression.
    Conclusions/interpretation. We demonstrated that Type 2 diabetes impairs the oscillation of the peripheral oscillator. Night-time restricted feeding rather than pioglitazone injection led to a recovery from the diminished locomotor activity, and altered oscillation of the peripheral clock and mPai-1 mRNA rhythm. Thus, we conclude that scheduled restricted food intake may be a useful form of treatment for diabetes.

    DOI

  • The role of Clock in the plasticity of circadian entrainment

    R Udo, T Hamada, K Horikawa, E Iwahana, K Miyakawa, K Otsuka, S Shibata

    BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS   318 ( 4 ) 893 - 898  2004.06

     View Summary

    The mammalian circadian clock lying in suprachiasmatic nucleus (SCN) is synchronized to about 24h by the environmental light-dark cycle (LD). The circadian clock exhibits limits of entrainment above and below 24h, beyond which it will not entrain. Little is known about the mechanisms regulating the limits of entrainment. In this study, we show that wild-type mice entrain to only an LD 24 h cycle, whereas Clock mutant mice can entrain to an LD 24, 28, and 32 h except for LD 20 h and LD 36 h cycle. Under an LD 28 h cycle, Clock mutant mice showed a clear rhythm in Per2 mRNA expression in the SCN and behavior. Light response was also increased. This is the first report to show that the Clock mutation makes it possible to adapt the circadian oscillator to a long period cycle and indicates that the clock gene may have an important role for the limits of entrainment of the SCN to LD cycle. (C) 2004 Elsevier Inc. All rights reserved.

    DOI

  • Effect of lithium on the circadian rhythms of locomotor activity and glycogen synthase kinase-3 protein expression in the mouse suprachiasmatic nuclei

    E Iwahana, M Akiyama, K Miyakawa, A Uchida, J Kasahara, K Fukunaga, T Hamada, S Shibata

    EUROPEAN JOURNAL OF NEUROSCIENCE   19 ( 8 ) 2281 - 2287  2004.04

     View Summary

    A circadian clock located in the suprachiasmatic nucleus (SCN) regulates the period of physiological and behavioural rhythms to approximately 24 h. Lithium can lengthen the period of circadian rhythms in most organisms although little is known about the underlying mechanism. In the present study, we examined Drosophila shaggy ortholog glycogen synthase kinase-3 (GSK-3) protein expression in the SCN after lithium treatment. When locomotor activity was assessed, we found an association between the effect of lithium and the period of circadian oscillation as well as the level of GSK-3 protein expression. The decreased expression of GSK-3 and increased expression of phosphorylated GSK-3 (pGSK-3) resulted in an antiphasic circadian rhythm between the two in the SCN of lithium-treated mice housed under both light-dark and constant dark conditions. The enzyme activity of GSK-3 in the SCN was low when the level of pGSK-3 protein was high, as examined by immunoblotting analysis. Thus, GSK-3 enzyme activity has a correlation with the expression of GSK-3 protein in the SCN. Although both GSK-3 and pGSK-3 proteins are also expressed in the arcuate nucleus, lithium did not affect their expression. Based on the association that we found between lengthened circadian period and GSK-3 protein and GSK-3 activity in the SCN, we suggest that GSK-3 plays a role in regulating the period of the mammalian circadian pacemaker.

    DOI

  • Two-peaked synchronization in day/night expression rhythms of the fibrinogen gene cluster in the mouse liver

    E Sakao, A Ishihara, K Horikawa, M Akiyama, M Arai, M Kato, N Seki, K Fukunaga, A Shimizu-Yabe, K Iwase, S Ohtsuka, T Sato, Y Kohno, S Shibata, M Takiguchi

    JOURNAL OF BIOLOGICAL CHEMISTRY   278 ( 33 ) 30450 - 30457  2003.08

     View Summary

    Genes expressed with day/night rhythms in the mouse liver were searched for by microarray analysis using an in-house array harboring mouse liver cDNAs. The rhythmic expression with a single peak and trough level was confirmed by RNA blot analysis for 3beta-Hsd and Gabarapl1 genes exhibiting a peak in the light phase and Spot14, Hspa8, Hspa5, and Hsp84-1 genes showing a peak in the dark phase. On the other hand, mRNA levels for all of the three fibrinogen subunits, Aalpha, Bbeta and gamma, exhibited two peaks each in the light and dark phases in a synchronized manner. This two-peaked rhythmic pattern of fibrinogen genes as well as the single peak-trough pattern of other genes was diminished or almost completely lost in the liver of Clock mutant mice, suggesting that the two-peaked expression is also under the control of oscillation-generating genes. In constant darkness, the first peak of the expression rhythm of fibrinogen genes was almost intact, but the second peak disappeared. Therefore, although the first peak in the subjective day is a component of the innate circadian rhythm, the second peak seems to require light stimuli. Fasting in constant darkness caused shifts of time phases of the circadian rhythms. Protein levels of the fibrinogen subunits in whole blood also exhibited circadian rhythms. In the mouse and human loci of the fibrinogen gene cluster, a number of sequence elements resembling circadian transcription factor-binding sites were found. The fibrinogen gene locus provides a unique system for the study of two-peaked day/night rhythms of gene expression in a synchronized form.

    DOI

  • セロトニンをめぐって 注目される基礎的知見 サーカディアンリズムとセロトニン

    浜田俊幸, 柴田重信

    Clin Neurosci   21 ( 6 ) 635 - 637  2003.06

    J-GLOBAL

  • Adrenergic regulation of clock gene expression in mouse liver

    H Terazono, T Mutoh, S Yamaguchi, M Kobayashi, M Akiyama, R Udo, S Ohdo, H Okamura, S Shibata

    PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA   100 ( 11 ) 6795 - 6800  2003.05

     View Summary

    A main oscillator in the suprachiasmatic nucleus (SCN) conveys circadian information to the peripheral clock systems for the regulation of fundamental physiological functions. Although polysynaptic autonomic neural pathways between the SCN and the liver were observed in rats, whether activation of the sympathetic nervous system entrains clock gene expression in the liver has yet to be understood. To assess sympathetic innervation from the SCN to liver tissue, we investigated whether injection of adrenaline/noradrenaline (epinephrine/norepinephrine) or sympathetic nerve stimulation could induce mPer gene expression in mouse liver. Acute administration of adrenaline or noradrenaline increased mPer1 but not mPer2 expression in the liver of mice in vivo and in hepatic slices in vitro. Electrical stimulation of the sympathetic nerves or adrenaline injection caused an elevation of bioluminescence in the liver area of transgenic mice carrying mPer1 promoter-luciferase. Under a light-dark cycle, destruction of the SCN flattened the daily rhythms of not only mPer1, mPer2, and mBmal1 genes but also noradrenaline content in the liver. Daily injection of adrenaline, administered at a fixed time for 6 days, recovered oscillations of mPer2 and mBmal1 gene expression in the liver of mice with SCN lesion on day 7. Sympathetic nerve denervation by 6-hydroxydopamine flattened the daily rhythm of mPer1 and mPer2 gene expression. Thus, on the basis of the present results, activation of the sympathetic nerves through noradrenaline and/or adrenaline release was a factor controlling the peripheral clock.

    DOI

  • MAP kinase-dependent induction of clock gene expression by alpha(1)-adrenergic receptor activation

    M Akiyama, Y Minami, K Kuriyama, S Shibata

    FEBS LETTERS   542 ( 1-3 ) 109 - 114  2003.05

     View Summary

    While peripheral oscillators can be reset by Immoral factors such as glucocorticoid hormones, indirect neural communications involving sympathetic and parasympathetic neurons from the suprachiasmatic nucleus to various peripheral tissues suggest that autonomic nerve innervations also function in the resetting and synchronization of peripheral tissues. To study the role of sympathetic adrenergic signaling on clock gene expression, we constructed NIH3T3 cells that stably expressed each of three alpha(1)-adrenergic receptor subtypes (alpha(1A), alpha(1B) and alpha(1D)). We found that noradrenaline transiently induced the expression of mPer1, mPer2, and mE4bp4 1-2 h after alpha(1)-receptor activation. The extent and time course of clock gene mRNA induction by noradrenaline or the alpha(1)-receptor agonist phenylephrine (PE) was similar to that seen by 50% horse serum shock. Clock gene mRNA induction by PE was inhibited by U0126, a MEK inhibitor, suggesting involvement of the mitogen-activated protein kinase signaling pathway. We also found that both mPer1 and mPet-2 mRNAs were induced in the mouse liver 60 min after PE injection. These results suggest that although humoral factors are important for entrainment of the peripheral clock, the autonomic nervous system may also be involved in the process. (C) 2003 Published by Elsevier Science B.V. on behalf of the Federation of European Biochemical Societies.

    DOI

  • Facilitation of alpha-amino-3-hydroxy-5-methylisoxazole-4-propionate receptor transmission in the suprachiasmatic nucleus by aniracetam enhances photic responses of the biological clock in rodents

    T Moriya, M Ikeda, K Teshima, R Hara, K Kuriyama, T Yoshioka, CN Allen, S Shibata

    JOURNAL OF NEUROCHEMISTRY   85 ( 4 ) 978 - 987  2003.05

     View Summary

    This study was designed to test whether the alpha-amino-3-hydroxy-5-methylisoxazole-4-propionate (AMPA) receptor-facilitating drug, aniracetam, could potentiate photic responses of the biological clock in the suprachiasmatic nucleus (SCN) of rodents. Using the whole-cell patch technique, we first demonstrated that AMPA currents elicited by either local AMPA application or optic chiasm stimulation were augmented by aniracetam in the neurons of the SCN. The AMPA application-elicited increase of intracellular Ca2+ concentration in SCN slices was also enhanced by aniracetam treatment. The systemic injection of aniracetam dose-dependently (10-100 mg/kg) potentiated the phase delay in behavioral rhythm induced by brief light exposure of low intensity (3 lux) but not high intensity (10 or 60 lux) during early subjective night. Under the blockade of NMDA receptors by (+) MK801, aniracetam failed to potentiate a light (3 lux)-induced phase delay in behavioral rhythm. Aniracetam increased the photic induction of c-Fos protein in the SCN that was elicited by low intensity light exposure (3 lux). These results suggest that AMPA receptor-mediated responses facilitated by aniracetam can explain enhanced photic responses of the biological clock in the SCN of rodents.

    DOI

  • Melatonin modulates the light-induced sympathoexcitation and vagal suppression with participation of the suprachiasmatic nucleus in mice

    T Mutoh, S Shibata, HW Korf, H Okamura

    JOURNAL OF PHYSIOLOGY-LONDON   547 ( 1 ) 317 - 332  2003.02

     View Summary

    In mammals, the autonomic nervous system mediates the central circadian clock oscillation from the suprachiasmatic nucleus (SCN) to the peripheral organs, and controls cardiovascular, respiratory and gastrointestinal functions. The present study was conducted in mice to address whether light signals conveyed to the SCN can control peripheral autonomic functions, and further examined the impact of centrally administered melatonin on peripheral autonomic functions via activation of melatonin receptor signalling. In vivo electrophysiological techniques were performed in anaesthetised, open-chest and artificially ventilated mice whilst monitoring the arterial blood pressure and heart rate. Light induced an increase of the renal sympathetic nerve activity, arterial blood pressure and heart rate immediately after lights on. Conversely, light rapidly suppressed the gastric vagal parasympathetic nerve activity, which was affected neither by hepatic vagotomy nor by total subdiaphragmatic vagotomy. These autonomic responses were mediated by the SCN since bilateral SCN lesion totally abolished the light-evoked neuronal and cardiovascular responses. Melatonin administered intracerebroventricularly (i.c.v.) attenuated the sympathetic and vagal nerve activities in a dose-dependent manner with a threshold of 0.1 ng and these effects were blocked by i.c.v. pre-treatment of the competitive melatonin receptor antagonist luzindole. These results suggest that light induces sympathoexcitation and vagal suppression through the SCN and that melatonin modulates the light-induced autonomic responses via activation of the central melatonin receptor signalling.

    DOI

  • Temporal anticipatory behavior and circadian clock

    Toshiyuki Hamada, Shigenobu Shibata

    Brain and Nerve   55 ( 1 ) 26 - 33  2003.01

    PubMed

  • Constant light housing attenuates circadian rhythms of mPer2 mRNA and mPER2 protein expression in the suprachiasmatic nucleus of mice

    M Sudo, K Sasahara, T Moriya, M Akiyama, T Hamada, S Shibata

    NEUROSCIENCE   121 ( 2 ) 493 - 499  2003

     View Summary

    Constant light (LL) or constant dark (DD) environmental lighting conditions cause a free-running period and activity reduction in the rodent behavioral circadian rhythm. In order to understand the molecular process underlying behavioral rhythms in LL or DD housing conditions, we examined the circadian profile of mPer2 mRNA and mPER2 in the suprachiasmatic nucleus (SCN), a main oscillator, of free-running mice. The circadian expression rhythm of mPer2 in the SCN was dampened under 7-day LL conditions, whereas that of mPER2 protein was moderately attenuated and its expression peak delayed. The circadian expression of mPer2 and its product was slightly attenuated and advanced by 7-day DD conditions. With arrhythmic behavioral activity caused by long-term LL housing, mPER2 protein lost its rhythmicity in the SCN. On the other hand, ILL or DD housing did not affect the mPer2 gene and its product in the cerebral cortex. The present results suggest that mPER2 circadian expression in the SCN corresponds well with behavioral circadian oscillation under LL or DD conditions. Thus, the behavioral circadian rhythm seems to correlate with molecular clock works in the SCN. (C) 2003 IBRO. Published by Elsevier Ltd. All rights reserved.

    DOI

  • 睡眠障害と体内時計 3 時刻認知学習と体内時計

    浜田俊幸, 柴田重信

    Brain Nerve   55 ( 1 ) 26 - 33  2003.01

    J-GLOBAL

  • Ca2+/calmodulin-dependent protein kinase II-dependent long-term potentiation in the rat suprachiasmatic nucleus and its inhibition by melatonin

    K Fukunaga, K Horikawa, S Shibata, Y Takeuchi, E Miyamoto

    JOURNAL OF NEUROSCIENCE RESEARCH   70 ( 6 ) 799 - 807  2002.12

     View Summary

    We recently reported that Ca2+/calmodulin-dependent protein (CaM) kinase 11 is involved in light-induced phase delays and Per gene induction in the suprachiasmatic nucleus (SCN). To clarify the activation mechanisms of CaM kinase 11 by glutamate receptor stimulation in the SCN, we documented CaM kinase 11 activation following induction of long-term potentiation (LTP) in the rat SCN. High-frequency stimulation (100 Hz, 1 sec) applied to the optic nerve resulted in LTP of a postsynaptic field potential in the rat SCN. Unlike LTP in the hippocampal CA1 region, LTP onset in the SCN was slow and partly dependent on N-methyl-D-aspartate receptor activation. LTP induction in the SCN was completely inhibited by treatment with a nitric oxide synthase inhibitor or with a specific CaM kinase 11 inhibitor. Immunoblotting analysis using phosphospecific antibodies against autophosphorylated CaM kinase 11 revealed that LTP induction was accompanied by an increase in autophosphorylation. After high-frequency stimulation, we could visualize activation of CaM kinase 11 in vasoactive intestinal polypeptide-positive neurons in the SCN by immunohistochemistry. Treatment with cyclosporin A, a calcineurin inhibitor, potentiated LTP induction in the rat SCN. Interestingly, treatment with melatonin totally prevented LTP induction, without changes in basal synaptic transmission. Analyses of phosphorylation of CaM kinase 11, mitogen-activated protein kinase, and cAMP-responsive element binding protein revealed that stimulatory and inhibitory effects on CaM kinase 11 autophosphorylation underlie the effects of cyclosporin A and melatonin, respectively. These results suggest that CaM kinase 11 plays critical roles in LTP induction in the SCN and that melatonin has inhibitory effects on synaptic plasticity through CaM kinase 11. (C) 2002 Wiley-Liss, Inc.

    DOI

  • Time-dependent effect of glutamate on long-term potentiation in the suprachiasmatic nucleus of rats

    Y Nisikawa, T Shimazoe, S Shibata, S Watanabe

    JAPANESE JOURNAL OF PHARMACOLOGY   90 ( 2 ) 201 - 204  2002.10

     View Summary

    The effect of glutamate on optic nerve stimulation-evoked field potentials in rat suprachiasmatic nucleus (SCN) was examined in vitro. Glutamate application for 20 min induced long-term potentiation (LTP) of the field potentials in the SCN at nighttime, whereas that induced a weak LTP at daytime. On the other hand, application for 40 min induced LP in the SCN during the daytime, whereas it induced a weak one at nighttime. These results indicate that the effect of glutamate is dependent on the application time and that the effect is influenced by the duration of glutamate exposure.

    DOI

  • Methamphetamine-induced, suprachiasmatic nucleus-independent circadian rhythms of activity and mPer gene expression in the striatum of the mouse

    M Iijima, T Nikaido, M Akiyama, T Moriya, S Shibata

    EUROPEAN JOURNAL OF NEUROSCIENCE   16 ( 5 ) 921 - 929  2002.09

     View Summary

    While the suprachiasmatic nucleus (SCN) coordinates the majority of daily rhythms, some circadian patterns of expression are controlled from outside of the SCN. These include responses to daily methamphetamine (MAP) injection, or daily restricted feeding. The mechanisms underlying these SCN-independent circadian rhythms are unknown. A circadian oscillation in the expression of mPer1 and/or mPer2 , mouse period, in the SCN is considered necessary to generate an SCN-dependent circadian rhythm. Therefore, in this experiment, we examined the association between mPer gene expression and the MAP-induced, SCN-independent circadian rhythm. Acute injection of MAP caused an elevation of mPer1, mBmal1, and mNpas2 gene expression in the striatum and mPer1 in the liver. Daily MAP injection at a fixed time for 6 days shifted the rhythmic mPer1 and mPer2 expression in the striatum from a nocturnal to a diurnal rhythm, but failed to affect that in the SCN. Although lesion of the SCN 'flattened'mPer gene oscillation in the striatum and liver, daily MAP injection caused both behavioural and mPer gene expression rhythms. Daily MAP injection at variable injection intervals (12-36 h) for 6 days, however, failed to produce mPer gene rhythm in the striatum. Daily repeated MAP signals may strengthen the oscillatory force of SCN-independent circadian behavioural and molecular rhythms. The present results suggest that daily oscillation of mPer genes outside the SCN is closely associated with the regulation of SCN-independent rhythms. Thus, the present experiment highlights strongly the important role of clock gene expression, in the brain, that underlies the circadian behavioural rhythm.

    DOI

  • Restricted feeding induces daily expression of clock genes and Pai-1 mRNA in the heart of Clock mutant mice

    Y Minami, K Horikawa, M Akiyama, S Shibata

    FEBS LETTERS   526 ( 1-3 ) 115 - 118  2002.08

     View Summary

    Plasminogen activator inhibitor-1 (PAI-1) is a key factor of fibrinolytic activity. The activity and mRNA abundance show a daily rhythm. To elucidate the mechanism of daily Pai-1 gene expression, the expression of Pai-1 and several clock genes was examined in the heart of homozygous Clock mutant (Clock/Clock) mice. Damping of the daily oscillation of Pai-1 gene expression in Clock/Clock mice was accompanied with damped or attenuated oscillations of mPer1, mPer2, mBmal1, and mNpas2 mRNA. Daily restricted feeding induced a daily mRNA rhythm of all clock genes and Pai-1 mRNA in Clock/Clock mice as well as wild-type mice. The peaks of clock genes and Pai-1 mRNA were phase-advanced in the heart of both genotypes after 6 days of restricted feeding. The present results demonstrate that daily Pai-1 gene expression depends on clock gene expression in the heart and that a functional Clock gene is not required for restricted feeding-induced resetting of the peripheral clock. (C) 2002 Federation of European Biochemical Societies. Published by Elsevier Science B.V. All rights reserved.

    DOI

  • Extended action of MKC-242, a selective 5-HT1A receptor agonist, on light-induced Per gene expression in the suprachiasmatic nucleus in mice

    S Takahashi, Y Yoshinobu, R Aida, H Shimomura, M Akiyama, T Moriya, S Shibata

    JOURNAL OF NEUROSCIENCE RESEARCH   68 ( 4 ) 470 - 478  2002.05

     View Summary

    We reported previously that (S)-5-[3-[(1,4-benzodioxan-2ylmethyl)amino]propoxy]-1,3-benzodioxole hydrochloride (MKC-242) (3 mg kg(-1), i.p.), a selective 5-HT1A receptor agonist, accelerated the re-entrainment of hamster wheel-running rhythms to a new 8 hr delayed or advanced light-dark cycle, and also potentiated the phase advance of the wheel-running rhythm produced by light pulses. The molecular mechanism underlying MKC-242-induced potentiation of this phase shift, however, has not yet been elucidated. We examined the effects of MKC-242 on light-induced mPer1 and mPer2 mRNA expression in the suprachiasmatic nucleus (SCN) of mice. MKC-242 (5 mg kg(-1), i.p.) potentiated light-induced mPer1 and mPer2 expression in the SCN of mice housed in constant darkness for 2 days, when mRNA levels were observed 3 hr after light-exposure. More potentiating action of MKC-242 on mPet2 expression in the SCN was observed in mice housed in constant darkness for 9-10 days. This facilitatory action of MKC-242 on mPer1 expression was antagonized by WAY100635, a selective 5-HT1A receptor blocker, indicating that MKC-242 activated 5-HT1A receptors. Other drugs such as 8-hydroxy-dipropylaminotetralin (10 mg kg(-1),p.), paroxetine (10 mg kg(-1), i.p.), buspirone (10 mg kg(-1) i.p.), and diazepam (10 mg kg(-1), i.p.) did not display a potentiating action on light-induced mPer1 and mPer2 expression in the SCN. In the behavioral experiments, we found that MKC-242 (5 mg kg(-1), i.p.) potentiated lightinduced phase delays of free-running rhythm in mice. The present results suggest that prolonged increase of mPer1 or mPer2 expression in the SCN by MKC-242 may be involved in the potentiation of photic entrainment by MKC242 in mice. (C) 2002 wiley-Liss, Inc.

    DOI

  • Motor discoordination in mutant mice lacking junctophilin type 3

    M Nishi, K Hashimoto, K Kuriyama, S Komazaki, M Kano, S Shibata, H Takeshima

    BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS   292 ( 2 ) 318 - 324  2002.03

     View Summary

    Junctional complexes between the plasma membrane and endoplasmic reticulum (ER), often called "subsurface cisternae" or "peripheral coupling," are shared by excitable cells. These junctional membranes probably provide structural foundation for functional crosstalk between cell-surface and intracellular ionic channels. Our current studies have indicated that junctophilins (JPs) take part in the formation of functional membrane complexes by spanning the ER membrane and interacting with the plasma membrane. Of the JP subtypes defined, JP type 3 (JP-3) is specifically expressed in neurons in the brain. It has been currently reported that triplet repeat expansions in the JP-3 gene are associated with Huntington's disease-like symptoms including motor disorder in human. To survey the physiological role of JP-3, we generated the knockout mice. The JP-3-knockout mice grew and reproduced normally, and we did not observe any morphological abnormality in the mutant brain. In the behavioral study, the mutant mice showed impaired performance specifically in balance/motor coordination tasks. Although obvious defects could not be observed in excitatory transmission among cerebellar neurons from the mutant mice, the data indicate that JP-3 plays an active role in certain neurons involved in motor coordination. (C) 2002 Elsevier Science (USA).

    DOI

  • 体内時計と疾患

    島田和幸, 柴田重信, 前村浩二

    現代医療   34 ( 6 )  2002

  • Pituitary adenylate cyclase-activating polypeptide produces a phase shift associated with induction of mPer expression in the mouse suprachiasmatic nucleus

    Y Minami, K Furuno, M Akiyama, T Moriya, S Shibata

    NEUROSCIENCE   113 ( 1 ) 37 - 45  2002

     View Summary

    The main mammalian circadian pacemaker is located in the suprachiasmatic, nucleus of the hypothalamus. Clock genes such as the mouse Period gene (mPer) play a role in this core clock mechanism in the mouse. With brief light exposure during the subjective night, the photic information, which is conveyed directly to the suprachiasmatic nucleus via the retinohypothalamic tract, results in mPer1 and mPer2 expression in the suprachiasmatic nucleus.
    Glutamate and pituitary adenylate cyclase-activating polypeptide (PACAP) are co-stored in the retinohypothalamic tract. Recent studies have suggested that not only glutamate but also PACAP are key players in the phase shift that occurs during subject night; however, research demonstrating a direct association between the PACAP-induced phase shift and mPer gene expression has yet to be conducted.
    In the present study, PACAP (200 pmol) injected into the lateral ventricle during subjective night (circadian time 16; circadian time 12, onset of locomotor activity) caused a moderate phase delay associated with moderate expression of mPer1 and only slight expression of mPer2 in the mouse suprachiasmatic nucleus. PACAP-induced mPer1 expression was also observed in the paraventricular nucleus and periventricular area of the hypothalamus. (+)MK-801 (0.5 mg/kg), an N-methyl-D-aspartate (NMDA) receptor antagonist, suppressed both the PACAP-induced phase delay and mPer1 expression. From these results we suggest that PACAP induces phase delays in the mouse circadian rhythm in association with ;an increase of mPer expression in the suprachiasmatic nucleus via the activation of NMDA receptors. (C) 2002 IBRO. Published by Elsevier Science Ltd. All rights reserved.

    DOI

  • Gastrin-releasing peptide mediates photic entrainable signals to dorsal subsets of suprachiasmatic nucleus via induction of Period gene in mice

    R Aida, T Moriya, M Araki, M Akiyama, K Wada, E Wada, S Shibata

    MOLECULAR PHARMACOLOGY   61 ( 1 ) 26 - 34  2002.01

     View Summary

    The suprachiasmatic nucleus (SCN), locus of the central circadian clock, consists of two neuronal populations (i.e., a light-recipient ventral SCN subpopulation directly entrained by light and a dorsal SCN subpopulation with an autonomous oscillatory function possessing an indirect or weak light response). However, the mechanism underlying the transmission of photic signals from the ventral to dorsal SCN remains unclear. Because gastrin-releasing peptide (GRP), expressed mainly in the ventral SCN, exerts phase-shifting actions, loss of the GRP receptor intuitively implies a reduction of photic information from the ventral to dorsal SCN. Therefore, using GRP receptor-deficient mice, we examined the involvement of GRP and the GRP receptor in light- and GRP-induced entrainment by the assessment of behavioral rhythm and induction of mouse-Period (mPer) gene in the SCN, which is believed to be a critical for photic entrainment. Administration of GRP during nighttime dose dependently produced a phase delay of behavior in wildtype but not GRP receptor-deficient mice. This phase-shift by GRP was closely associated with induction of mPer1 and mPer2 mRNA as well as c-Fos protein in the dorsal portion of the SCN, where the GRP receptor was also expressed abundantly. Both the light-induced phase shift in behavior and the induction of mPer mRNA and c-Fos protein in the dorsal SCN were attenuated in GRP receptor-deficient mice. Our present studies suggest that GRP neurons in the retinorecipient ventral area of the SCN convey the photic entrainable signals from the ventral SCN to the dorsal SCN via induction of the mPer gene.

    DOI

  • Circadian profile of Per gene mRNA expression in the suprachiasmatic nucleus, paraventricular nucleus, and pineal body of aged rats

    M Asai, Y Yoshinobu, S Kaneko, A Mori, T Nikaido, T Moriya, M Akiyama, S Shibata

    JOURNAL OF NEUROSCIENCE RESEARCH   66 ( 6 ) 1133 - 1139  2001.12

     View Summary

    Aging alters circadian components such as the free-running period, the day-to-night activity ratio and photic entrainment in behavioral rhythms, and 2-deoxyglucose uptakes and neuronal firing in the suprachiasmatic nucleus (SCN). A core clock mechanism in the mouse SCN appears to involve a transcriptional feedback loop in which Period (Per) and Cryptochrome (Cry) genes play a role in negative feedback. The circadian rhythm systems include photic entrainment, clock oscillation, and outputs of clock information such as melatonin production. In this experiment, we examined clock gene expression to determine whether circadian input, oscillation, and output are disrupted with aging. Circadian expression profiles of rPer1, rPer2, or rCry1 mRNA were very similar in the SCN, the paraventricular nucleus of the hypothalamus (PVN), and the pineal body of young and aged (22-26 months) rats. On the other hand, the photic stimulation-induced rapid expression of Per1 and Per2 in the SCN was reduced with aging. The present results suggest that the molecular mechanism of clock oscillation in the SCN, PVN, and pineal body is preserved against aging, whereas the impairment of Per1 induction in the SCN after light stimulation may result in impaired behavioral photic entrainment in aged rats. (C) 2001 Wiley-Liss, Inc.

    DOI

  • 時計遺伝子に対するベンゾジアゼピン受容体作用薬の影響

    柴田重信

    老年精神医学雑誌   12 ( 11 ) 1344 - 1350  2001.11

  • Physical and inflammatory stressors elevate circadian clock gene mPer1 mRNA levels in the paraventricular nucleus of the mouse

    S Takahashi, SI Yokota, R Hara, T Kobayashi, M Akiyama, T Moriya, S Shibata

    ENDOCRINOLOGY   142 ( 11 ) 4910 - 4917  2001.11

     View Summary

    Stress induces secretion of corticosterone through activation of hypothalamic-pituitary-adrenal axis. This corticosterone the hy secretion is thought to be controlled by a circadian clock in the suprachiasmatic nucleus (SCN). The hypothalamic paraventricular nucleus (PVN) receives convergent information from both stress and the circadian clock. Recent reports demonstrate that mammalian orthologs (Per1, Per2, and Per3) of the Drosophila clock gene Period are expressed in the SCN, PVN, and peripheral tissues. In this experiment, we examined the effect of physical and inflammatory stressors on mPer gene expression in the SCN, PVN, and liver. Forced swimming, immobilization, and lipopolysaccharide injection elevated mPer1 gene expression in the PVN but not in the SCN or liver. A stress-induced increase in mPer1 expression was observed in the corticotropin-releasing factor-positive cells of the PVN; however, the stressors used in this study did not affect mPer2 expression in the PVN, SCN, or liver. The present study suggests that a stress-induced disturbance of circadian corticosterone secretion may be associated with the stress-induced expression of mPer1 mRNA in the PVN.

    DOI

  • Visualization of mPer1 transcription in vitro: NMDA induces a rapid phase shift of mPer1 gene in cultured SCN

    M Asai, S Yamaguchi, H Isejima, M Jonouchi, T Moriya, S Shibata, M Kobayashi, H Okamura

    CURRENT BIOLOGY   11 ( 19 ) 1524 - 1527  2001.10

     View Summary

    Many physiological and behavioral phenomena are controlled by an internal, self-sustaining oscillator with a periodicity of approximately 24 hr. In mammals, the principal oscillator resides in the suprachiasmatic nucleus (SCN). A light pulse during the subjective night causes a phase shift of the circadian rhythm via direct glutamatergic retinal afferents to the SCN [1]. Along with the accepted theoretical models of the clock, it is suggested that behavioral resetting of mammals is completed within 2 hr [2]; however, the molecular mechanism has not been elucidated. Here, we show the realtime image of the transcription of the circadian-clock gene mPer1 in the cultured SCN by using the transgenic. mice that carry a luciferase reporter gene under the control of the mPer1 promoter [3]. The real-time image demonstrates that the mPer1 promoter activity oscillates robustly in a circadian manner and that this promoter activity is reset rapidly (within 2-3 hr) when a phase shift occurs.

    DOI

  • Calcium and pituitary adenylate cyclase-activating polypeptide induced expression of circadian clock gene mPer1 in the mouse cerebellar granule cell culture

    M Akiyama, Y Minami, T Nakajima, T Moriya, S Shibata

    JOURNAL OF NEUROCHEMISTRY   78 ( 3 ) 499 - 508  2001.08

     View Summary

    Mammalian circadian clock genes Per1 and Per2 are rhythmically expressed not only in the suprachiasmatic nucleus where the mammalian circadian clock exists, but also in other brain regions and peripheral tissues. The induced circadian oscillation of Per genes after treatment with high concentrations of serum or various drugs in cultured cells suggests the ubiquitous existence of the oscillatory mechanism. These treatments also result in a rapid surge of expression of Per1. It has been shown that multiple signaling pathways are involved in Per1 gene induction in culture cells. We used a dispersed primary cell culture made up of mouse cerebellar granule cells to examine the stimuli inducing the mPer genes and their signaling pathways in neuronal tissues expressing mPer genes. We demonstrated that mPer1, but not mPer2, mRNA expression was dependent on the depolarization state controlled by extracellular KCI concentration in the granule cell culture. Nifedipine treatment reduced mPer1 induction, suggesting that mPer1 mRNA expression depends on intracellular calcium concentration regulated through a voltage-dependent Ca2+ channel. Transient mPer1 mRNA induction was observed after elevating KCI concentration in the medium from 5 mm to 25 mm. This increased expression was suppressed by a calmodulin antagonist, or CaMKII/IV inhibitor, but not by MEK inhibitors. Addition of pituitary adenylate cyclase-activating polypeptide-38 to the medium also induced transient Per1 gene expression. This induction was mimicked by dibutyryl-cAMP and suppressed by a protein kinase A (PKA) inhibitor, but not by MEK inhibitors. These results suggest that Ca2+/calmodulin-dependent protein kinase II/IV- and PKA-dependent pathways are involved in high-KCI and PACAP-induced mPer1 induction, respectively, and neural tissues use multiple signaling pathways for mPer1 induction similar to culture cells.

    DOI

  • 視交叉上核における時計リセッティングのシグナル伝達

    守屋孝洋, 柴田重信

    細胞工学   20 ( 6 ) 828 - 836  2001.06

  • Involvement of calcium-calmodulin protein kinase but not mitogen-activated protein kinase in light-induced phase delays and Per gene expression in the suprachiasmatic nucleus of the hamster

    S Yokota, M Yamamoto, T Moriya, M Akiyama, K Fukunaga, E Miyamoto, S Shibata

    JOURNAL OF NEUROCHEMISTRY   77 ( 2 ) 618 - 627  2001.04

     View Summary

    It is known that Ca2+-dependent phosphorylation of cAMP response element binding protein (CREB) and the rapid induction of mPer1 and mPer2, mouse period genes in the suprachiasmatic nucleus (SCN) are associated with light-induced phase shifting. The CREB/CRE transcriptional pathway has been shown to be activated by calcium/calmodulin dependent kinase II (CaMKII) and mitogen-activated protein kinase (MAPK); however, there is a lack of evidence concerning whether the activation of CaMKII and/or MAPK elicited by photic stimuli are associated with the change in Per gene expression and behavioral phase shifting. In this experiment, we found there was an inhibitory effect by KN93, CaMKII inhibitor, on hamster Per1 and Per2 expression in the SCN and on phase delays in wheel running rhythm induced by light pulses. PD98059 and U0126, MAPK kinase inhibitors, however, affected neither light-induced Per1 and Per2 expression nor behavioral phase delays, even though PD98059 attenuated the light-induced phosphorylation of MAPK in the SCN. The present findings demonstrate that the light-induced activation of CaMKII plays an important role in the induction of Per1 and Per2 mRNA in the hamster SCN as well as phase shifting. These results suggest that gated induction of Per1 and/or Per2 genes through CaMKII-CREB/CRE accompanied with photic stimuli may be a critical step in phase shifting.

    DOI

  • Sensitized increase of period gene expression in the mouse caudate/putamen caused by repeated injection of methamphetamine

    T Nikaido, M Akiyama, T Moriya, S Shibata

    MOLECULAR PHARMACOLOGY   59 ( 4 ) 894 - 900  2001.04

     View Summary

    Methamphetamine (MAP) causes the sensitization phenomena not only in MAP-induced locomotor activity, dopamine release, and Fos expression, but also in MAP-induced circadian rhythm. Cocaine-induced sensitization is reportedly impaired in Drosophila melanogaster mutant for the Period (Per) gene. Thus, sensitization may be related to induction of the Per gene. A rapid induction of mPer1 and/or mPer2 in the suprachiasmatic nucleus after light exposure is believed to be necessary for light-induced behavioral phase shifting. Although the caudate/putamen (CPu) expresses mPer1 and/or mPer2 mRNA, the function of these genes in this nucleus has not yet been elucidated. Therefore, we examined whether MAP affects the expression of mPer1 and/or mPer2 mRNA in the mouse CPu. Injection of MAP augmented the expression of mPer1 but not mPer2 or mPer3 in the CPu, and this MAP-induced increase in mPer1 expression lasted for 2 h. Also, the MAP-induced increase of mPer1 mRNA was strongly antagonized by pretreatment with a dopamine D1 receptor and N-methyl-D-aspartate (NMDA) receptor antagonist, but not by a D2 receptor antagonist. Interestingly, application of either the D1 or the D2 agonist alone did not cause mPer1 expression. The present results demonstrate that activation of both NMDA and D1 receptors is necessary to produce MAP-induced mPer1 expression in the CPu. Repeated injection of MAP caused a sensitization in not only the locomotor activity but also mPer1 expression in the CPu without affecting the level of mPer2, mPer3, or mTim mRNA. Thus, these results suggest that MAP-induced mPer1 gene expression may be related to the mechanism for MAP-induced sensitization in the mouse.

  • Restricted feeding entrains liver clock without participation of the suprachiasmatic nucleus

    R Hara, KK Wan, H Wakamatsu, R Aida, T Moriya, M Akiyama, S Shibata

    GENES TO CELLS   6 ( 3 ) 269 - 278  2001.03

     View Summary

    Background: There are two main stimuli that entrain the circadian rhythm, the light-dark cycle (LD) and restricted feeding (RF). Light-induced entrainment requires induction of the Per1 and Per2 genes in the suprachiasmatic nucleus (SCN), the locus of a main oscillator. In this experiment, we determined whether RF resets the expression of circadian clock genes in the mouse liver with or without participation of the SCN.
    Results: Mice were allowed access to food for 4 h during the daytime (7 h advance of feeding time) under LD or constant darkness (DD). The peaks of mPer1, mPer2, D-site-binding protein (Dbp) and cholesterol 7 alpha -hydroxylase (Cyp7A) mRNA in the liver were advanced 6-12 h after 6 days of RF, whereas those in SCN were unaffected. The advance of mPer expression in the liver by RF was still observed in SCN-lesioned mice. A 7 h advance in the LD cycle advanced the peaks of clock gene expression in both the liver and SCN, whereas, a shift in the LD did not move the phase of the liver clock when the shift was carried out under a fixed RF schedule during the night-time.
    Conclusions: These results suggest that restricted feeding strongly entrained the expression of circadian clock genes in the liver without the participation of an SCN clock function.

    DOI

  • Restricted-feeding-induced anticipatory activity rhythm is associated with a phase-shift of the expression of mPer1 and mPer2 mRNA in the cerebral cortex and hippocampus but not in the suprachiasmatic nucleus of mice

    H Wakamatsu, Y Yoshinobu, R Aida, T Moriya, M Akiyama, S Shibata

    EUROPEAN JOURNAL OF NEUROSCIENCE   13 ( 6 ) 1190 - 1196  2001.03

     View Summary

    Daily restricted feeding (RF) can produce food-entrainable oscillations in both intact and suprachiasmatic nucleus (SCN)-lesioned animals. Thus, there are two circadian rhythms, one of which is SCN-dependent and the other SCN-independent. Recently, it has been established that several mouse clock genes, such as mPer1, mPer2 and mPer3 are expressed in the SCN and other brain tissues. Although the role of mPer genes expressed in the SCN has recently been evaluated in the SCN-dependent rhythm, their function in the SCN-independent rhythm is still poorly understood. In order to understand the role of these genes in SCN-independent rhythm, we examined the expression pattern of mPer1 and mPer2 mRNA in each brain area of mice under RF. Mice were allowed access to food for 4 h during either the daytime under a light-dark cycle or the subjective daytime under constant dark. After 6 days of scheduled RF, the night-time or subjective night-time peak of mPer mRNA changed to a daytime peak in the cerebral cortex and hippocampus, with moderate expression in the striatum, pyriform cortex and paraventricular nucleus, and no expression in the SCN. The daytime peak in the cerebral cortex returned to a night-time peak after the release of RF to a free-feeding schedule. Although the basal rhythm of mPer expression disappeared in SCN-lesioned mice, RF produced mPer mRNA rhythm in the cerebral cortex of these mice. The present results provide evidence of an association between food-entrainable oscillations and the expression of mPer1 and mPer2 in the cerebral cortex and hippocampus.

    DOI

  • Expression of the Per1 gene in the hamster: Brain atlas and circadian characteristics in the suprachiasmatic nucleus

    S Yamamoto, Y Shigeyoshi, Y Ishida, T Fukuyama, S Yamaguchi, K Yagita, T Moriya, S Shibata, N Takashima, H Okamura

    JOURNAL OF COMPARATIVE NEUROLOGY   430 ( 4 ) 518 - 532  2001.02

     View Summary

    Recent progress in study an the molecular component of mammalian clocks has claimed that mammals and Drosophila share the similar fundamental clock oscillating system. In the present study, we investigated expression of Per1, the first gene of the mammalian homolog of the Drosophila clock gene period, in the hamster brain, and we also examined its circadian expression pattern in the mammalian clock center, the suprachiasmatic nucleus (SCN). In situ hybridization using isotope-labeled cRNA probes revealed a wide and region-specific distribution of Per1 in the hamster brain and spinal cord. High levels of Per1 were found in the internal granular layer of the granular cells of the olfactory bulb, anterior olfactory nuclei, tenia tecta, olfactory tubercle, piriform cortex,suprachiasmatic nucleus, and gyrus dentatus of hippocampus. Moderate levels of expression were detected in many brain regions including the granular layer of the cerebellum, anterior paraventricular thalamic nucleus, caudate-putamen, inferior colliculus, pontine nuclei, inferior olive, and nucleus of the solitary tract. We examined the circadian profile of hamster Per1 mRNA in the SCN in constant darkness and found that Per1 expression showed a peak at subjective day (circadian time [CT] 4) and formed a trough at subjective night (CT16-CT20). A brief exposure of light at CT16 could acutely induce large quantities of Per1 mRNA in the hamster SCN, except for its dorsomedial subdivision. These findings suggest that the characteristics of Per1 gene expression in the mammalian circadian center (showing a peak in the daytime and a trough in the nighttime and a rapid inducibility by light) are common among mammalian species. Lastly, in hamster brain, Per1 gene is also inducible in extra-SCN brain nuclei, since light at night also elicited Per1 mRNA in neurons of the hypothalamic paraventricular nucleus. (C) 2001 Wiley-Liss, Inc.

    DOI

  • Differential daily expression of Per1 and Per2 mRNA in the suprachiasmatic nucleus of fetal and early postnatal mice

    H Shimomura, T Moriya, M Sudo, H Wakamatsu, M Akiyama, Y Miyake, S Shibata

    EUROPEAN JOURNAL OF NEUROSCIENCE   13 ( 4 ) 687 - 693  2001.02

     View Summary

    It is well known that there are circadian rhythms of 2-deoxyglucose uptake and neuronal firing in the rat suprachiasmatic nucleus (SCN) during fetal and early postnatal periods. A core clock mechanism in the mouse SCN appears to involve a transcriptional feedback loop in which CLOCK and BMAL1 function as positive regulators and three mPeriod (mPer) genes play a role in negative feedback. Per genes expression occurs not only in the adult SCN but also in the fetal SCN. However, the developmental change in these genes remains unclear. In this experiment, we examined the day-night pattern of expression of Per1 and Per2 mRNA in the mouse SCN and cerebral cortex on embryonic day 17, postnatal day 3, and in young adult mice under a light-dark cycle. Daily rhythms of mRNA content were observed in mPer1 but not mPer2 in the fetal SCN. Interestingly, the expression of mPer2 in the SCN was high throughout the entire day, and a significant daily rhythm of this gene was observed on postnatal day 6. The expression pattern of SCN mPer1 in constant darkness was similar to that seen in the light-dark cycle. The present results suggest that the daily oscillation of mPer1 but not of mPer2 in the SCN in fetal and early postnatal mice may be associated with the daily rhythms of 2-deoxyglucose uptake and neuronal firing.

    DOI

  • Gene expression - View of a mouse clock gene ticking

    S Yamaguchi, M Kobayashi, S Mitsui, Y Ishida, GTJ van der Horst, M Suzuki, S Shibatall, H Okamura

    NATURE   409 ( 6821 ) 684 - 684  2001.02

    DOI

  • 概日リズムの薬理

    柴田重信

    神経研究の進歩   45   763 - 774  2001

    DOI

  • 視交叉上核と体内時計の同調機構

    秋山正志, 柴田重信

    臨床神経科学   19   55 - 57  2001

  • 光受容分子

    秋山正志, 柴田重信

    臨床検査   45   640 - 643  2001

    DOI

  • ハムスター視交叉上核の時計遺伝子(per1およびper2)発現に対するブロチゾラムの抑制作用

    柴田重信, 横田伸一

    新薬と臨床   50 ( 5 ) 15 - 25  2001

  • 東向き海外旅行における睡眠感悪化に対する出発前食事時間前進による軽減効果

    駒田陽子, 山崎勝男, 金本礼次郎, 小林強, 柴田重信

    新薬と臨床   50 ( 8 ) 780 - 788  2001

  • Additive effect of mPer1 and mPer2 antisense oligonucleotides on light-induced phase shift

    H Wakamatsu, S Takahashi, T Moriya, ST Inouye, H Okamura, M Akiyama, S Shibata

    NEUROREPORT   12 ( 1 ) 127 - 131  2001.01

     View Summary

    It is well known that light induces both mPer1 and mPer2 mRNA in the suprachiasmatic nucleus. We have reported that mPer1 antisense oligonucleotides (ODNs) inhibited the light-induced phase delays of mouse locomotor rhythm. In this study, we asked whether both or either mPer1 or mPer2 expression is necessary to induce the phase shift. We examined the effects of inhibition of mRNA expression on light-induced phase delays of mouse circadian behavior rhythm. Light-induced phase delays were moderately attenuated by microinjection of mPer1 or mPer2 antisense ODN, but not by mPer3 antisense or mPer1, mPer2 scrambled ODNs, whereas following simultaneous injection of both mPer1 and mPer2 antisense ODNs they disappeared. The present results suggest that acute induction of mPer1 and mPer2 gene play an additive effect on photic entrainment. NeuroReport 12:127-131 (C) 2001 Lippincott Williams & Wilkins.

  • Inhibitory action of brotizolam on circadian and light-induced Per1 and Per1 expression in the hamster suprachiasmatic nucleus

    S Yokota, K Horikawa, M Akiyama, T Moriya, S Ebihara, G Komura, T Ohta, S Shibata

    BRITISH JOURNAL OF PHARMACOLOGY   131 ( 8 ) 1739 - 1747  2000.12

     View Summary

    1 Triazolam reportedly causes phase advances in hamster wheel-running rhythm after injection during subjective daytime. However, it is unclear whether benzodiazepine affects the Per gene expression accompanying a behavioural phase shift.
    2 Brotizolam (0.5-10 mg kg(-1)) induced large phase advances in hamster rhythm when injected during mid-subjective daytime (circadian time 6 or 9), but not at circadian time 0, 3 or 15.
    3 Brotizolam (5 mg kg(-1)) significantly reduced the expression of Per1 and Per2 in the suprachiasmatic nucleus 1 and 2 h after injection at circadian time 6, and slightly reduced them at circadian time 20.
    4 Injection of 8-OH-DPAT (5 mg kg(-1)) at subjective daytime induced similar phase advances with a reduction of Per1 and Per2 expression. Co-administration of brotizolam with 8-OH DPAT failed to potentiate the 8-OH DPAT-induced phase advances and reduced Per expression.
    5 Both phase advance and rapid induction of Per1 and Per2 in the suprachiasmatic nucleus after light exposure (5 lux, 15 min) at circadian time 20 was strongly attenuated by co-treatment with brotizolam 5 mg kg(-1).
    6 The present results strongly suggest that reduction of Per1 and/or Per2 expression during subjective daytime by brotizolam may be an important step in causing a behavioural phase advance. The co-administration experiment suggests that common mechanism(s) are involved in brotizolamor 8-OH DPAT-induced phase advances and the reduction of Per gene expression.
    7 These results suggest that brotizolam is not only a good drug for insomnia but also a drug capable of facilitating re-entrainment like melatonin.

    DOI

  • Correlative association between N-methyl-D-aspartate receptor-mediated expression of period genes in the suprachiasmatic nucleus and phase shifts in behavior with photic entrainment of clock in hamsters

    T Moriya, K Horikawa, M Akiyama, S Shibata

    MOLECULAR PHARMACOLOGY   58 ( 6 ) 1554 - 1562  2000.12

     View Summary

    Because the rapid induction of Period (Per) genes is associated with the photic entrainment of the biological clock, we examined whether N-methyl-D-aspartate (NMDA) receptors were involved in the photic induction of Per genes in the hamster suprachiasmatic nucleus (SCN). In situ hybridization observation revealed that light during the early subjective night [circadian time (CT) 13.5] or the late subjective night (CT20) caused an induction of Per1 and Per2 but not Per3 mRNA in the SCN. Photic induction of Per mRNA at CT13.5 was observed especially in the ventrolateral SCN, whereas that at CT20 was more widespread from the ventrolateral to the dorsal SCN. A noncompetitive NMDA receptor antagonist, +MK801, dose-dependently (0.1-5.0 mg/kg) suppressed only the ventrolateral part of Per1 and Per2 mRNA induction by light at CT13.5 or CT20 in the SCN. The suppressive effects of +MK801 on Per mRNA strongly correlated with the attenuating action of this compound on phase shifts by light at both CT13.5 and CT20. A competitive NMDA receptor antagonist, D-2-amino-5-phosphonovalerate (D-APV), also exhibited inhibitory actions on light (CT20)-induced Per1 and Per2 mRNA expression in the ventrolateral SCN. Furthermore, local injection of NMDA into the SCN resulted in the induction of Per1 and Per2 mRNA in the SCN. Among NMDA receptors, NR2B and NR2C mRNA were expressed in the ventrolateral and dorsal SCN, respectively. These results suggest that the activation of NMDA receptor is a critical step for photic induction of Per1 and Per2 transcripts in the SCN, which are linked to a photic behavioral entrainment.

  • Administration of melatonin in drinking water promotes the phase advance of light-dark cycle in senescence-accelerated mice, SAMR1 but not SAMP8

    M Asai, M Ikeda, M Akiyama, Oshima, I, S Shibata

    BRAIN RESEARCH   876 ( 1-2 ) 220 - 224  2000.09

     View Summary

    We analyzed effects of aging on behavioral rhythms in the mouse showing senescence acceleration, SAMP8 strains. The free-running rhythms had longer free-running periods (tau) in SAMP8 than in the control strain (SAMR1). Drinking of melatonin promoted the adaptation to advanced LD in SAMR1 but not in SAMP8, although both strains exhibited melatonin MTI and MT2, receptors. The present results suggest that melatonin promotes the adaptation to advanced LD cycles in normal aging mice. (C) 2000 Elsevier Science B.V. All rights reserved.

    DOI

  • N-methyl-D-aspartate receptor subtype 2C is not involved in circadian oscillation or photoic entrainment of the biological clock in mice

    T Moriya, S Takahashi, M Ikeda, K Suzuki-Yamashita, M Asai, H Kadotani, H Okamura, T Yoshioka, S Shibata

    JOURNAL OF NEUROSCIENCE RESEARCH   61 ( 6 ) 663 - 673  2000.09

     View Summary

    Ishida et at. [1994: Neurosci Lett 166: 211-215] reported the circadian change of N-methyl-D-aspartate (NMDA) receptor subtype 2C mRNA and photic induction of this receptor's mRNA in the suprachiasmatic nucleus (SCN). Therefore, we investigated the role of NMDA receptor subtypes in the biological clock using NMDA receptor 2A (NR2A)- or 2C (NR2C)-deficient mice. However, NR2C-/-mice showed normal light-dark (LD)-entrained locomotor activity rhythms and free-running rhythms under constant darkness and also exhibited normal reentrainment to 6-hr LD shifts and phase delays with single light pulses. Thus, present results demonstrated no significant NR2C contribution to circadian oscillation and photic entrainment, even though expression of NR2C mRNA was highly observed in the SCN. On the other hand, the period of the free-running activity rhythm in NR2A-/mice but not NR2C-/- mice was slightly longer than that in wild-type mice in spite of low expression of NR2A in the SCN, Furthermore, reentrainment to an LD advance in NR2A-/- mice was slower under low-intensity light conditions, Thus, we suggest that NR2A plays a role in determining the behavioral state that affects the circadian rhythm. in order to elucidate the role of NR2A and NR2C in the SCN, we examined NMDA-induced Ca2+ elevations in the SCN of mutant mice using a Ca2+ imaging method, A partial reduction in Ca2+ elevation was observed in both NR2A-/- and NR2C-/- mice when high concentrations (100 or 300 mu M) of NMDA were applied. The present results suggest that NR2A plays a weak role in oscillation or entrainment of the biological clock, and that NR2C does not participate in the functions of circadian oscillation and light entrainment, (C) 2000 Wiley-Liss, Inc.

    DOI

  • Close linkage between calcium/calmodulin kinase II alpha/beta and NMDA-2A receptors in the lateral amygdala and significance for retrieval of auditory fear conditioning

    T Moriya, Y Kouzu, S Shibata, H Kadotani, K Fukunaga, E Miyamoto, T Yoshioka

    EUROPEAN JOURNAL OF NEUROSCIENCE   12 ( 9 ) 3307 - 3314  2000.09

     View Summary

    The general mechanism underlying memory and learning is an area under intense investigation and debate, yet this mechanism still remains elusive. Auditory fear conditioning (when a tone is paired with a foot shock) is a simple associative form of learning for which many mechanistic details are known. Lesions of the lateral/basolateral nuclei of the amygdala result in the selective impairment of fear conditioning, indicating that this is a key region for this type of learning. Fear conditioning induces a lasting synaptic potentiation in the lateral nuclei of the amygdala. In addition, recent results from several laboratories suggest that N-methyl-D-aspartate (NMDA) receptor activation in the amygdala is required for the acquisition and expression of cue-conditioned fear responses using several kinds of antagonists. Little is known, however, about the signal transduction pathway and molecular substrate underlying fear conditioning. Here we use NMDA receptor-deficient mice to demonstrate that calmodulin-dependent kinase II, CaMKII beta, and CaMKII alpha. activation involves the NR2A subunit in the lateral/basolateral amygdala during memory retrieval following auditory fear conditioning. These results suggest that auditory fear conditioning involves a close linkage between NMDA2A receptors and the CaMKII cascade.

    DOI

  • Nonphotic entrainment by 5-HT1A/7 receptor agonists accompanied by reduced Per1 and Per2 mRNA levels in the suprachiasmatic nuclei

    K Horikawa, S Yokota, K Fuji, M Akiyama, T Moriya, H Okamura, S Shibata

    JOURNAL OF NEUROSCIENCE   20 ( 15 ) 5867 - 5873  2000.08

     View Summary

    In mammals, the environmental light/dark cycle strongly synchronizes the circadian clock within the suprachiasmatic nuclei (SCN) to 24 hr. It is well known that not only photic but also nonphotic stimuli can entrain the SCN clock. Actually, many studies have shown that a daytime injection of 8-hydroxy-2-(di-n-propylamino) tetralin (8-OH DPAT), a serotonin 1A/7 receptor agonist, as a nonphotic stimulus induces phase advances in hamster behavioral circadian rhythms in vivo, as well as the neuron activity rhythm of the SCN in vitro. Recent reports suggest that mammalian homologs of the Drosophila clock gene, Period (Per), are involved in photic entrainment. Therefore, we examined whether phase advances elicited by 8-OH DPAT were associated with a change of Period mRNA levels in the SCN. In this experiment, we cloned partial cDNAs encoding hamster Per1, Per2, and Per3 and observed both circadian oscillation and the light responsiveness of Period. Furthermore, we found that the inhibitory effect of 8-OH DPAT on hamster Per1 and Per2 mRNA levels in the SCN occurred only during the hamster's mid-subjective day, but not during the early subjective day or subjective night. The present findings demonstrate that the acute and circadian time-dependent reduction of Per1 and/or Per2 mRNA in the hamster SCN by 8-OH DPAT is strongly correlated with the phase resetting in response to 8-OH DPAT.

  • The 5 ' upstream region of mPer1 gene contains two promoters and is responsible for circadian oscillation

    S Yamaguchi, S Mitsui, S Miyake, L Yan, H Onishi, K Yagita, M Suzuki, S Shibata, M Kobayashi, H Okamura

    CURRENT BIOLOGY   10 ( 14 ) 873 - 876  2000.07

     View Summary

    The mPer1 gene is assumed to be a key molecule in the regulation and functioning of the mammalian circadian clock, which is based on the oscillation generated by a transcription-(post)translation feedback loop of a set of clock genes [1]. Robust circadian oscillation and acute light-elicited induction of mPer1 mRNA expression have been observed in the suprachiasmatic nucleus (SCN), the mammalian circadian center [2,3]. To Investigate the mechanism underlying the complex regulation of mPer1 expression, we isolated and characterized the 5' upstream region of the mPer1 gene. Unexpectedly, we identified two promoters, each followed by alternative first exons of mPer1, Consistent with the presence of multiple E-boxes in the promoters, exon-specific in site hybridization of the SCN established that both promoters function in circadian oscillation and in light-induction of mPer1 expression. Transgenic mice carrying the 5' upstream region of the mPer1 gene fused to the luciferase gene demonstrated that a DNA fragment carrying both promoter regions is sufficient to elicit striking circadian oscillation in the SCN and responsiveness to light. Moreover, luminescence in the SCN accurately mirrored the mPer1 transcriptional activity. These transgenic mice will be very useful for monitoring clock-specific mPer1 expression in intact organisms and to follow the circadian clock in real time.

    DOI

  • Postnatal development of a GABA deficit and disturbance of neural functions in mice lacking GAD65

    O Stork, FY Ji, K Kaneko, S Stork, Y Yoshinobu, T Moriya, S Shibata, K Obata

    BRAIN RESEARCH   865 ( 1 ) 45 - 58  2000.05

     View Summary

    The 65-kDa isoform of glutamic acid decarboxylase (GAD65) is believed to play an essential role for GABA synthesis in the central nervous system. Using mice with targeted disruption of the GAD65 gene (GAD65(-/-) mice) we investigated the contribution of GAD65 to GABA synthesis in different brain areas during postnatal development and in adulthood. In the amygdala, hypothalamus and parietal cortex of GAD65(+/+) mice an increase of GABA levels was observed during postnatal development, most prominently between the first ii- mice and and second month after birth. This increase appeared to be dependent on GAD65, as it was delayed by 2 months in GAD65 was not observed in GAD65(-/-) mice. Likely as a consequence of their GABA deficit, adult GAD65(-/-) mice showed a largely abnormal neural activity with frequent paroxysmal discharges and spontaneous seizures. They furthermore displayed increased anxiety-like behaviour in a light/dark avoidance test and reduced intermale aggression, as well as a reduced forced-swimming-induced immobility indicative of an antidepressant-like behavioural change. Adult GAD65(-/-) mice did not show behavioural disturbances except for a reduced aggressive behaviour that was comparable to that in GAD65(-/-) mice. We conclude that GAD65-mediated GABA synthesis may be crucially involved in control of emotional behaviour and indispensable fur a tonic inhibition that prevents the development of hyperexcitability in the maturating central nervous system. Aggressive, and possibly other social behaviour may be especially prone to regulation through GAD65-mediated GABA synthesis. (C) 2000 Elsevier Science B.V. All rights reserved.

    DOI

  • PLC beta 4-independent Ca2+ rise via muscarinic receptors in the mouse suprachiasmatic nucleus

    M Ikeda, T Sugiyama, K Suzuki, T Moriya, S Shibata, M Katsuki, CN Allen, T Yoshioka

    NEUROREPORT   11 ( 5 ) 907 - 912  2000.04

     View Summary

    Cholinergic regulation of the suprachiasmatic nucleus (SCN) has been extensively studied although the intracellular signaling mechanisms are not well understood. We examined immunostaining for phospholipase C-beta (PLC-beta) families that couple to muscarinic acetylcholine receptors (mAChR) and demonstrated the expression of PLC-beta 1 and beta 4 in the mouse SCN. Ca2+ imaging analysis indicated that the MI-mAChR antagonist, pirenzepine blocked carbachol-induced Ca2+ elevation in the SCN and the response was equivalent between the wild type and the PLC-beta 4-knockout mice. In addition. the knockout mice displayed locomotor and temperature rhythms coupling to 24 h light/dark cycles. Therefore, it was proposed that PLC-beta 1 but not PLC-beta 4 was involved in the mAChR-mediated Ca2+ signaling in the SCN. NeoroReport 11:907-912 (C) 2000 Lippincott Williams & Wilkins.

  • Involvement of glial fibrillary acidic protein (GFAP) expressed in astroglial cells in circadian rhythm under constant lighting conditions in mice

    T Moriya, Y Yoshinobu, Y Kouzu, A Katoh, H Gomi, M Ikeda, T Yoshioka, S Itohara, S Shibata

    JOURNAL OF NEUROSCIENCE RESEARCH   60 ( 2 ) 212 - 218  2000.04

     View Summary

    To clarify the role of glial fibrillary acidic protein (GFAP)expressed glial cells in the circadian clock, we examined GFAP expression in the suprachiasmatic nucleus (SCN) and the intergeniculate leaflet (IGL) under various lighting conditions in mice. We demonstrated that GFAP expression did not show daily change in the SCN under a light-dark cycle; however, long-term housing under constant lighting conditions led to dramatic changes in GFAP expression, i.e., a decrease in the SCN and an increase in the IGL. Furthermore, mice that had a targeted deletion in the GFAP gene (GFAP mutant mice) showed longer and more arrhythmic circadian activity rhythms in constant lighting conditions than wild-type mice, while GFAP mutant mice exhibited stable circadian rhythms both in a light-dark cycle and constant darkness, and showed normal entrainment to environmental light stimuli. These results suggest that the GFAP-expressed astroglial cells in the SCN and the IGL may have some role in circadian oscillation under constant lighting conditions. (C) 2000 Wiley-Liss, Inc.

    DOI

  • Mutant mice lacking ryanodine receptor type 3 exhibit deficits of contextual fear conditioning and activation of calcium/calmodulin-dependent protein kinase II in the hippocampus

    Y Kouzu, T Moriya, H Takeshima, T Yoshioka, S Shibata

    MOLECULAR BRAIN RESEARCH   76 ( 1 ) 142 - 150  2000.03

     View Summary

    As it is known that ryanodine receptor type 3 is expressed in the hippocampus, we examined the contribution of this receptor to contextual fear conditioning behavior and to the activation of Ca2+/calmodulin-dependent protein kinase II using mice lacking the receptor. Ryanodine receptor type 3-deficient mice exhibited impairments of performance in the contextual fear conditioning test, passive avoidance test, and Y-maze learning test. Both the activities of Ca2+/calmodulin-dependent protein kinase II beta and Ca2+/calmodulin-dependent protein kinase II alpha were significantly increased in the experimental group compared to the control group in the hippocampus, but not in the cingulate cortex on the testing day 24 h after contextual fear training. However, the activities of Ca2+/calmodulin-dependent protein kinase II beta and alpha were almost the same in the experimental and control groups in the hippocampus on the training day. Ryanodine receptor type 3-deficient mice did not show the increment of Ca2+/calmodulin-dependent protein kinase II beta and alpha activities in the hippocampus on the testing day. In addition, these mutant mice showed the reduction of fear response in the elevated plus-maze test. The present results suggest that calcium-induced calcium release through the activation of ryanodine receptor type 3 in the hippocampus is important to the expression of the performance of contextual learning through the elevation of Ca2+/calmodulin-dependent protein kinase II beta and alpha activities. (C) 2000 Elsevier Science B.V. All rights reserved.

    DOI

  • 時計遺伝子とリズム同調

    守屋孝洋, 柴田重信

    神経研究の進歩   44   874 - 882  2000

    DOI

  • 時刻認知と体内時計

    柴田重信

    脳の科学   22   543 - 547  2000

  • 体内時計とくすり

    柴田重信

    脳21   3   101 - 107  2000

  • Involvement of ryanodine receptor type 3 in dopamine release from the striatum: Evidence from mutant mice lacking this receptor

    K Wan, T Moriya, M Akiyama, H Takeshima, S Shibata

    BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS   266 ( 2 ) 588 - 592  1999.12

     View Summary

    Although it is known that ryanodine receptor type 3 is expressed in the striatum, the function of this receptor has not been elucidated. Therefore, we examined whether caffeine- and ryanodine-induced dopamine release in striatal slices is affected in mice lacking ryanodine receptor type 3, Pretreatment with thapsigargin, an inhibitor of the Ca2+ ATPase pump of the endoplasmic reticulum, abolished caffeine- or ryanodine-induced dopamine release in slices from normal mice. Dopamine concentration in the striatum and KCl-induced dopamine release were unaffected by a ryanodine receptor type 3 deficiency. Ryanodine-induced dopamine release was significantly attenuated in mice lacking ryanodine receptor type 3, whereas caffeine-induced dopamine release was partially attenuated. Caffeine produced a similar hypermotor activity in both wild and homozygous mice. The present results suggest the involvement of ryanodine receptor type 3 in dopamine release from the striatum. (C) 1999 Academic Press.

    DOI

  • Modulation of mPer1 gene expression by anxiolytic drugs in mouse cerebellum

    M Akiyama, T Kirihara, S Takahashi, Y Minami, Y Yoshinobu, T Moriya, S Shibata

    BRITISH JOURNAL OF PHARMACOLOGY   128 ( 7 ) 1616 - 1622  1999.12

     View Summary

    1 The mPer1 and mPer2 genes are putative mouse clock genes that regulate circadian oscillator present in the suprachiasmatic nucleus (SCN) neuron. While they are also expressed in the granular cell layer in the cerebellum, their function is unknown. In a first step to verify the physiological roles of mPer1 and mPer2 genes in the cerebellum, we examined the effects of benzodiazepines on the expression of the mPer1 and mPer2 genes.
    2 mPer2 mRNA expression was higher at ZT16 than ZT4 in the mouse cerebellum.
    3 High-dose administration of diazepam (10 mg kg(-1)) or triazolam (1 mg kg(-1)) reduced mPer1 mRNA level 1 h after treatment in the cerebellum.
    4 Reduced expression of mPer1 by diazepam treatment was transient. No difference of mPer1 mRNA level between diazepam (10 mg kg(-1))- and vehicle-treated group was observed 6 h after treatment.
    5 Administration of high doses of tandospirone (30 mg kg(-1)), a non-benzodiazepine anxiolytic also reduced mPer1 mRNA expression 1 h after treatment.
    6 Administration of high doses of clozapine (5 mg kg(-1)) or haloperidol (1 mg kg(-1)) impaired the rota-rod performance without affecting on mPer1 mRNA level.
    7 Diazepam and tandospirone inhibited the expression of Per1 mRNA in the primary cultured cerebellum granule cells.
    8 Transient reductions of mPer1 mRNA levels by various benzodiazepines and tandospirone is associated with impairment of coordinated movement, such as rota-rod performance and equilibrium.

    DOI

  • Sensitization of methamphetamine-induced disorganization of daily locomotor activity rhythm in male rats

    T Nikaido, T Moriya, R Takabayashi, M Akigama, S Shibata

    BRAIN RESEARCH   845 ( 1 ) 112 - 116  1999.10

     View Summary

    Methamphetamine (MAP) was administered to rats through drinking water repeatedly (three sessions, one session:administration for 60 days followed by withdrawal of 30 days) in order to examine whether or not MAP-induced disorganization of daily activity rhythm is sensitized. Each session (60 days) was divided into six blocks of 10 days. In the Ist session, daily locomotor activity rhythm of rats became disorganized around at 40 days (4th block) after the start of MAP drinking, However, MAP-induced disorganization of daily activity rhythm appeared at 20 days (2nd block) in the 2nd session and at 10 days (Ist block) in the 3rd session following re-start of MAP drinking. On the other hand, the amount of MAP intake was decreased on the 2nd and 3rd sessions as compared with the Ist session. These results indicate that the mechanism of MAP-induced disorganization of daily activity rhythm may involve sensitization. (C) 1999 Elsevier Science B.V. All rights reserved.

    DOI

  • Involvement of glutamate release in substance P-induced phase delays of suprachiasmatic neuron activity rhythm in vitro

    T Hamada, S Yamanouchi, A Watanabe, S Shibata, S Watanabe

    BRAIN RESEARCH   836 ( 1-2 ) 190 - 193  1999.07

     View Summary

    The suprachiasmatic nucleus (SCN) has been identified as a mammalian circadian rhythm clock. Treatment with substance P (SP) at zeitgeber time 13-14 produced phase delays of circadian rhythm in spontaneous neural activity in SCN neurons in vitro. SP-induced phase delays are blocked by treatment with not only SP receptor antagonist, spantide, but N-methyl-D-aspartate receptor antagonist, MK-801. In the biochemical experiment, we demonstrated that SP-induced glutamate release from the SCN slices was observed by the high-performance liquid chromatography assay. The present results suggest that glutamate release may be involved in SP-induced phase delays. (C) 1999 Elsevier Science B.V. All rights reserved.

    DOI

  • 心身の開放と脳内物質

    柴田重信

    体育の科学   49 ( 6月 ) 448 - 452  1999.06

  • Effects of prolyl endopeptidase inhibitors and neuropeptides on delayed neuronal death in rats

    Y Shishido, M Furushiro, S Tanabe, S Shibata, S Hashimoto, T Yokokura

    EUROPEAN JOURNAL OF PHARMACOLOGY   372 ( 2 ) 135 - 142  1999.05

     View Summary

    We investigated the effects of the prolyl endopeptidase inhibitors 1-[1-(Benzyloxycarbonyl)-L-prolyl]prolinal (Z-Pro-Prolinal) and N-benzyloxycarbonyl-thioprolyl-thioprolinal-dimethylacetal (ZTTA) on delayed neuronal death induced by four-vessel-occlusion transient ischemia in rats. We also examined the effects of [pGlu(4), Cyt(6), Arg(8)]vasopressin (vasopressin-(4-9)) and thyrotropin-releasing hormone (TRH) on the delayed neuronal death. Furthermore, we investigated the role of vasopressin receptors in the effects of vasopressin and prolyl endopeptidase inhibitors. Z-Pro-Prolinal, vasopressin-(4-9) and TRH protected pyramidal cells in the CA1 subfield of the rat hippocampus from delayed neuronal death after 10-min ischemia, The effect of vasopressin-(4-9) was abolished by vasopressin receptor antagonists. The effect of Z-Pro-Prolinal was also abrogated by the antagonists. These results suggest that the neuroprotective effect of prolyl endopeptidase inhibitors is mediated by neuropeptides such as [Arg(8)]vasopressin and TRH, and indicate the involvement of vasopressin receptors in the neuroprotective effect of vasopressin-(4-9) and prolyl endopeptidase inhibitors. (C) 1999 Elsevier Science B.V. All rights reserved.

    DOI

  • ZTTA, a prolyl endopeptidase inhibitor, potentiates the arginine-vasopressirn-induced incorporation of [C-14]leucine in rat amygdaloid and cortical slices

    Y Shishido, T Tanaka, S Tanabe, M Furushiro, S Hashimoto, T Yokokura, S Shibata, S Watanabe

    PHARMACEUTICAL RESEARCH   16 ( 3 ) 463 - 465  1999.03

    DOI

  • Diurnal regulation of a DNA binding protein to the period repeat sequence in the SCN nuclear extract of rat brain

    T Hamada, K Kako, H Wakamatsu, S Shibata, S Watanabe, N Ishida

    MOLECULAR BRAIN RESEARCH   65 ( 2 ) 211 - 215  1999.03

     View Summary

    We previously reported the mammalian period repeat mRNA fluctuates during circadian time in the rat suprachiasmatic nucleus (SCN) which is considered to be a clock pacemaker in mammalian brain. Presently we discovered a period repeat sequence (PR) DNA-binding protein in the rat SCN nuclear extract. In the SCN, the binding activity of PR DNA-binding protein to (ACAGGC)(3) was most highest during the late day and most lowest during the late night by electro-mobility shift assay (EMSA). In the cortex nuclear extract, the binding of PR DNA-binding protein did not show a significant variation during a day. This is the first report to show the existence of diurnal regulated PR DNA-binding protein in the SCN. (C) 1999 Elsevier Science B.V. All rights reserved.

    DOI

  • Inhibition of light- or glutamate-induced mPer1 expression represses the phase shifts into the mouse circadian locomotor and suprachiasmatic firing rhythms

    M Akiyama, Y Kouzu, S Takahashi, H Wakamatsu, T Moriya, M Maetani, S Watanabe, H Tei, Y Sakaki, S Shibata

    JOURNAL OF NEUROSCIENCE   19 ( 3 ) 1115 - 1121  1999.02

     View Summary

    mPer1, a mouse gene, is a homolog of the Drosophila clock gene period and has been shown to be closely associated with the light-induced resetting of a mammalian circadian clock. To investigate whether the rapid induction of mPer1 after light exposure is necessary for light-induced phase shifting, we injected an antisense phosphotioate oligonucleotide (ODN) to mPer1 mRNA into the cerebral ventricle. Light-induced phase delay of locomotor activity at CT16 was significantly inhibited when the mice were pretreated with mPer1 antisense ODN 1 hr before light exposure. mPer1 sense ODN or random ODN treatment had little effect on phase delay induced by light pulses. In addition, glutamate-induced phase delay of suprachiasmatic nucleus (SCN) firing rhythm was attenuated by pretreatment with mPer1 antisense ODN, but not by random ODN. The present results demonstrate that induction of mPer1 mRNA is required for light- or glutamate-induced phase shifting, suggesting that the acute induction of mPer1 mRNA in the SCN after light exposure is involved in light-induced phase shifting of the overt rhythm.

  • 体内時計と時間薬理学

    柴田重信

    日本薬剤師会雑誌   51 ( 12 ) 1879 - 1885  1999

  • 体内時計の分子基盤研究の競争

    柴田重信

    日本薬理学雑誌   114   242  1999

  • 体内時計とセロトニン

    堀川和政, 柴田重信

    Molecular Medicine   36   1143 - 1149  1999

  • 概日リズムと細胞内信号系

    守屋孝洋, 柴田重信

    生体の科学   50   200 - 206  1999

    DOI

  • 生体リズムと薬物

    柴田重信

    日本神経精神薬理学雑誌   19   11 - 20  1999

  • 創薬の視点に立った生体リズム

    柴田重信

    月刊薬事   41   214 - 217  1999

  • Melatonin normalizes the reentrainment of senescence accelerated mice (SAM) to a new light-dark cycle

    S Shibata, M Asai, Oshima, I, M Ikeda, T Yoshioka

    MELATONIN AFTER FOUR DECADES   460   261 - 270  1999

    DOI

  • Diurnal DNA binding activity isolated from rat brain clock pacemaker recognize period repetitive sequence in vitro.

    Hamada;T, Kako;K, Wakamatsu;H, Suzuki;S, Takeuchi;J, Shibata;S, and, Ishida;N, 加香, 孝一郎

    Brain Research Molecular Brain Research   65   211-215  1999.01  [Refereed]

  • Effect of ZTTA, a prolyl endopeptidase inhibitor, on memory impairment in a passive avoidance test of rats with basal forebrain lesions

    Y Shishido, M Furushiro, S Tanabe, A Taniguchi, S Hashimoto, T Yokokura, S Shibata, T Yamamoto, S Watanabe

    PHARMACEUTICAL RESEARCH   15 ( 12 ) 1907 - 1910  1998.12

    DOI

  • NMDA induced glutamate release front the suprachiasmatic nucleus: an in vitro study in the rat

    T Hamada, R Sonoda, A Watanabe, M Ono, S Shibata, S Watanabe

    NEUROSCIENCE LETTERS   256 ( 2 ) 93 - 96  1998.11

     View Summary

    The suprachiasmatic nucleus (SCN) has been identified as a pacemaker for mammalian circadian rhythms. Excitatory amino acid receptors, especially N-methyl-D-aspartate (NMDA) receptors, have been considered to play an important role in the transmission of light information from the retina to the circadian clocks in the SCN. In the present study, we showed that application of NMDA at circadian time (CT) 12-15 induced significant glutamate release from the SCN region in vitro. The NMDA-induced glutamate release was blocked by co-application of the NMDA receptor antagonist MK-801, but not by that of tetrodotoxin. These results suggested that glutamate stimulated its own release by activating NMDA receptors. This NMDA-induced glutamate release through NMDA receptor-mediated mechanisms might be involved in NMDA-induced potent phase shifts. (C) 1998 Elsevier Science Ireland Ltd. All rights reserved.

    DOI

  • Potentiating action of MKC-242, a selective 5-HT1A receptor agonist, on the photic entrainment of the circadian activity rhythm in hamsters

    T Moriya, Y Yoshinobu, M Ikeda, S Yokota, M Akiyama, S Shibata

    BRITISH JOURNAL OF PHARMACOLOGY   125 ( 6 ) 1281 - 1287  1998.11

     View Summary

    1 Serotonergic projections from the midbrain raphe nuclei to the suprachiasmatic nuclei (SCN) are known to regulate the photic entrainment of circadian clocks. However, it is not known which 5-hydroxytryptamine (5-HT) receptor subtypes are involved in the circadian regulation. In order to verify the role of 5-HT1A receptors, we examined the effects of 5-{3-[((2S)-1,4-benzodioxan-2-ylmethyl)amino]propoxy}-1,3-benzodioxole HCl (MKC-242), a selective 5-HT1A receptor agonist, on photic entrainment of wheel-running circadian rhythms of hamsters.
    2 MKC-242 (3 mg kg(-1), i.p.) significantly accelerated the re-entrainment of wheel-running rhythms to a new 8 h delayed or advanced light-dark cycle.
    3 MKC-242 (3 mg kg(-1), i.p.) also potentiated the phase advance of the wheel-running rhythm produced by low (5 lux) or high (60 lux) intensity light pulses. In contrast, 8-hydroxydipropylaminotetralin (8-OH-DPAT)(5 mg kg(-1), i.p.), a well known 5-HT1A/5-HT7 receptor agonist, only suppressed low intensity (5 lux) light-induced phase advances.
    4 The potentiating actions of MKC-242 on light pulse-induced phase advances were observed even when injected 20 or 60 min after the light exposure.
    5 The potentiating action of MKC-242 was antagonized by WAY100635, a selective 5-HT1A receptor blocker, but not by ritanserin, a 5-HT2/5-HT7 receptor blocker, indicating that MKC-242 is activating 5-HT1A receptors.
    6 Light pulse-induced c-fas expression in the SCN and the intergeniculate leaflet (IGL) were unaffected by MKC-242 (3 mg kg(-1), i.p.).
    7 HPLC analysis demonstrated that MKC-242 (3 mg kg(-1), i.p.) decreased the 5-HIAA content in the SCN.
    8 The present results suggest that presynaptic 5-HT,A receptor activation may be involved in the potentiation of photic entrainment by MKC-242 in hamsters.

    DOI

  • Vulnerability of synaptic plasticity in the striatum of methamphetamine-sensitized rats

    Ikumi Arai, Takao Shimazoe, Akiko Yoshimatsu, Hirotaka Inoue, Shigenobu Shibata, Shigenori Watanabe

    Japanese Journal of Pharmacology   78 ( 1 ) 105 - 108  1998.09  [Refereed]

     View Summary

    We examined the influence of ischemia on methamphetamine (MAP)-induced behavioral sensitization and enhancement of dopamine (DA) release. After the recovery period of the ischemia operation, rats were treated with MAP (1 mg/kg, i.p.) once daily for 6 consecutive days. Re-administration of MAP (0.5 mg/kg, i.p.) potentiated the increase of locomotor activity after a 3-day withdrawal and the enhancement of DA release from striatal slices after a 6-day withdrawal. The MAP-induced sensitization was impaired by 5 min ischemia. On the other hand, the increase of locomotor activity induced by single MAP (1 mg/kg, i.p.) administration was impaired by 20 min of ischemia. Moreover, in saline-treated rats the increase of DA release from striatal slices induced by MAP (10 μM) application was also impaired by 20 min of ischemia. These results indicate that the neuronal plastic change may be very vulnerable to ischemia in MAP-induced sensitization.

    DOI PubMed

  • Methylcobalamin amplifies melatonin-induced circadian phase shifts by facilitation of melatonin synthesis in the rat pineal gland

    M Ikeda, M Asai, T Moriya, M Sagara, S Inoue, S Shibata

    BRAIN RESEARCH   795 ( 1-2 ) 98 - 104  1998.06

     View Summary

    Effects of methylcobalamin (methyl-B-12), a putative drug for treating human circadian rhythm disorders, on the melatonin-induced circadian phase shifts were examined in the rat. An intraperitoneal injection of 1-100 mu g/kg melatonin 2-h before the activity onset time (CT 10) induced phase advances of free-running activity rhythms in a dose-dependent manner (ED50 = 1.3 mu g/kg). Injection of methyl-B-12 (500 mu g/kg) prior to melatonin (1 mu g/kg) injection induced larger phase advances than saline preinjected controls, while the injection of methyl-B-12 in combination with saline did not induce a phase advance. These results indicate amplification of melatonin-induced sphase advances by methyl-B-12. Pinealectomy abolished the phase alternating effect of methyl-B-12, suggesting a site of action within the lineal gland. In fact, methyl-B-12 significantly increased the content of melatonin in the pineal collected 2-h after activity onset (CT 14). In contrast, no difference in melatonin content was found at CT 10, indicating that the effect of methyl-B-12 may be gated after the activity onset time when endogenous melatonin synthesis is known to increase. These results suggest that methyl-B-12 amplifies melatonin-induced phase advances via an increase in melatonin synthesis during the early subjective night at a point downstream from the clock regulation. (C) 1998 Elsevier Science B.V. All rights reserved.

    DOI

  • 記憶と生物時計

    柴田重信

    科学   68 ( 2 ) 160 - 164  1998.02

  • 光とメラトニン

    柴田重信

    ファルマシア   34   45 - 49  1998

  • 母児概日リズムの自律性と相互関係

    三宅良明, 長岡理明, 内藤成美, 大西美也子, 田中均, 佐藤和雄, 村井一郎, 柴田重信, 渡辺恭良, 大塚邦明

    組織培養工学   24   19 - 23  1998

  • 生体時計の生理学的,薬理学的,分子生物学的解析

    秋山正志, 守屋孝洋

    日本薬理学雑誌   112   243 - 250  1998

    DOI

  • N-methyl-D-aspartate receptors are indispensable for the formation of long-term potentiation in the rat suprachiasmatic nucleus in vitro

    Y Nisikawa, T Shimazoe, S Shibata, S Watanabe

    BRAIN RESEARCH   782 ( 1-2 ) 303 - 305  1998.01

     View Summary

    Optic nerve (ON) stimulation caused a postsynaptic field potential in the suprachiasmatic nucleus (SCN) of rat hypothalamic slices. The postsynaptic field potential was suppressed by 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX), a non-NMDA receptor antagonist, in a concentration-dependent manner, but not affected by D-amino-5-phosphonovaleric acid (APV), a competitive NMDA receptor antagonist. Tetanic stimulation to the ON induced long-term potentiation (LTP) in the SCN. Application of APV at 50 mu M inhibited the induction of LTP by tetanic stimulation but CNQX at lower dose (5 mu M) didn't inhibit it. These results suggest that NMDA receptors are indispensable for the induction of LTP after tetanic stimulation. (C) 1998 Elsevier Science B.V.

    DOI

  • Light-induced resetting of a mammalian circadian clock is associated with rapid induction of the mPer1 transcript

    Y Shigeyoshi, K Taguchi, S Yamamoto, S Takekida, L Yan, H Tei, T Moriya, S Shibata, JJ Loros, JC Dunlap, H Okamura

    CELL   91 ( 7 ) 1043 - 1053  1997.12

     View Summary

    To understand how light might entrain a mammalian circadian clock, we examined the effects of light on mPer1, a sequence homolog of Drosophila per, that exhibits robust rhythmic expression in the SCN.mPer1 is rapidly induced by short duration exposure to light at levels sufficient to reset the clock, and dose-response curves reveal that mPer1 induction shows both reciprocity and a strong correlation with phase shifting of the overt rhythm. Thus, in both the phasing of dark expression and the response to light mPer1 is most similar to the Neurospora clock gene fro. Within the SCN there appears to be localization of the induction phenomenon, consistent with the localization of both light-sensitive and light-insensitive oscillators in this circadian center.

    DOI

  • Concurrent blockade of beta-adrenergic and muscarinic receptors suppresses synergistically long-term potentiation of population spikes in the rat hippocampal CA1 region

    M Kobayashi, M Ohno, S Shibata, T Yamamoto, S Watanabe

    BRAIN RESEARCH   777 ( 1-2 ) 242 - 246  1997.11

     View Summary

    The muscarinic acetylcholine receptor antagonist scopolamine, but not the beta-adrenoceptor antagonist propranolol or atenolol, suppressed tetanus-induced long-term potentiation (LTP) of population spikes in the rat hippocampal CA1 region. When scopolamine was coapplied with propranolol or atenolol, a synergistic effect in preventing LTP generation was observed, On the other hand, the coapplication of scopolamine and atenolol failed to affect tetanus-induced LTP of field EPSP. These findings suggest that cooperative mechanisms via muscarinic and beta-adrenergic receptor activation might contribute to LTP induction in terms of the EPSP-spike potentiation, i.e., an increase in the excitability of hippocampal CA1 pyramidal cells after tetanic stimulation, but are independent of the tetanus-evoked potentiation of a synaptic component. (C) 1997 Elsevier Science B.V.

    DOI

  • Decreased level of light-induced Fos expression in the suprachiasmatic nucleus of diabetic rats

    S Yamanouchi, T Shimazoe, S Nagata, T Moriya, M Maetani, S Shibata, S Watanabe, K Miyasaka, A Kono, A Funakoshi

    NEUROSCIENCE LETTERS   227 ( 2 ) 103 - 106  1997.05

     View Summary

    We assessed light-induced Fos-immunoreactive cells in the suprachiasmatic nucleus of diabetic rats. The number of Fos-immunoreactive cells significantly decreased in diabetic Otsuka Long-Evans Tokushima Fatty (OLETF) rats as compared with control Long-Evans Tokushima Otsuka (LETO) rats. In contrast there was no decrease in the number of Fos-immunoreactive cells in young OLETF rats which have not yet developed diabetes. Two months after the administration of streptozotocin (STZ) to Wistar rats, the number of Fos-immunoreactive cells significantly decreased, although 1 week after the administration of STZ, the number had not yet changed in these STZ-induced diabetic rats. These results suggest that chronic diabetic (hyperglycemic) conditions may affect the light entraining responses in the suprachiasmatic nucleus (SCN). (C) 1997 Elsevier Science Ireland Ltd.

    DOI

  • The involvement of calmodulin and Ca2+/calmodulin-dependent protein kinase II in the circadian rhythms controlled by the suprachiasmatic nucleus

    T Fukushima, T Shimazoe, S Shibata, A Watanabe, M Ono, T Hamada, S Watanabe

    NEUROSCIENCE LETTERS   227 ( 1 ) 45 - 48  1997.05

     View Summary

    We investigated the involvement of calmodulin and Ca2+/calmodulin-dependent protein kinase II (CaMKII) in the photic entrainment of circadian rhythms using calmodulin inhibitors such as calmidazolium (CMZ) and trifluoperazine (TFP), and a CaMKII inhibitor, KN-62, in rats. Fos expression in the suprachiasmatic nucleus (SCN) of rats induced by photic stimulation (300 lux, 1 h) during the early subjective night of the rats was inhibited by treatment with CMZ (10 mg/kg i.p.) or TFP (20 mg/kg i.p.) 30 min before photic stimulation. With respect to the neuronal firing rate in the rat SCN slice, KN-62 and CMZ application during the early subjective night attenuated the glutamate (10 mu M)-induced phase shift. The present results suggest that calmodulin and CaMKII are involved in the photic entrainment mechanism in the rodent SCN. (C) 1997 Elsevier Science Ireland Ltd.

    DOI

  • Z-321, a prolyl endopeptidase inhibitor, augments the potentiation of synaptic transmission in rat hippocampal slices

    N Miura, S Shibata, S Watanabe

    BEHAVIOURAL BRAIN RESEARCH   83 ( 1-2 ) 213 - 216  1997.02

     View Summary

    The present study investigated the effects of arginine-vasopressin (AVP) and (1-[3-(2-indanylacetyl)-L-thioprolyl] pyrrolidine (Z-321), an inhibitor of prolyl endopeptidase (PEP; (EC 3.4.21.26)) which degrades AVP in vitro, on the short-lasting potentiation of the field excitatory postsynaptic potentials (EPSP) coupled with a weak tetanus. The EPSP, after the electrical stimulation of the Schaffer collateral/commissural pathway, were recorded in the CA1 region of rat hippocampal slices. AVP at 10(-8) M and Z-321 at 10(-4) M augmented the potentiation induced by the weak tetanus; the magnitude of the post-tetanic potentiation of the EPSP was enhanced and the potentiation lasted for 60 min. In contrast, the racemic D-thioprolyl compound of Z-321, which virtually lacks any inhibitory effects on PEP, failed to affect the potentiation at 10(-4) M. The facilitatory effect of Z-321 was reversed by the application of [d(CH2)(5),Tyr(Me)(2)]AVP (10(-8) M), an antagonist of the AVP V-1 receptors, indicating that the effect of Z-321 was mediated through the V-1 receptors. These findings suggest that Z-321 augmented the potentiation due to its inhibitory influence on the AVP degradation by PEP.

    DOI

  • 視交叉上核におけるリズム発振と同調の分子機構

    秋山正志, 柴田重信

    日本時間生物学会会誌   4   22 - 32  1997

  • 概日リズムとセロトニン神経系

    池田真行, 柴田重信

    Molecular Medicine   34   286 - 293  1997

  • 視交叉上核におけるNMDA受容体と長期増強現象

    守屋孝洋, 柴田重信

    ブレインサイエンス   8 ( 1 ) 49 - 57  1997

  • Methamphetamine-induced sensitization of dopamine release via a metabotropic glutamate receptor mediated pathway in rat striatal slices

    Ikumi Arai, Takao Shimazoe, Shigenobu Shibata, Hirotaka Inoue, Akiko Yoshimatsu, Shigenori Watanabe

    Japanese Journal of Pharmacology   73 ( 3 ) 243 - 246  1997  [Refereed]

     View Summary

    We studied the roles of metabotropic glutamate receptors in methamphetamine (MAP)-induced sensitization of dopamine (DA) release from striatal slices. Rats were first treated with MAP (1 mg/kg, i.p.) once daily for 6 consecutive days. After a 6-day withdrawal, DA release from striatal slices evoked by ±-1-aminocyclopentane-trans-1,3-dicarboxylic acid (trans-ACPD) was measured. trans-ACPD-induced DA release was significantly enhanced in MAP-sensitized rats, but the inactive form of trans-ACPD (1R,3S-ACPD) did not enhance DA release. The active form of trans-ACPD (1S,3R-ACPD) (0.1 mM)-evoked DA release was attenuated by treatment with 0.4 mM RS-α-methyl-4-carboxyphenylglycine, a metabotropic glutamate receptor antagonist. The present results suggest that metabotropic glutamate receptors play an important role in expression of MAP-induced sensitization.

    DOI PubMed

  • Methylcobalamin induces a long-lasting enhancement of the field potential in rat suprachiasmatic nucleus slices

    Yukiko Nishikawa, Shigenobu Shibata, Takao Shimazoe, Shigenori Watanabe

    Neuroscience Letters   220 ( 3 ) 199 - 202  1996.12  [Refereed]

     View Summary

    Optic nerve stimulation has been reported to evoke a field potential (FP) in rat suprachiasmatic nucleus (SCN) slices. Methylcobalamin,δ-(5,6-dymethylbenzimidazolyl)-Co-methyl-cobamide (Me-B12) enhanced this FP and the enhancement lasted more than 1 h after washing out. Maximal enhancement (143.6 ± 9.8%) was achieved at a concentration of 10 μM. By contrast, cyanocobalamin containing CN- instead of CH3- showed no enhancement of the amplitude in the FP. Me-B12 induced enhhncement of FP was strongly blocked by an N-methyl-D-aspartate (NMDA) receptor antagonist, DL-2-amino-5-phosphonovaleric acid (APV). These results indicate that CH3- in the Me-B12 is required to modulate the FP amplitude and the NMDA receptor is involved in the long-lasting FP enhancement induced by Me-B12. The present results suggest that Me-B12 modifies the photic entrainment of the circadian clock in the suprachiasmatic nucleus via an activation of NMDA receptors.

    DOI PubMed

  • Involvement of 5-HT1A receptor mechanisms in the inhibitory effects of methamphetamine on photic responses in the rodent suprachiasmatic nucleus

    T Moriya, S Yamanouchi, T Fukushima, T Shimazoe, S Shibata, S Watanabe

    BRAIN RESEARCH   740 ( 1-2 ) 261 - 267  1996.11

     View Summary

    We examined the role of serotonin IA (5-HT1A) receptors in the inhibitory effects of methamphetamine (MA) on photic entrainment to the circadian pacemaker in the suprachiasmatic nucleus (SCN) of rodents. MA inhibited optic nerve stimulation-evoked field potential in the SCN, light-induced Fos expression in the SCN and light-induced phase shift of hamster wheel-running rhythm. NAN-190, a 5-HT1A receptor antagonist, eliminated the inhibitory effects of MA. NAN-190 has also been reported to antagonize alpha 1 adrenergic receptors. However, prazosin, which selectively antagonizes alpha(1) adrenergic receptors, did not affect the inhibitory action of RIA on light-induced Fos expression. In addition, parachloroamphetamine, which is known to be a 5-HT releaser, dose-dependently inhibited light-induced phase shift of wheel-running rhythm. These findings suggest that elevation of endogenous 5-HT levels by RIA inhibits the photic entraining responses of the circadian pacemaker in the SCN via 5-HT1A receptor stimulation of the 5-HT released by MA.

    DOI

  • Adenosine A1-receptor agonist attenuates the light-induced phase shifts and fos expression in vivo and optic nerve stimulation-evoked field potentials in the suprachiasmatic nucleus in vitro

    A Watanabe, T Moriya, Y Nisikawa, T Araki, T Hamada, S Shibata, S Watanabe

    BRAIN RESEARCH   740 ( 1-2 ) 329 - 336  1996.11

     View Summary

    Adenosine is widely accepted to act as an inhibitory neuromodulator in the mammalian central nervous system. In the present study, we examined whether adenosine receptor agonist modifies the photic entraining responses in the rat suprachiasmatic nucleus both in vivo and in vitro. Light (200 lux, 15 min)-induced phase shifts of hamster wheel-running rhythms was attenuated by a systemic administration of AI-adenosine receptor agonist N-6-cyclohexyladenosine (N-CHA) in a dose-dependent manner; 0.5 mg/kg N-CHA caused 60% inhibition of light-induced phase shifts. On the other hand, A2-adenosine receptor agonist N-6-[2-(3,5-dimethoxyphenyl)-2-(2-methylphenyl)-ethyl]adenosine (DPMA) failed to inhibit light-induced phase shifts. Systemic administration of N-CHA but not of DPMA inhibited light (300 lux, 1 h)-induced Fos expression in the suprachiasmatic nucleus in a dose-dependent manner; 1 mg/kg N-CHA caused 73% inhibition of light-induced Fos expression. Bath application of N-CHA but not of DPMA inhibited optic nerve stimulation-evoked field potentials in rat suprachiasmatic nucleus slices. The present results suggest that activation of adenosine Al-receptor attenuates the photic input through the inhibition of retinohypotalamic pathway to the SCN.

    DOI

  • Enhancement of dopamine release from the striatum through metabotropic glutamate receptor activation in methamphetamine sensitized rats

    Arai, I, T Shimazoe, S Shibata, H Inoue, A Yoshimatsu, S Watanabe

    BRAIN RESEARCH   729 ( 2 ) 277 - 280  1996.08

     View Summary

    An intracerebral microdialysis technique was applied to study the effect of metabotropic glutamate receptor (mGluR) agonist on dopamine release in the striatum of methamphetamine (MAP)-sensitized rats. Rats were treated with MAP (1 mg/kg, i.p.) once daily for 6 consecutive days, followed by a 6-day withdrawal. Perfusion of 0.1 mM (1S:3R)-1-aminocyclopentane-trans-1,3-dicarboxylic acid through a microdialysis probe placed in the striatum enhanced the extracellular dopamine level, and induced stereotyped behavior in MAP-sensitized rats. The enhancement of dopamine release and the stereotyped behavior were attenuated by co-perfusion of 0.4 mM RS-alpha-methyl-4-carboxyphenyl-glycine, a mGluR antagonist. The present results sus est that mGluRs may be involved in the expression of MAP-induced sensitization.

    DOI

  • N-G-nitro-L-arginine methyl ester attenuates the maintenance and expression of methamphetamine-induced behavioral sensitization and enhancement of striatal dopamine release

    H Inoue, Arai, I, S Shibata, S Watanabe

    JOURNAL OF PHARMACOLOGY AND EXPERIMENTAL THERAPEUTICS   277 ( 3 ) 1424 - 1430  1996.06

     View Summary

    We examined the roles of nitric oxide (NO) in methamphetamine (MAP)-induced behavioral sensitization and enhancement of striatal dopamine (DA) release using both in vivo and in vitro methods. Repeated administration of MAP produced augmentation of MAP-induced locomotor activity after 3-day withdrawal of MAP and an enhancement of MAP-evoked DA release from striatal slices after 6-day withdrawal. When the NO synthase (NOS) inhibitor N-G-nitro-L-arginine methyl ester (L-NAME) was administered only during the period of MAP withdrawal, the behavioral sensitization and enhancement of DA release were attenuated significantly. In contrast, N-G-nitro-D-arginine methyl ester, an inactive isomer of L-NAME, exhibited no such effect. When L-NAME was administered acutely before the challenge injection of MAP, behavioral sensitization was also attenuated only when the dose of L-NAME was high. Coadministration of L-NAME with MAP did not block the development of sensitization to MAP. We also examined whether MAP-induced behavioral sensitization and enhancement of DA release could be observed in vivo in a microdialysis experiment. Challenge injection of MAP caused marked enhancement of DA release in MAP-sensitized rats compared with saline-treated controls corresponding to robust augmentation of locomotor activity. When L-NAME was injected during the MAP withdrawal period, the enhancement of DA release and locomotor activity induced by challenge injection of MAP were attenuated. These results suggest that NO production plays a role in the maintenance (expression) of MAP-induced behavioral sensitization and enhancement of DA release but not in the development of these effects.

  • Positron-labeled anitoxidant 6-deoxy-6-[F-18]fluoro-L-ascorbic acid: Increased uptake in transient global ischemic rat brain

    F Yamamoto, S Shibata, S Watanabe, K Masuda, M Maeda

    NUCLEAR MEDICINE AND BIOLOGY   23 ( 4 ) 479 - 486  1996.05

     View Summary

    The in vivo uptake and distribution of 6-deoxy-6-[F-18]fluoro-L-ascorbic acid (F-18-DFA) were investigated in rat brains following postischemic reperfusion. Global cerebral ischemia was in induced in male Wistar rats for 20 min by occlusion of four major arteries. Two time points were chosen for F-18-DFA injection to rats subjected to cerebral ischemia, at the start of recirculation and 5 days following recirculation. The rats were then killed at 2 h after tail vein administration of F-18-DFA and tissue radioactivity concentration was determined. Increased uptake of radioactivity in particular brain regions, including the cerebral cortex, hypothalamus, and amygdala following injection of F-18-DFA, compared to the sham operated control, was observed 5 days after reperfusion. Similar results were also obtained in in vitro experiments using brain slices. Abnormal in vivo accumulation of Ca-45, a marker of regional postischemic injury, was observed in these brain regions in tissue dissection experiments. Furthermore, metabolite analysis of nonradioactive DFA using F-19-NMR showed that DFA remained intact in the postischemic reperfusion brain. The present results indicate that F-18-DFA increasingly accumulates in damaged regions of postischemic reperfusion brain.

    DOI

  • Methamphetamine modifies the photic entraining responses in the rodent suprachiasmatic nucleus via serotonin release

    M Ono, A Watanabe, Y Matsumoto, T Fukushima, Y Nishikawa, T Moriya, S Shibata, S Watanabe

    NEUROSCIENCE   72 ( 1 ) 213 - 224  1996.05

     View Summary

    We examined whether methamphetamine modifies the photic entraining responses in the rat suprachiasmatic nucleus. Optic nerve stimulation increased vasoactive intestinal polypeptide release from rat suprachiasmatic nucleus slices, and methamphetamine inhibited this increase in a concentration-dependent manner. Optic nerve stimulation has been reported to evoke field potentials in rat suprachiasmatic nucleus slices. Methamphetamine attenuated this held potential, and maximal inhibition (75.5%) was achieved at a concentration of 100 mu M. Systemic administration of methamphetamine (1-5 mg/kg) inhibited light (300 lux, 1 h)-induced Fos expression in the suprachiasmatic nucleus; methamphetamine at a dose of 5 mg/kg, i.p. caused 40% inhibition of light-induced Fos expression. We examined whether the inhibitory effect of methamphetamine on photic entraining responses mediates serotonin release from the suprachiasmatic nucleus. High-performance liquid chromatographic analysis revealed that methamphetamine application increased serotonin release from rat suprachiasmatic nucleus slices in a concentration-dependent manner, but did not affect noradrenaline release. In addition, reduction of serotonin content attenuated the effect of methamphetamine on field potential induced by optic nerve stimulation in vitro and also light-induced phase advances of wheel running activity rhythm in vivo.
    The present results support the idea that methamphetamine produces an inhibitory effect on photic entrainment in the suprachiasmatic nucleus via serotonin release.

    DOI

  • Chronic administration of methamphetamine does not affect the suprachiasmatic nucleus-operated circadian pacemaker in rats

    T Moriya, T Fukushima, T Shimazoe, S Shibata, S Watanabe

    NEUROSCIENCE LETTERS   208 ( 2 ) 129 - 132  1996.04

     View Summary

    The effects of chronic administration of methamphetamine (MAP) on rat locomotor activity rhythm under light-dark (LD) housing and on neuronal activity rhythms from the suprachiasmatic nucleus (SCN) in vitro were investigated. Control rats exhibited an LD-entrained nocturnal locomotor rhythm. The firing rate of SCN neuronal activity was high during the light period and low during the dark period in control normal rats, and peak of firing activity occurred around zeitgeber time (ZT) 6. On the other hand, chronic MAP administration caused various disorganization of locomotor activity rhythms with a long free-running period (25-35 h). Neuronal activity rhythms of the SCN were unaffected by chronic MAP administration, that is, high during the light period and low during the dark period. The present findings suggested that the SCN maintained as a circadian pacemaker even under chronic MAP administration which affected overt rhythms.

    DOI

  • Effect of the noncompetitive N-methyl-D-aspartate (NMDA) receptor antagonist MK-801 on food-anticipatory activity rhythm in the rat

    M Ono, S Shibata, Y Minamoto, S Watanabe

    PHYSIOLOGY & BEHAVIOR   59 ( 4-5 ) 585 - 589  1996.04

     View Summary

    When feeding was restricted to a single meal scheduled at a fixed time (1300-1700 h) in the day, rats exhibited increased prefeeding locomotor activity 1-3 h before the feeding time, which increased over 6 days. The activity increase was seen for about 5 h(1100-1600 h) on the seventh day when no food was provided (fasting day), as the rat anticipated mealtime. Thus, mealtime-associated activity rhythm on the fasting day was considered to be another good index of food-anticipatory activity. In this study, we investigated the effects of the noncompetitive N-methyl-D-aspartate (NMDA) receptor antagonist, MK-801, which has been reported to cause impairment of spatial learning, on food-anticipatory activity in rats. Daily injection of MK-801 (0.2 mg/kg) at 1700 h for 6 successive days impaired the development of increased prefeeding activity. The appearance of mealtime-associated activity was impaired by daily injection of MK-801 (0.2 mg/kg) at 1700 h (immediately after food restriction, FR) or at 1900 h (2 h after FR) but not at 0900 (16 h after FR). In addition, mealtime-associated activity was impaired by MK-801 injected at 1700 h in a dose-dependent manner(0.1-1 mg/kg). The present results demonstrate that NMDA receptor mechanisms were involved in learning processes of food anticipation and suggest that an impairment of food-anticipatory activity may be considered an animal model of a deficit of ''temporal learning.''

    DOI

  • 体内時計を早めたり遅くしたりする薬

    柴田重信

    現代化学     38 - 44  1996.03

  • 光は替わる・・生体リズムと光

    柴田重信, 守屋孝洋

    照明学会誌   80   28 - 32  1996

    DOI

  • 新規抗パーキンソン氏病薬Talipexoleの鎮静作用に関する脳波学的検討

    河野康子, 柴田重信, 大野益男, 渡辺繁紀

    応用薬理   51   45 - 52  1996

  • Nebracetam (WEB 1881 FU) - A review of its cytoprotective and cholinomimetic properties

    Yasuko Kohno, Shigenobu Shibata

    CNS Drug Reviews   2 ( 1 ) 1 - 20  1996

    DOI

  • Positron-labeled antioxidant 6-deoxy-6-[18F]fluoro-L-ascorbic acid: Increased uptake in transient global ischemic rat brain

    Fumihiko Yamamoto, Shigenobu Shibata, Shigenori Watanabe, Kouji Masuda, Minoru Maeda

    Nuclear Medicine and Biology   23 ( 4 ) 479 - 486  1996  [Refereed]

     View Summary

    The in vivo uptake and distribution of 6-deoxy-6-[18F]fluoro-L-ascorbic acid (18F-DFA) were investigated in rat brains following postischemic reperfusion. Global cerebral ischemia was induced in male Wistar rats for 20 min by occlusion of four major arteries. Two time paints were chosen for 18F-DFA injection to rats subjected to cerebral ischemia, at the start of recirculation and 5 days following recirculation. The rats were then killed at 2 h after tail-vein administration of 18F-DFA and tissue radioactivity concentration was determined. Increased uptake of radioactivity in particular brain regions, including the cerebral cortex, hypothalamus, and amygdala following injection of 18F-DFA, compared to the sham operated control, was observed 5 days after reperfusion. Similar results were also obtained in in vitro experiments using brain slices. Abnormal in vivo accumulation of 45Ca, a marker of regional postischemic injury, was observed in these brain regions in tissue dissection experiments. Furthermore, metabolite analysis of nonradioactive DFA using 19F-NMR showed that DFA remained intact in the postischemic reperfusion brain. The present results indicate that 18F-DFA increasingly accumulates in damaged regions of postischemic reperfusion brain.

    DOI PubMed

  • Calcium channel blockers improve hypoxia/hypoglycemia-induced impairment of rat hippocampal 2-deoxyglucose uptake in vitro after ethanol withdrawal.

    S Watanabe, T Sindou, S Shibata

    NEUROSCIENCES-JAPAN   21 ( 2 ) P177 - P180  1995.12

     View Summary

    The aim of the present study was to determine whether calcium channel antagonists attenuated hypoxia/hypoglycemia- or glutamate-induced reduction in 2-deoxyglucose (2-DG) uptake of hippocampal slices obtained from ethanol withdrawal rats, Ethanol withdrawal significantly potentiated the hypoxia/hypoglycemia- and glutamate-induced reductions in 2-DG uptake of hippocampal slices, Both nifedipine and flunarizine exhibited attenuating effects on ethanol withdrawal-induced potentiation of impairment of 2-DG uptake caused by hypoxia/hypoglycemia or glutamate, Hypoxia/hypoglycemia-induced deficit of 2-DG uptake was prevented by ethanol, but chronic consumption of ethanol resulted in the development of tolerance to neuroprotective effect, These findings suggested that the increased sensitivity of neurons to ischemic damage by ischemia may involve in the increased activity of calcium channels in the hippocampus.

    DOI

  • CIRCADIAN CHANGES IN LONG-TERM POTENTIATION OF RAT SUPRACHIASMATIC FIELD POTENTIALS ELICITED BY OPTIC-NERVE STIMULATION IN-VITRO

    Y NISHIKAWA, S SHIBATA, S WATANABE

    BRAIN RESEARCH   695 ( 2 ) 158 - 162  1995.10

     View Summary

    Optic nerve stimulation caused a postsynaptic field potential in the rat suprachiasmatic nucleus (SCN) of hypothalamic slices. In the present experiment, we demonstrated whether tetanic stimulation of optic nerve can produce a long-term potentiation (LTP) in the SCN postsynaptic field potential. The amplitude of SCN field potential was higher in the subjective day animals than that in the subjective night animals. Tetanic stimulation of optic nerve (100 Hz, 1 s) at subjective daytime (projected zeitgeber time: ZT 0-8) produced a LTP in this field potential, although the onset of LTP was slow. When tetanic stimulation was applied at ZT4, the percent increase of amplitude was 116.6% immediately after, 159.8% 30 min after and 215.4% 120 min after tetanic stimulation, whereas tetanic stimulation of optic nerve at subjective night-time caused a weak LTP in the SCN. Although tetanic stimulation of Schaffer collaterals induced a LTP formation in the CA1 region of rat hippocampal slices, there were no obvious circadian changes in this LTP formation. The present results demonstrated that excitatory influence on the SCN caused a synaptic plasticity such as LTP. Although the physiological meaning of this LTP is uncertain at present, LTP may be related to adaptation mechanism to photic stimulation.

    DOI

  • CIRCADIAN-RHYTHM OF SPONTANEOUS NEURONAL-ACTIVITY IN THE SUPRACHIASMATIC NUCLEUS OF OLD HAMSTER IN-VITRO

    A WATANABE, S SHIBATA, S WATANABE

    BRAIN RESEARCH   695 ( 2 ) 237 - 239  1995.10

     View Summary

    The effects of aging on neuronal activity in the suprachiasmatic nucleus (SCN) were examined in hamsters kept under light-dark (LD) or constant light (LL) conditions. The free-running period in wheel-running rhythm of 24-month-old hamsters (24.2 +/- 0.04) was shorter than that of the 2-month-old hamsters (24.4 +/- 0.057). There was a significant difference in the mean firing rates of SCN neuron activity between old and young hamsters during subjective day (6.58 +/- 0.36 spikes/s in young and 5.63 +/- 0.24 in old hamsters), but not during subjective night (4.33 +/- 0.47 in young and 4.05 +/- 0.39 in old). Similar to LL condition, the firing activity during zeitgeber time 3-11 (4.33 +/- 0.27) in old hamsters kept under LD condition, was significantly lower than that of young hamsters (6.22 +/- 0.32). These results suggest that deterioration of SCN neuronal activity in old hamsters assessed as reduction of daytime activity may reflect changes in the interaction between SCN clocks and the overt behaviors and/or pacemaking properties of SCN cells.

    DOI

  • METHYLCOBALAMIN ATTENUATES THE HYPOXIA/HYPOGLYCEMIA-INDUCED OR GLUTAMATE-INDUCED REDUCTION IN HIPPOCAMPAL FIBER SPIKES IN-VITRO

    Y YAMAMOTO, S SHIBATA, C HARA, S WATANABE

    EUROPEAN JOURNAL OF PHARMACOLOGY   281 ( 3 ) 335 - 340  1995.08

     View Summary

    The effects of methylcobalamin, a vitamin B-12 analogue, on the hypoxia/hypoglycemia- or glutamate-induced reduction in hippocampal CA1 presynaptic fiber spikes elicited by Schaffer collateral stimulation in rat brain slices were evaluated. Hippocampal slices were exposed to 15 min of hypoxia/hypoglycemia, and then these slices were returned to oxygenated and glucose-containing buffer for 3 h. Hypoxia/hypoglycemia reduced CA1 presynaptic potentials in vitro. Treatment with 10 mu M methylcobalamin attenuated the impairment of CA1 presynaptic potentials induced by hypoxia/hypoglycemia or glutamate application (10 mM). Daily injection of methylcobalamin (0.5 mg/kg i.p./day) for 3 days in vivo also attenuated the hypoxia/hypoglycemia- or glutamate-induced reduction in presynaptic potentials in hippocampal slices. Pretreatment with cyanocobalamin at 10 mu M failed to attenuate the impairment of CA1 presynaptic potentials. However, daily injection of cyanocobalamin (0.5 mg/kg i.p./day) for 3 days caused a protective action against the hypoxia/hypoglycemia- or glutamate-induced functional deficit. Furthermore, co-treatment of L-arginine (100 mu M), a substrate for nitric oxide synthase, with methylcobalamin in vitro reversed the methylcobalamin-induced functional recovery. The present results demonstrate that methylcobalamin application in vivo or in vitro leads to functional recovery from hypoxia/hypoglycemia- or glutamate-induced impairment of CA1 presynaptic potentials. Neuroprotection was obtained by in vivo application of cyanocobalamin, but not by its in vitro application. It is reported that in vivo injected cyanocobalamin converted to methylcobalamin in the hepatic cells. Therefore, the results suggest that a transmethylation reaction in the hippocampal regions may be involved in the methylcobalamin-induced functional recovery from ischemic impairment.

    DOI

  • INCREASE IN THE SEPTAL VASOPRESSIN CONTENT BY PROLYL ENDOPEPTIDASE INHIBITORS IN RATS

    N MIURA, S SHIBATA, S WATANABE

    NEUROSCIENCE LETTERS   196 ( 1-2 ) 128 - 130  1995.08

     View Summary

    Prolyl endopeptidase (PEP; EC 3.4.21.26) cleaves the Pro-Arg bond of arginine-vasopressin (AVP), This study investigated the effects of PEP inhibitors, 1-[1-(benzyloxycarbonyl)-L-prolyl]prolinal (Z-Pro-prolinal) and 1-[3-(2-indanylacetyl)-L-thioprolyl]pyrrolidine (Z-321), on the AVP content in the septum of rats. Oral administration of Z-Pro-prolinal (100 mg/kg) and Z-321 (100 mg/kg) significantly increased the septal AVP content. At 10 mg/kg, Z-321 slightly increased the content. In contrast, the D-thioprolyl form of Z-321 (100 mg/kg), which lacks an inhibitory effect on the enzyme, failed to affect the AVP content. These results indicate that PEP inhibitors increase the content of AVP through inhibition of the enzyme activity in vivo. Therefore, it is suggested that PEP may contribute to the degradation of endogenous AVP in the brain.

    DOI

  • INVOLVEMENT OF DOPAMINE, N-METHYL-D-ASPARTATE AND SIGMA-RECEPTOR MECHANISMS IN METHAMPHETAMINE-INDUCED ANTICIPATORY ACTIVITY RHYTHM IN RATS

    S SHIBATA, M ONO, N FUKUHARA, S WATANABE

    JOURNAL OF PHARMACOLOGY AND EXPERIMENTAL THERAPEUTICS   274 ( 2 ) 688 - 694  1995.08

     View Summary

    Daily injection of methamphetamine (MAP) increased the locomotor activity and produced the development of MAP-induced anticipatory activity on the next withdrawal day. Daily restricted feeding also causes a feeding-associated anticipatory activity rhythm. The importance of catecholaminergic neurons has been suggested in the manifestation of a feeding-associated corticosterone rhythm. In our study, we examined the role of dopaminergic neurons in the development of MAP-induced anticipatory activity. The development of MAP-induced anticipatory activity was blocked by coadministration of dopamine D2 receptor antagonists such as YM-09151-2 and sulpiride, the D1 receptor antagonist SCH23390 acid haloperidol weak D1/D2 receptor antagonist, but not by clozapine. The anticipatory activity increase was reproduced by daily injection of the D2 receptor agonist quinpirole as well as moderately by the D1 receptor agonist SKF38393. Moreover, MAP-induced anticipation was blocked by coadministration of the N-methyl-D-aspartate receptor antagonist MK-801 or sigma ligands rimcazol and NE-100. Our results suggest that activation of the dopaminergic system, especially via D2 receptors may be required for the development of MAP-associated anticipatory activity increases and that N-methyl-D-aspartate receptors and sigma receptor mechanisms are also involved in the development of this behavior.

  • κ-Opioid receptor agonist protects against ischemic reduction of 2-deoxyglucose uptake in morphine-tolerant rats

    Shigenobu Shibata, Keiko Tominaga, Shigenori Watanabe

    European Journal of Pharmacology   279 ( 2-3 ) 197 - 202  1995.06  [Refereed]

     View Summary

    We examined the effects of μ-opioid receptor agonist and antagonists, and κ-opioid receptor agonist on the hypoxia/hypoglycemia-induced reduction in 2-deoxyglucose uptake of rat hippocampal slices. Naloxone, a μ-opioid receptor antagonist and (5,7,8)-(+)-3,4-dichloro-N-methyl-N-(7,8,1-pyrrolidinyl)-1-oxaspirol 4,5 dec-8-yl)-benzeneacetamide methanesulfonate, U-62,066E, a κ-opioid receptor agonist, showed neuroprotective actions against the hypoxia/hypoglycemia-induced deficit in glucose uptake. In contrast, morphine exhibited an exacerbating action. These results suggest that blockade of μ-opioid receptor-and stimulation of κ-opioid receptor-mediated functions has a protective role against the hypoxia/hypoglycemia-induced decreases in glucose metabolism in hippocampal slices. Chronic administration of morphine (10 mg/kg) for 9 days affected neither the basal nor the hypoxia/hypoglycemia-induced reduction in 2-deoxyglucose uptake. Rats treated with morphine chronically exhibited not only tolerance to the analgesic effect but also tolerance to the exacerbating action. However, chronic morphine did not modify U-62,066E-induced neuroprotection. These findings indicate that the receptor mechanisms of neuroprotection produced by the activation of κ-opioid receptors may not be involved in μ-opioid receptor function. © 1995.

    DOI PubMed

  • EFFECT OF A NITRIC-OXIDE SYNTHASE INHIBITOR, N-NITRO-L-ARGININE METHYLESTER, ON LIGHT-INDUCED PHASE DELAY OF CIRCADIAN-RHYTHM OF WHEEL-RUNNING ACTIVITY IN GOLDEN-HAMSTERS

    A WATANABE, M ONO, S SHIBATA, S WATANABE

    NEUROSCIENCE LETTERS   192 ( 1 ) 25 - 28  1995.06

     View Summary

    Under constant darkness hamsters demonstrate free-running activity rhythms and light exposure during the early subjective night results in permanent phase delays of the activity rhythm. Recently, we reported that application of glutamate receptor agonists such as N-methyl-D-aspartate could reset the phase of the circadian rhythm of suprachiasmatic nucleus firing activity in vitro via nitric oxide production. In order to confirm this result by in vivo experiment, we examined the effect of nitric oxide synthesis inhibitor on the light-induced phase delay of circadian rhythms of wheel-running activity in hamsters. In vehicle-treated animals, light stimulation at circadian time 13.5 resulted in stable phase delays (1.3 +/- 0.63 h), whereas pre-treatment with 150 mg/kg of N-nitro-L-arginine methylester (L-NAME) significantly attenuated light-induced phase delays (0.72 +/- 0.18 h). L-NAME administration alone without light exposure, did not cause phase changes. The L-NAME-induced attenuating effect was reversed by co-administration of L-arginine (300 mg/kg). The present results suggest that nitric oxide production is involved in the light-induced phase delay of the hamster's circadian system.

    DOI

  • ATTENUATING EFFECT OF SEROTONIN RECEPTOR ANTAGONISTS ON IMPAIRMENT OF MEALTIME-ASSOCIATED ACTIVITY RHYTHM IN OLD RATS

    S SHIBATA, M ONO, Y MINAMOTO, S WATANABE

    PHARMACOLOGY BIOCHEMISTRY AND BEHAVIOR   51 ( 2-3 ) 541 - 544  1995.06

     View Summary

    In the present study, we examined attenuating effect of serotonin (5-HT) receptor antagonists on the impairment of the time perception presented by daily scheduled feeding in old rats. When feeding was restricted to a single meal at a fixed time of day (1300-1700 h) for six consecutive days, young rats exhibited intense locomotor activity from 1-3 h before feeding time. Intense locomotor activity was observed between 1200 and 1700 h in young animals even on the fasting day (day 7) (mealtime-associated activity). However, this mealtime-associated activity was impaired in 24-mo-old rats. Daily injections of 5-HT2 receptor antagonists, mianserin or ritanserin, or a 5-HT3 receptor antagonist, Y25130, at 1700 h for 6 consecutive days significantly and dose-dependently attenuated the impairment of mealtime-associated activity on the fasting day in old rats without affecting the food intake. Our results suggest that the blockade of 5-HT2 and/or 5-HT3 receptors attenuates impairment of the manifestation of mealtime-associated anticipatory activity related to temporal learning in old rats.

    DOI

  • LONG-TERM ENHANCEMENT OF DOPAMINE RELEASE BY HIGH-FREQUENCY TETANIC STIMULATION VIA A N-METHYL-D-ASPARTATE-RECEPTOR-MEDIATED PATHWAY IN RAT STRIATUM

    M OCHI, H INOUE, S KOIZUMI, S SHIBATA, S WATANABE

    NEUROSCIENCE   66 ( 1 ) 29 - 36  1995.05

     View Summary

    We studied the effects of high frequency tetanic stimulation of the striatum on the KCl (20 mM)-evoked dopamine release in rat striatal slices. The KCl-evoked dopamine release was potentiated by high frequency tetanic stimulation (10-20 Hz) of the striatum including the corticostriatal fibers, and this potentiation was observed until 3 h after high frequency tetanic stimulation. Potentiation of dopamine release after high frequency tetanic stimulation was induced not only by KCl but also by glutamate in Mg2+-free medium, N-methyl-D-aspartate in Mg2+-free medium, and by DL-alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid. 2-amino-5-phosphovalerate, 3-[(+/-)-2-carboxypiperazine-4-yl]-propy-1-phosphonate or dibenzocycloheptaneimine, N-methyl-D-aspartate receptor inhibitors, abolished enhancement by tetanus, whereas, 6,7-dinitroquinoxaline-2,3-dione, an antagonist of DL-alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid ionotropic receptors, or L-2-amino-4-phosphonobutyrate, an antagonist of glutamate metabotropic receptors, showed no effect. Moreover, pretreatment with glutamate or N-methyl-D-aspartate in the absence of Mg2+ also facilitated dopamine release evoked by KCl concentrations. When extracellular Ca2+ was removed from the medium during pretreatment, potentiation by glutamate disappeared.
    We conclude that activation of N-methyl-D-aspartate receptors on dopaminergic nerve terminals in the striatum produces the long-term changes in efficacy of the response to KCl or glutamatergic agents. That is, plastical phenomena could exist at presynaptic levels between glutamatergic neurons and dopaminergic neurons in striatum.

    DOI

  • Calcium channel blockers improve hypoxia/hypoglycemia-induced impairment of rat hippocampal 2-deoxyglucose uptake in vitro after ethanol withdrawal

    Shigenobu Shibata, Tomomi Shindou, Keiko Tominaga, Shigenori Watanabe

    Brain Research   673 ( 2 ) 320 - 324  1995.03  [Refereed]

     View Summary

    The aim of the present study was to determine whether calcium channel antagonists attenuated hypoxia/hypoglycemia- or glutamate-induced reduction in 2-deoxyglucose (2-DG) uptake of hippocampal slices obtained from ethanol withdrawal rats. Ethanol withdrawal significantly potentiated the hypoxia/hypoglycemia- and glutamate-induced reductions in 2-DG uptake of hippocampal slices. Both nifedipine and flunarizine exhibited attenuating effects on ethanol withdrawal-induced potentiation of impairment of 2-DG uptake caused by hypoxia/hypoglycemia or glutamate. Hypoxia/hypoglycemia-induced deficit of 2-DG uptake was prevented by ethanol, but chronic consumption of ethanol resulted in the development of tolerance to neuroprotective effect. These findings suggest that the increased sensitivity of neurons to ischemic damage by ischemia may involve in the increased activity of calcium channels in the hippocampus. © 1995 Elsevier Science B.V. All rights reserved.

    DOI PubMed

  • EFFECTS OF TRANSIENT FOREBRAIN ISCHEMIA ON LONG-TERM ENHANCEMENT OF DOPAMINE RELEASE IN RAT STRIATAL SLICES

    H INOUE, M OCHI, S SHIBATA, S WATANABE

    BRAIN RESEARCH   671 ( 1 ) 95 - 99  1995.02

     View Summary

    We studied the effects of transient forebrain ischemia in vivo on long-term enhancement of dopamine (DA) release from rat striatal slices. One hour after the high-frequency tetanic stimulation (HFTS) or L-glutamate (10(-6) M) application in Mg2+-free medium to striatal slices, the high concentration of KCl (high K+)-evoked DA release was measured. Tetanic stimulation or L-glutamate application significantly potentiated the high-K+-evoked DA release. When striatal slices were prepared from rats exposed to 3 min of ischemia followed by 24-h survival, the enhancement of DA release by HFTS was unaffected by ischemia. In contrast, the enhancement of DA release by HFTS was impaired in rats exposed to 5 min or 10 min of ischemia. In addition, high K+-evoked DA release per se was significantly impaired by 10 min of ischemia. The enhancement of DA release elicited by pretreatment with L-glutamate was also impaired in the rats exposed to 5 min of ischemia. When striatal slices were prepared from rats exposed to 5 min of ischemia with 7-day survival, the enhancement of DA release by HFTS was still impaired. The present results indicate that the neuronal mechanisms of the enhancement of DA release may be more sensitive to impairement from short periods of ischemia. Furthermore, the results suggest that an impairment of long-term enhancement of DA release by ischemia may be related the dysfunction of motor performance in rats exposed to ischemia.

    DOI

  • N-G-NITRO-L-ARGININE PROTECTS AGAINST HYPOXIA HYPOGLYCEMIA-INDUCED DECREASE IN CA1 PRESYNAPTIC SPIKES IN RAT HIPPOCAMPAL SLICES

    S SHIBATA, Y YAMAMOTO, T TANAKA, S WATANABE

    EUROPEAN JOURNAL OF PHARMACOLOGY   273 ( 3 ) 223 - 228  1995.02

     View Summary

    The effects of nitric oxide (NO) synthase inhibitors on the hypoxia/hypoglycemia-induced decrease in CA1 presynaptic fiber spikes elicited by stimulation of the Schaffer collaterals were investigated using rat hippocampal slices. Drugs were added to normal medium for 10 min before incubation under hypoxic/hypoglycemic conditions (15 min), and after a 3-h washout, the CA1 presynaptic potential was measured. Treatment with N-G-nitro-L-arginine methyl ester but not with N-G-nitro-D-arginine methyl ester produced a concentration-dependent attenuation of the hypoxia/hypoglycemia-induced decrease in presynaptic fiber spikes. In contrast, treatment with precursors of NO in the arginine-to-NO pathway, such as sodium nitroprusside, S-nitro-N-acetylpenicillamine and N-morpholino sydnonimine exacerbated the 15-min hypoxia/hypoglycemia-induced decrease in the CA1 presynaptic potential. The neuroprotective effect of N-G-nitro-L-arginine methyl aster was significantly attenuated by co-treatment with L-arginine. The present results suggest a facilitatory role of NO production in hypoxia/hypoglycemia-induced presynaptic dysfunction in CA1 regions of hippocampal slices.

    DOI

  • N-G-NITRO-L-ARGININE PROTECTS AGAINST HYPOXIA HYPOGLYCEMIA-INDUCED DECREASE IN CA1 PRESYNAPTIC SPIKES IN RAT HIPPOCAMPAL SLICES

    S SHIBATA, Y YAMAMOTO, T TANAKA, S WATANABE

    EUROPEAN JOURNAL OF PHARMACOLOGY   273 ( 3 ) 223 - 228  1995.02

     View Summary

    The effects of nitric oxide (NO) synthase inhibitors on the hypoxia/hypoglycemia-induced decrease in CA1 presynaptic fiber spikes elicited by stimulation of the Schaffer collaterals were investigated using rat hippocampal slices. Drugs were added to normal medium for 10 min before incubation under hypoxic/hypoglycemic conditions (15 min), and after a 3-h washout, the CA1 presynaptic potential was measured. Treatment with N-G-nitro-L-arginine methyl ester but not with N-G-nitro-D-arginine methyl ester produced a concentration-dependent attenuation of the hypoxia/hypoglycemia-induced decrease in presynaptic fiber spikes. In contrast, treatment with precursors of NO in the arginine-to-NO pathway, such as sodium nitroprusside, S-nitro-N-acetylpenicillamine and N-morpholino sydnonimine exacerbated the 15-min hypoxia/hypoglycemia-induced decrease in the CA1 presynaptic potential. The neuroprotective effect of N-G-nitro-L-arginine methyl aster was significantly attenuated by co-treatment with L-arginine. The present results suggest a facilitatory role of NO production in hypoxia/hypoglycemia-induced presynaptic dysfunction in CA1 regions of hippocampal slices.

    DOI

  • ATTENUATING EFFECT OF BIFEMELANE ON AN IMPAIRMENT OF MEALTIME-ASSOCIATED ACTIVITY RHYTHM IN AGED AND MK-801-TREATED RATS

    S SHIBATA, M ONO, Y MINAMOTO, S WATANABE

    PHARMACOLOGY BIOCHEMISTRY AND BEHAVIOR   50 ( 2 ) 207 - 210  1995.02

     View Summary

    In the present experiment, we examined the attenuating effect of bifemelane hydrochloride (BF), 4-(o-benzyl phenoxy)-N-methylbutylamine hydrochloride, on the impairment of time perception caused by daily scheduled feeding using aged and MK-801-treated rats. When feeding was restricted to a single meal at a fixed time of day (1300-1700 h) for six successive days, young rats exhibited intense locomotor activity 1-3 h before feeding time. Intense locomotor activity was observed for 1200-1700 h even on the fasting day (day 7; mealtime-associated activity). Mealtime-associated activity was impaired in 24-mo-old rats and also in N-methyl-D-aspartate receptor antagonist, MK-801-treated rats. Daily injections of bifemelane at 1700 h for six successive days significantly attenuated the impairment of mealtime-associated activity on the seventh day in a dose-dependent manner in aged rats. In addition, cotreatment of MK-801 with bifemelane blocked the MK-801-induced impairment of mealtime-associated activity. The present study suggests that bifemelane has an enhancing effect on learning and memory performance, such as spatial and temporal perception.

    DOI

  • Long-term enhancement of dopamine release by high frequency tetanic stimulation via aN-methyl-d-aspartate-receptor-mediated pathway in rat striatum

    M. Ochi, H. Inoue, S. Koizumi, S. Shibata, S. Watanabe

    Neuroscience   66 ( 1 ) 29 - 36  1995

     View Summary

    We studied the effects of high frequency tetanic stimulation of the striatum on the KCl (20 mM)-evoked dopamine release in rat striatal slices. The KCl-evoked dopamine release was potentiated by high frequency tetanic stimulation (10-20 Hz) of the striatum including the corticostriatal fibers, and this potentiation was observed until 3 h after high frequency tetanic stimulation. Potentiation of dopamine release after high frequency tetanic stimulation was induced not only by KCl but also by glutamate in Mg2+-free medium,N-methyl-d-aspartate in Mg2+-free medium, and bydl-α-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid. 2-amino-5-phosphovalerate, 3-[(±)-2-carboxypiperazine-4-yl]-propyl-1-phosphonate or dibenzocycloheptaneimine,N-methyl-d-aspartate receptor inhibitors, abolished enhancement by tetanus, whereas, 6,7-dinitroquinoxaline-2,3-dione, an antagonist ofdl-α-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid ionotropic receptors, orl-2-amino-4-phosphonobutyrate, an antagonist of glutamate metabotropic receptors, showed no effect. Moreover, pretreatment with glutamate orN-methyl-d-aspartate in the absence of Mg2+ also facilitated dopamine release evoked by KCl concentrations. When extracellular Ca2+ was removed from the medium during pretreatment, potentiation by glutamate disappeared. We conclude that activation ofN-methyl-d-aspartate receptors on dopaminergic nerve terminals in the striatum produces the long-term changes in efficacy of the response to KCl or glutamatergic agents. That is, plastical phenomena could exist at presynaptic levels between glutamatergic neurons and dopaminergic neurons in striatum. © 1995 IBRO.

    DOI PubMed

  • 光による体内時計リセットの情報伝達機構

    柴田重信

    蛋白質核酸酵素   40 ( 16 ) 2408 - 2417  1995

  • ATTENUATING EFFECT OF ARECOLINE AND PHYSOSTIGMINE ON AN IMPAIRMENT OF MEALTIME-ASSOCIATED ACTIVITY RHYTHM IN OLD RATS

    M ONO, Y MINAMOTO, S SHIBATA, S WATANABE

    PHYSIOLOGY & BEHAVIOR   57 ( 1 ) 189 - 191  1995.01

     View Summary

    In the present study, we examined whether cholinergic drugs such as arecoline and physostigmine attenuated an impairment of time perception presented by daily scheduled feeding in aged rats. When feeding was restricted to a single meal at a fixed time of day (13:00-17:00) for 6 successive days, young rats exhibited intense locomotor activity from 1-3 h before feeding time. Intense locomotor activity was observed between 12:00-17:00 in young animals even on the fasting day (on day 7) (mealtime-associated activity). However, this mealtime-associated activity was impaired in old rats. Daily injection of arecoline (10 mg/kg) or physostigmine (0.1 and 0.2 mg/ kg) at 17:00 for 6 successive days attenuated the impairment of mealtime associated activity on the fasting day in a dose-dependent manner in old rats, whereas daily treatment with D-glucose (100 or 2000 mg/kg) did not. The results of the present study suggest that cholinergic drugs attenuate the impairment of the manifestation of mealtime-associated anticipatory activity related to 'temporal learning' in old rats.

    DOI

  • GLUTATHIONE PROTECTS AGAINST HYPOXIC HYPOGLYCEMIC DECREASES IN 2-DEOXYGLUCOSE UPTAKE AND PRESYNAPTIC SPIKES IN HIPPOCAMPAL SLICES

    S SHIBATA, K TOMINAGA, S WATANABE

    EUROPEAN JOURNAL OF PHARMACOLOGY   273 ( 1-2 ) 191 - 195  1995.01

     View Summary

    The effects of glutathione, its analogue: YM737 (N-(N-gamma-L-glutamyl-L-cysteinyl) glycine-l-isopropyl ester sulfate monohydrate), a monoester of glutathione, and N-acetyl-L-cysteine on hypoxia/hypoglycemia-induced decreases in CA1 presynaptic fiber spikes and 2-deoxyglucose uptake were investigated using rat hippocampal slices. The drugs were added to normal medium for 30 min before the incubation under hypoxic/hypoglycemic conditions (20 min), and, after a 3-h washout, presynaptic potential or 2-deoxyglucose uptake in hippocampal slices was measured. Treatment with glutathione, YM737 and N-acetyl-L-cysteine produced an attenuation of the hypoxia/hypoglycemia-induced decrease in presynaptic fiber spikes and 2-deoxyglucose uptake. The order of potency for neuroprotective action was YM737 greater than or equal to N-acetyl-L-cysteine &gt; glutathione. The present results suggest a role for glutathione in improving hypoxia/hypoglycemia-induced dysfunction of hippocampal regions.

    DOI

  • A ROLE OF SIGMA-RECEPTORS ON HYPOXIA HYPOGLYCEMIA-INDUCED DECREASE IN CA1 PRESYNAPTIC FIBER SPIKES IN RAT HIPPOCAMPAL SLICES

    S SHIBATA, Y YAMAMOTO, S WATANABE

    BRAIN RESEARCH   670 ( 2 ) 337 - 341  1995.01

     View Summary

    Effects of sigma receptor agonists or antagonists on hypoxia/hypoglycemia-induced decrease in CA1 presynaptic fiber spikes elicited by the stimulation of Schaffer collaterals were investigated using rat hippocampal slices. Treatment with sigma receptor antagonists such as haloperidol, NE-100 and rimcazole produced a concentration-dependent attenuation of the hypoxia/hypoglycemia-induced decrease in CA1 presynaptic fiber spikes. The order of potency of protection against hypoxia/ hypoglycemia-induced reduction in CA1 presynaptic potential was: NE-100 = haloperidol &gt; rimcazole. Treatment with sigma receptor agonist DTG potentiated the hypoxia/hypoglycemia-induced decrease in the CA1 presynaptic potential, whereas SKF10047 which possesses an affinity for phencyclidine site attenuated the decrease of potential. NE-100 antagonized a functional deficit induced by DTG, but unaffected the improving effect induced by SKF10047. The present results suggest a facilitatory role of sigma receptor stimulation in hypoxia/hypoglycemia-induced an impairment of neurophysiological functions in CA1 presynaptic regions of hippocampal slices.

    DOI

  • Effects of naloxone, morphine and κ-opioid receptor agonists on hypoxia/hypoglycemia-induced reduction of 2-deoxyglucose uptake in hippocampal slices from U-50,488H-tolerant rats

    Shigenobu Shibata, Keiko Tominaga, Shigenori Watanabe

    Neuroscience Letters   182 ( 2 ) 155 - 158  1994.12  [Refereed]

     View Summary

    The aim of the present study was to determine whether U-50,488H and U-62,066E, κ-opioid receptor agonists cause a neuroprotective action against hypoxia/hypoglycemia-induced reduction in 2-deoxyglucose (2-DG) uptake of hippocampal slices from U-50,488H-tolerant rats. Both U-50,488H and U-62,066E exhibited an attenuating effect on hypoxia/hypoglycemia-induced reduction in 2-DG uptake of hippocampal slices. Hypoxia/hypoglycemia-induced deficit of 2-DG uptake was prevented by cotreatment with naloxone, an opioid receptor antagonist, but potentiated by cotreatment with morphine, a μ-opioid receptor agonist. Chronic administration of U-50,488H resulted in the development of tolerance to the analgesic effect as well as the neuroprotective effect whereas this treatment affected neither basal- nor hypoxia/hypoglycemia-induced decreases in 2-DG uptake. Chronic administration of U-50,488H did not modify naloxone-induced attenuation of 2-DG uptake deficit but slightly potentiated the morphine-induced exacerbation. These findings suggest that the tolerance to κ-opioid receptors does not affect the μ-opioid receptor-mediated neuroprotective or neurotoxic action. © 1994.

    DOI PubMed

  • PHASE ADVANCES OF CIRCADIAN-RHYTHMS IN SOMATOSTATIN DEPLETED RATS - EFFECTS OF CYSTEAMINE ON RHYTHMS OF LOCOMOTOR-ACTIVITY AND ELECTRICAL-DISCHARGE OF THE SUPRACHIASMATIC NUCLEUS

    C FUKUHARA, T HAMADA, S SHIBATA, S WATANABE, K AOKI, SIT INOUYE

    JOURNAL OF COMPARATIVE PHYSIOLOGY A-SENSORY NEURAL AND BEHAVIORAL PHYSIOLOGY   175 ( 6 ) 677 - 685  1994.12

     View Summary

    Somatostatin is synthesized in the suprachiasmatic nucleus (SCN), a circadian pacemaker in mammals. To explore the functional significance of somatostatin in the circadian system, we examined rhythms of rat locomotor activity and electrical firing rate of SCN neurons in the brain slice after temporal depletion of somatostatin levels in the SCN. Intraperitoneal administration of cysteamine (200 mg/kg), a somatostatin depletor, significantly reduced somatostatin level in the in vivo SCN 5 min after injection and kept low level as long as 3 to 4 days. This administration, on the other hand, induced significant phase advances of about 51 min in the subsequent free-running rhythm of locomotor activity of the rat. A marked phase advance in the circadian rhythm of firing rate in the SCN was also observed after administration of cysteamine in coronal hypothalamic slices. These persistent phase shifts after administration of a somatostatin depletor may suggest that the change of somatostatin level in the SCN have a feedback influence on the circadian pacemaker.

    DOI

  • FACILITATORY EFFECT OF OLFACTORY BULBECTOMY ON 2-DEOXYGLUCOSE UPTAKE IN RAT AMYGDALA SLICES

    S SHIBATA, S WATANABE

    BRAIN RESEARCH   665 ( 1 ) 147 - 150  1994.11

     View Summary

    The aim of the present study was to determine whether the 2-deoxyglucose uptake of amygdala slices was affected by olfactory bulbectomy. At 7, 14 and 21 days post-lesion, bilateral olfactory bulbectomized rats exhibited a significant increase of 2-deoxyglucose uptake in amygdala slices, but not in the cerebral cortex. In addition, unilateral olfactory bulbectomized rats showed a high uptake of 2-deoxyglucose uptake in the ipsilateral amygdala, but not in the contralateral amygdala. These results suggest that the enhancement of 2-deoxyglucose uptake is related to the hyperexcitability of amygdala neurons following bulbectomy, and that this enhancement may be responsible for the behavioral changes in olfactory bulbectomized rats.

    DOI

  • INVOLVEMENT OF D-1 DOPAMINE-RECEPTOR MECHANISM IN ISCHEMIA-INDUCED IMPAIRMENT OF CA1 PRESYNAPTIC FIBER SPIKES IN RAT HIPPOCAMPAL SLICES

    Y YAMAMOTO, T TANAKA, S SHIBATA, S WATANABE

    BRAIN RESEARCH   665 ( 1 ) 151 - 154  1994.11

     View Summary

    The effect of dopamine (DA) receptor agonists and antagonists on hypoxia/hypoglycemia (ischemia)-induced decrease in CA1 presynaptic fiber spikes elicited by the stimulation of Schaffer collateral were investigated using hippocampal slices. Treatment with D-1 dopamine receptor antagonist, SCH23390 produced a concentration-dependent attenuation of the ischemia-induced decrease of presynaptic potentials. The magnitude of recovery of the CA1 presynaptic potential in SCH233390-treated slices at 10 and 100 mu M was 28 and 54%, respectively. Whereas, treatment with D1 dopamine receptor agonist, SKF38393 exacerbated the ischemia-induced decrease in the CA1 presynaptic potential. The decrease of CA1 presynaptic potential by ischemia was affected by neither D-2 dopamine receptor agonist, bromocriptin and quinpirole nor D-2 dopamine receptor antagonist, sulpiride. The neuroprotective effect of SCH23390 was completely blocked by cotreatment with SKF38393. The present results demonstrated that the blockade of D-1 dopamine receptor function played a neuroprotective role in ischemic damage, suggesting a facilitatory role of D-1 dopamine receptor-operated function in ischemia-induced neuronal deficits.

    DOI

  • FACILITATORY EFFECTS OF SOMATOSTATIN ON REDUCED UPTAKE OF 2-DEOXYGLUCOSE IN CEREBRAL CORTICAL AND HIPPOCAMPAL SLICES FROM AGED RATS

    S SHIBATA, T TANAKA, S WATANABE

    EUROPEAN JOURNAL OF PHARMACOLOGY-MOLECULAR PHARMACOLOGY SECTION   269 ( 2 ) 269 - 272  1994.10

     View Summary

    The aim of the present study was to determine whether or not the reduction of 2-deoxyglucose uptake by the cerebral cortical slices in aged rats (22-23 months old) was attenuated by somatostatin or carbachol. In 8-week-old rats, somatostatin and carbachol produced concentration-dependent increases in 2-deoxyglucose uptake. 2-Deoxyglucose uptake of the cortical slices in 22-23-month-old rats was significantly facilitated by treatment with 0.1-1 mu M somatostatin or 1-100 mu M carbachol. Metabolic responses to somatostatin or carbachol were quite similar in young and aged rats. The present results demonstrated that 2-deoxyglucose uptake by the cerebral cortex was facilitated by somatostatin and carbachol in both young and old rats.

    DOI

  • NEUROCHEMICAL ORGANIZATION OF CIRCADIAN-RHYTHM IN THE SUPRACHIASMATIC NUCLEUS

    SIT INOUYE, S SHIBATA

    NEUROSCIENCE RESEARCH   20 ( 2 ) 109 - 130  1994.08

     View Summary

    The circadian rhythm in mammals is under control of the pacemaker located in the suprachiasmatic nucleus (SCN) of the hypothalamus. This tiny nucleus contains a number of neurochemicals, including peptides, amines and amino acids. Heterogeneous distribution of these neurochemicals defines the substructures of the SCN. In the present review, functional significance of such neurochemical heterogeneity in the SCN is discussed in the light of circadian patterns of the concentrations of these neurochemicals in the SCN and their effects on SCN neurons in in vitro slice preparation. In particular, the hypothesis that the dorsomedial SCN is involved in maintaining the circadian rhythm, while the ventrolateral SCN is involved in adjusting the phase of the rhythm, is critically discussed. These considerations suggest that distinct sub-components of the SCN as marked by neurochemicals, interact with each other and this organizational architecture could be the basis of the proper operation of the circadian time keeping system in this nucleus.

    DOI

  • N-methyl-D-aspartate induces phase shifts in circadian rhythm of neuronal activity of rat SCN in vitro.

    Shibata S, Watanabe A, Hamada T, Ono M, Watanabe S

    The American journal of physiology   267   R360 - 4  1994.08  [Refereed]

    PubMed

  • AGE-RELATED IMPAIRMENT OF FOOD ANTICIPATORY LOCOMOTOR-ACTIVITY IN RATS

    S SHIBATA, Y MINAMOTO, M ONO, S WATANABE

    PHYSIOLOGY & BEHAVIOR   55 ( 5 ) 875 - 878  1994.05

     View Summary

    In rats of various ages (11 weeks to 24 months), restricted feeding was carried for 6 successive days and food was withheld completely on the 7th day to assess the effect of aging on food anticipatory activity. When feeding was restricted to a single meal at a fixed time of day (1330-1730), rats exhibited intense locomotor activity from 1-3 h before feeding time (prefeeding activity). This prefeeding activity was still detected on the fasting day. In addition, intense locomotor activity was observed for 1330-1730 even on the fasting day (mealtime-associated activity). Thus, mealtime-associated activity of the fasting day became an another good index for food anticipatory activity. These results indicate that the rats had come to anticipate the mealtime. Both prefeeding and mealtime-associated activities were similar at 11 weeks and 9 months of age, but they were impaired at 12 and 24 months of age. In 11-week-old rats, there was a negative correlation between the level of mealtime-associated activity and the daily duration of access to food, but this relationship was absent in 12-month-old rats. The present study suggests that aging strongly impairs the manifestation of food anticipatory activity in rats.

    DOI

  • FACILITATORY EFFECT OF VASOPRESSIN ON THE ISCHEMIC DECREASE OF THE CA1 PRESYNAPTIC FIBER SPIKES IN RAT HIPPOCAMPAL SLICES

    T TANAKA, Y SHISHIDO, S SHIBATA, S WATANABE

    BRAIN RESEARCH   644 ( 2 ) 343 - 346  1994.05

     View Summary

    We investigated the effects of vasopressin-related neuropeptides on the hypoxia/hypoglycemia (ischemia)-induced decrease of the CAI presynaptic potential elicited by stimulation of Schaffer collaterals in rat hippocampal dices. Treatment with arginine-vasopressin (AVP) potentiated the ischemic decrease of the CA1 presynaptic potential. In contrast, a V, receptor antagonist produced a dose-dependent neuroprotective effect, whereas a V, receptor antagonist had no effect. The AVP-induced decrease of the CAI presynaptic potential was completely blocked by simultaneous application of the V, receptor antagonist. Because AVP(4-9) is regarded as the major proteolytic product of AVP in the rat brain, we examined its effect on the ischemic decrease of the CA1 presynaptic potential. Treatment with AVP(4-9) produced a more marked reduction of the potential than treatment with AVP itself. The present study demonstrates that stimulation of the V-1, receptor has a detrimental effect on the development of ischemic damage whereas V-1, receptor blockade has a neuroprotective effect, suggesting that AVP may potentiate ischemic neuronal deficits via V-1 receptor stimulation.

    DOI

  • AGING IMPAIRS METHAMPHETAMINE-INDUCED FREE-RUNNING AND ANTICIPATORY LOCOMOTOR-ACTIVITY RHYTHMS IN RATS

    S SHIBATA, Y MINAMOTO, M ONO, S WATANABE

    NEUROSCIENCE LETTERS   172 ( 1-2 ) 107 - 110  1994.05

     View Summary

    In the present experiment, under drinking of 0.01% methamphetamine (MA), young rats exhibited free-running locomotor rhythm ranging from 24.6 to 30.9 h period under light-dark cycle. In contrast, 24-month-old rats showed a low activity and did not exhibit MA-induced free-running rhythm. When MA was injected at a fixed time of day (14:00), young rats exhibited intense locomotor activity from 1-2 h before drug-injection time. This preinjection activity was still detected on the withdrawal day. In addition, intense locomotor activity was observed for 12:00-17:00, even on the withdrawal day (MA-associated anticipatory activity). Thus, MA-associated activity of the withdrawal day became an another good index for MA anticipatory activity rhythms whereas old rats did not exhibit this anticipatory activity. The present study suggests that aging strongly impairs the manifestation of MA-induced free-running and anticipatory activity rhythms in rats.

    DOI

  • EFFECTS OF NITRIC-OXIDE SYNTHASE INHIBITORS ON N-METHYL-D-ASPARTATE-INDUCED PHASE DELAY OF CIRCADIAN-RHYTHM OF NEURONAL-ACTIVITY IN THE RAT SUPRACHIASMATIC NUCLEUS IN-VITRO

    A WATANABE, T HAMADA, S SHIBATA, S WATANABE

    BRAIN RESEARCH   646 ( 1 ) 161 - 164  1994.05

     View Summary

    Excitatory amino acid (EAA) receptors such as N-methyl-D-aspartate (NMDA) and non-NMDA receptors have been suggested to play an important role in the regulation of photic information from the retina to the suprachiasmatic nucleus (SCN). Therefore, we investigated the role of glutamate as a retinohypothalamic transmitter by analyzing the phase-resetting effects of NMDA and a non-NMDA agonist, (R,S)-alpha-amino-3-hydroxy-5-methylisoxazo-4-propionic acid (AMPA), on the circadian rhythm of SCN firing activity. Nitric oxide (NO) production is believed to be an essential intermediate in NMDA-induced cGMP production in the CNS. Thus, we examined the effects of blockers of NO production on NMDA- or AMPA-induced phase delay of SCN activity rhythm. N-nitro-L-arginine methylester (L-NAME) blocked NMDA- but not AMPA-induced phase shift, indicating the involvement of NO synthesis in NMDA-induced phase changes. L-arginine but not D-arginine caused a phase delay, and L-NAME blocked L-arginine-induced phase delay. In addition, cotreatment with NMDA and L-arginine did not have an additive effect. These results suggest that NO production itself is involved in the phase change of SCN neuron activity, and NMDA-induced phase changes are also mediated via activation of NO synthesis in this nucleus.

    DOI

  • NEUROPROTECTIVE EFFECT OF CHOLECYSTOKININ(B) RECEPTOR ANTAGONIST ON ISCHEMIA-INDUCED DECREASE IN CA1 PRESYNAPTIC FIBER SPIKES IN RAT HIPPOCAMPAL SLICES

    Y MINAMOTO, T TANAKA, S SHIBATA, S WATANABE

    NEUROSCIENCE LETTERS   167 ( 1-2 ) 81 - 84  1994.02

     View Summary

    The effects of cholecystokinin (CCK) receptor agonists and antagonists on hypoxia/hypoglycemia (ischemia)-induced decrease in CA1 presynaptic fiber spikes elicited by the stimulation of Schaffer collaterals were investigated using rat hippocampal slices. Treatment with the CCKB receptor agonist CCK tetrapeptide (CCK4, 0.01-10 mu M) exacerbated the ischemia-induced decrease in the CA1 presynaptic potential in a concentration-dependent manner. Whereas, treatment with the CCKB receptor antagonist [(3R(+)-N-(2,3-dihydro-1-methyl-2-oxo-5-phenyl-1H-1,4-benzodiazepin-3-yl)-N1-(3-methylphenyl)-urea] (L365260), and not with CCKA receptor antagonist [(3S(-)-N-(2,3-dihydro-1-methyl-2-oxo-5phenyl-1H-1,4-benzodiazepin-3-yl)-1H-indole-2-carboxamide] (L364718), produced a concentration-dependent attenuation of the ischemia-induced decrease. The magnitude of recovery of the CAI field potentials in L365260-treated groups at 10 and 100 nM was 34% and 45%, respectively. The neuroprotective effect of L365260 (0.01 and 0.1 mu M) was completely blocked by co-treatment with CCK4 (0.1 mu M), a concentration that did not affect the decreased presynaptic potential induced by ischemia. These results demonstrated that the stimulation of the CCKB receptor played a detrimental role in the development of ischemic damage, whereas the blockade of CCKB receptors played a neuroprotective role in ischemic damage, suggesting a facilitatory role of CCK receptor-operated function in ischemia-induced neuronal deficits.

    DOI

  • PROTEIN-SYNTHESIS INHIBITOR BLOCKS (R,S)-ALPHA-AMINO-3-HYDROXY-5-METHYLISOXAZOLE-4-PROPIONIC ACID (AMPA)-INDUCED OR SUBSTANCE-P-INDUCED PHASE-SHIFT OF THE CIRCADIAN-RHYTHM OF NEURONAL-ACTIVITY IN THE RAT SUPRACHIASMATIC NUCLEUS IN-VITRO

    S SHIBATA, A WATANABE, T HAMADA, S WATANABE

    NEUROSCIENCE LETTERS   168 ( 1-2 ) 159 - 162  1994.02

     View Summary

    The mammalian suprachiasmatic nucleus (SCN) has been identified as a circadian pacemaker. N-methyl-D-aspartate (NMDA), non-NMDA and substance P receptors have been suggested to be involved in handling: of photic information in the SCN. In the Aplysia eyes, in which the circadian clocks are involved, serotonin- or cAMP-induced phase changes of the circadian rhythm were reported to be blocked by protein-synthesis inhibitors. Therefore, we investigated whether protein-synthesis inhibitor can block the non- NMDA receptor agonist (R,S)-alpha-amino-3-hydroxy-5-methylisoxazole-4-propionic acid hydrobromide (AMPA)- or substance P (SP)-induced phase changes of SCN activity rhythm. Although application of 10 mu M cycloheximide alone during the early part of the subjective night did not cause phase change, it blocked both 10 mu M AMPA- and 1 mu M SP-induced phase delay. The present result suggests that protein synthesis may be required in the manifestation of AMPA- and SP-induced phase change of circadian clock.

    DOI

  • 哺乳動物におけるサーカディアンリズムの薬理

    柴田重信, 渡辺繁紀

    薬学雑誌   114 ( 9 ) 637 - 654  1994

    DOI

  • サーカディアンリズムの体内時計としての視床下部視交叉上核

    柴田重信, 渡辺繁紀治療

    治療学   28   21 - 24  1994

  • A FACILITATORY ROLE OF VASOPRESSIN IN HYPOXIA/HYPOGLYCEMIA-INDUCED IMPAIRMENT OF DOPAMINE RELEASE FROM RAT STRIATAL SLICES

    M OCHI, S KOIZUMI, S SHIBATA, S WATANABE

    BRAIN RESEARCH   633 ( 1-2 ) 91 - 96  1994.01

     View Summary

    The excitatory amino acid, glutamate plays a crucial role in the pathogenesis of brain damage caused by anoxia and/or hypoglycemia. Although vasopressin (VP) also acts as an excitatory transmitter in the CNS, little is known about its effect on hypoxic and/or ischemic brain damage. In this study, we investigated the effect of arginine vasopressin (AVP) on hypoxia/ hypoglycemia-induced impairment of dopamine release from striatal slices. Striatal slices were incubated in hypoxia-/ hypoglycemia-inducing medium with or without AVP (0.01-1.0 mu M) for 20 min. After 1-3 h of washout in normal medium, high K+-evoked dopamine release from the slices were examined. Hypoxia/hypoglycemia-induced decrease of striatal dopamine release was reversed by the removal of Ca2+ in the medium, but not by VP1- or VP2-receptor antagonist. In contrast, AVP potentiated the hypoxia/hypoglycemia-induced decrease of dopamine release in the striatum. This AVP-induced deterioration of the striatal response was antagonized by VP2 receptor antagonist, but not by VP1 receptor antagonist. The present results suggest that AVP may play a facilitatory role in hypoxia/hypoglycemia-induced dopamine release deficit mediated through the activation of VP2 receptor.

    DOI

  • Calmodulin Inhibitors Produce Phase Shifts of Circadian Rhythms In Vivo and In Vitro

    Shigenobu Shibata, Robert Y. Moore

    Journal of Biological Rhythms   9 ( 1 ) 27 - 41  1994

     View Summary

    The effect of calmodulin inibitors on the circadian rhythm of locomotor activity and on the rhythm of suprachiasmatic nuclear (SCN) neuron firing rate recorded in vitro from hypothalamic slices was examined. Trifluoperazine produces changes in a dose-dependent manner in the phase of the activity rhythm, with phase advances throughout most of the subjective day extending into the subjec tive night. These phase changes in the activity rhythm occur rapidly and without induction of locomotor activity at the time of treatment. Similarly, trifluoperazine and the naphthalenesulfonamide W-7 produce changes in phase delays in the subjective night extending into early subjective day. The effects are greater with respect to amplitude when measured acutely after treatment than in the next cycle, and both the acute and next-day effects are greater than those observed in vivo, indicating that data from in vitro studies need to be interpreted with caution. These observations indicate that calmodulin inhibitors affect rhythms directly in vivo by altering SCN neuron pacemaker function, as this reflects involvement of calcium-calmodulin binding with activation of a calmodulin-dependent kinase, either to alter intracellular cAMP levels or to alter gene expression directly to modulate the phase of the SCN clock. © 1994, Sage Publications. All rights reserved.

    DOI PubMed

  • N-methyl-D-aspartate induces phase shifts of the circadian rhythm of neuronal activity of the rat suprachiasmatic nucleus in vitro.

    Shibata S, Watanabe A, Hamada T, Ono M, Watanabe S

    Am J Physiol   267 ( 2 Pt 2 ) R360 - R364  1994

  • Involvement of vasoactive intestinal polypeptide in NMDA-induced phase delay of firing activity rhythm in the suprachiasmatic nucleus in vitro

    Shigenobu Shibata, Michiko Ono, Keiko Tominaga, Toshiyuki Hamada, Akihito Watanabe, Shigenori Watanabe

    Neuroscience and Biobehavioral Reviews   18 ( 4 ) 591 - 595  1994

     View Summary

    Glutamate has been reported to be involved in the transmission of photic information from the retina to the suprachiasmatic nucleus (SCN). Therefore, we investigated whether the application of N-methyl-d-aspartate (NMDA), a glutamate receptor agonist could, reset the circadian rhythm of SCN firing activity in vitro. Treatment with NMDA for 1 h between projected zeitgeber time (ZT) 13-14 produced a phase delay in a concentration-dependent manner. The NMDA-induced phase delay was antagonized by an NMDA-receptor antagonist, MK-801 (100 μM). The retinohypothalamic tract has been reported to make terminals on neurons possessing vasoactive intestinal polypeptide (VIP). Therefore, we investigated the effects of NMDA on VIP release from the SCN and on VIP immunoreactivity in the SCN. Application of NMDA for 15 min between ZT 13-15 increased release of VIP from the SCN. In contrast to release, the content of VIP in the SCN tissue was reduced by application of NMDA. Immunohistochemical analysis revealed that application of NMDA for 4 h or 1 h reduced VIP immunoreactivity in the SCN. To investigate the possibility that VIP released by NMDA could reset SCN neuronal activity, we examined the effects of VIP on the SCN neuronal activity rhythm. Cotreatment with VIP (1 μM) and gastrin-releasing peptide (1 μM) for 1 h between ZT 13-14 caused a phase-delay of SCN activity rhythm. These findings suggest that activation of NMDA receptors during early subjective night causes a phase delay of the SCN neuronal activity via facilitation of VIP release in this nucleus.

    DOI PubMed

  • GABA(A) RECEPTOR AGONIST MUSCIMOL CAN RESET THE PHASE OF NEURAL ACTIVITY RHYTHM IN THE RAT SUPRACHIASMATIC NUCLEUS IN-VITRO

    K TOMINAGA, S SHIBATA, T HAMADA, S WATANABE

    NEUROSCIENCE LETTERS   166 ( 1 ) 81 - 84  1994.01

     View Summary

    We investigated the phase-resetting effect of muscimol, gamma-amino butyric acid (GABA)(A) receptor agonist, on the circadian neural activity rhythm of the rat suprachiasmatic nucleus (SCN), which contains a circadian pacemaker. Acute application of muscimol inhibited the neural activity of the SCN in a dose-dependent manner. Under the tissue culture condition, the treatment with 10 mu M muscimol during the early- to mid-subjective day on the first day (day 1) in vitro produced the largest phase advance in neural activity rhythm of the SCN on day 2. By contrast, the administration of muscimol during the subjective night produced no change. These phase changes were similar to those reported for dark pulses in constant light. These findings indicate that muscimol can directly affect SCN neurons and reset the circadian pacemaker in the SCN. The GABA neural function through the activation of GABA(A) receptors may play a role in modulating the phase of the SCN clock, especially during the subjective day.

    DOI

  • N-methyl-D-aspartate induces phase shifts in circadian rhythm of neuronal activity of rat SCN in vitro

    S. Shibata, A. Watanabe, T. Hamada, M. Ono, S. Watanabe

    American Journal of Physiology - Regulatory Integrative and Comparative Physiology   267 ( 2 ) R360 - R364  1994  [Refereed]

     View Summary

    The suprachiasmatic nucleus (SCN) acts as a pacemaker for mammalian circadian rhythms. Receptors for excitatory amino acids like N-methyl-D- aspartate (NMDA) and non-NMDA receptors have both been found to play an important role in the transmission of photic information from the retina to the SCN. Therefore, we investigated whether the application of glutamate receptor agonists could reset the phase of the circadian rhythm of SCN firing activity in vitro. Treatment with NMDA (0.1-10 μM) for 15 min or 1 h during the early part of the subjective night produced phase delay, whereas treatment during the late subjective night caused an advance in phase. The phase-response curve for NMDA was similar to that previously obtained in response to light pulses in vivo. Application of DL-α-amino-3-hydroxy-5- methylisoxazole-4-propionic acid hydrobromide (AMPA) (1 or 10 μM), a non- NMDA-receptor agonist, also produced a dose-dependent phase delay of SCN activity. The NMDA-induced phase delay was antagonized by an NMDA-receptor antagonist MK-801. These findings suggest that both NMDA and non-NMDA receptors may be involved in the transmission of information to the SCN via the retinohypothalamic tract. In addition, both the advances and delays in phase caused by NMDA were potentiated by cotreatment with neuropeptide Y, whereas AMPA-induced phase delay was not potentiated by neuropeptide Y. This points to a functional link between NMDA and neuropeptide Y receptor-mediated mechanisms in the SCN.

    PubMed

  • EFFECT OF SOMATOSTATIN ON CIRCADIAN-RHYTHMS OF FIRING AND 2-DEOXYGLUCOSE UPTAKE IN RAT SUPRACHIASMATIC SLICES

    T HAMADA, S SHIBATA, A TSUNEYOSHI, K TOMINAGA, S WATANABE

    AMERICAN JOURNAL OF PHYSIOLOGY   265 ( 5 ) R1199 - R1204  1993.11

     View Summary

    In mammals, the suprachiasmatic nucleus (SCN) of the hypothalamus appears to act as a circadian clock. The SCN vasoactive intestinal polypeptide-like immunoreactive neurons, which may act to mediate photic information in the SCN, receive input from neurons immunoreactive for somatostatin (SST). Therefore we investigated the role of SST as a transmitter for entrainment by analyzing the phase-resetting effect of SST on the circadian rhythm of SCN firing activity. Perfusion of SST increased 2-deoxyglucose uptake at circadian time (CT) 18, but not at CT6. A 1-h or 15-min treatment with SST produced phase delays when it was administered at CT13-14 and phase advances at CT22-23. Thus SST-induced phase changes are similar to those for light pulses to animals under constant darkness. The present findings suggest that SST is a transmitter for mediating information of entrainment to circadian clocks within the SCN.

  • Neuropeptide Y and optic chiasm stimulation of affect suprachiasmatic nucleus circadian function in vitro

    Shigenobu Shibata, Robert Y. Moore

    Brain Research   615 ( 1 ) 95 - 100  1993.06

     View Summary

    The retinohypothalamic tract (RHT) is a direct pathway from the retina to the suprachiasmatic nucleus (SCN). Electrical stimulation of the optic nerve or optic chiasm activates the RHT and produces shifts in phase of a circadian rhythm in SCN neuron activity in rat hypothalamic slices in vitro. The phase response curve (PRC) for this effect is very similar to that obtained from administration of light pulses to intact animals maintained in constant darkness. The effect of optic chiasm stimulation is blocked by tetrodotoxin. In addition to the RHT, there is a second entraining pathway, the geniculohypothalamic tract, which arises from neuropeptide Y (NPY)-containing neurons of the intergeniculate leaflet of the lateral geniculate complex. In contrast to optic chiasm stimulation, NPY produces phase shifts in the rhythms of SCN neuron firing rate in vitro with a PRC that similar to that for NPY infusion into the SCN in intact animals as well as that produced by a series of treatments that induce locomotor activity. These results indicate that phase shifts of the circadian rhythm of SCN neuron activity may be produced by activation of two different entraining pathways and that the physiological actions of these pathways on pacemaker function are markedly different. © 1993.

    DOI PubMed

  • FACILITATORY EFFECT OF PHORBOL ESTER ON 2-DEOXYGLUCOSE UPTAKE IN RAT HIPPOCAMPAL SLICES

    S SHIBATA, A NAKASHIO, S UEKI, S WATANABE

    EUROPEAN JOURNAL OF PHARMACOLOGY-MOLECULAR PHARMACOLOGY SECTION   245 ( 3 ) 257 - 261  1993.05

     View Summary

    It is well known that synaptic potentiation in the hippocampus can be produced by phorbol ester, a protein kinase C activator. The 2-deoxyglucose uptake is an index of regional glucose utilization which predominantly reflects activity in the axonal terminal of neuronal pathways. In the present experiment, therefore, we examined whether application of phorbol ester produces a facilitatory effect on 2-deoxyglucose uptake by the rat hippocampus in vitro. The application of phorbol-12,13-dibutyrate (PdBU) produced an elevation of 2-deoxyglucose uptake, while pretreatment with PdBU for 60 min eliminated the pdBU-induced elevation. Pretreatment with protein kinase C inhibitors, K252a (0.1 and 1 muM) or staurosporine (0.1 and 1 muM), was found to block significantly the PdBU-induced elevation of 2-deoxyglucose uptake. In addition, the facilitatory effect of glutamate, quisqualate and carbachol on 2-deoxyglucose uptake was reduced by pretreatment with PdBU. In the present experiment, we demonstrated that application of phorbol ester caused an elevation of 2-deoxyglucose uptake, which is linked in turn to neuronal activity, suggesting a positive relationship between protein kinase C activation and energy consumption.

    DOI

  • WORKING MEMORY DEFICITS FOLLOWING MUSCARINIC BLOCKADE COMBINED WITH DEPLETION OF BRAIN SOMATOSTATIN IN RATS

    M OHNO, S SHIBATA, T YAMAMOTO, S WATANABE

    BRAIN RESEARCH   610 ( 2 ) 348 - 353  1993.05

     View Summary

    In a working memory task with three-panel runway paradigm, cysteamine, a depletor of somatostatin, at 100 or 200 mg/kg i.p. given 24 h before testing, had no effect on the number of errors (attempts to pass through two incorrect panels of the three panel-gates at four choice points). Cysteamine at 100 mg/kg caused a significant reduction in somatostatin-like immunoreactivity in the rat brain, including the hippocampus and cerebral cortex. Working memory errors were significantly increased by scopolamine, a muscarinic receptor antagonist, at 0.32 mg/kg i.p. given 20 min before testing, whereas errors were not affected by the 0.1 mg/kg dose. Combined administration of 100 mg/kg cysteamine and 0.1 mg/kg scopolamine significantly increased the number of working memory errors. However, cysteamine at 100 mg/kg and scopolamine at 0.1 mg/kg had no effect on reference memory errors, whether they were administered alone or in combination. These results suggest that depletion of brain somatostatin aggravates working memory deficits induced by blockade of muscarinic receptors.

    DOI

  • TETRODOTOXIN DOES NOT AFFECT CIRCADIAN-RHYTHMS IN NEURONAL-ACTIVITY AND METABOLISM IN RODENT SUPRACHIASMATIC NUCLEUS INVITRO

    S SHIBATA, RY MOORE

    BRAIN RESEARCH   606 ( 2 ) 259 - 266  1993.03

     View Summary

    Single unit activity of suprachiasmatic nucleus (SCN) neurons recorded from hamster and rat hypothalamic slices in vitro exhibits a circadian rhythm in firing rate. Tetrodotoxin (TTX) produces electrical silence during the period of administration but has no effect on the oscillatory activity of the circadian pacemaker in both hamster and rat SCN. TTX does not affect either the phase of the rhythm in firing rate at any time of the circadian day, or that of the rhythm in glucose metabolism as demonstrated by the 2-deoxyglucose method. These data are in accord with the view that SCN neuron firing rate and glucose utilization is an expression of pacemaker activity but that pacemaker function is maintained in the SCN independent of neuronal function manifested by sodium dependent action potentials.

    DOI

  • A NEUROPROTECTIVE EFFECT OF HISTAMINE H-1-RECEPTOR ANTAGONIST ON ISCHEMIA-INDUCED DECREASE IN 2-DEOXYGLUCOSE UPTAKE IN RAT HIPPOCAMPAL SLICES

    S SHIBATA, S WATANABE

    NEUROSCIENCE LETTERS   151 ( 2 ) 138 - 141  1993.03

     View Summary

    The effects of histamine (HA) receptor antagonists on hypoxia + hypoglycemia (ischemia)-induced impairment of 2-deoxyglucose (2-DG) uptake by rat hippocampal slices was evaluated. Hippocampal slices were exposed to 20-min ischemia and then returned to oxygenated and glucose-containing Krebs-Ringer solution for 6 h. Ischemia reduced 2-DG uptake in the hippocampal slices. The ischemia-induced reduction in 2-DG uptake was attenuated by pretreatment with H-1 receptor antagonist but not with H-2 receptor antagonist. Treatment with HA exacerbated the ischemia-induced decrease. The H-1 receptor antagonist-induced neuroprotective effect was blocked by co-treatment with HA. The present study suggests that the blockade of H-1 receptor-mediated function has a protective role in ischemia-induced decreases in glucose metabolism in hippocampal slices.

    DOI

  • FACILITATION OF 2-DEOXYGLUCOSE UPTAKE IN RAT CORTEX AND HIPPOCAMPUS SLICES BY SOMATOSTATIN IS INDEPENDENT OF CHOLINERGIC ACTIVITY

    S SHIBATA, Y KOGA, T HAMADA, S WATANABE

    EUROPEAN JOURNAL OF PHARMACOLOGY   231 ( 3 ) 381 - 388  1993.02

     View Summary

    2-Deoxyglucose (2-DG) uptake is an index of regional glucose utilization which reflects predominantly activity in the axonal terminal of neuronal pathways. The present experiments showed that somatostatin elevated 2-DG uptake in rat cortex and hippocampus slices. Treatment with somatostatin-14 and somatostatin-28 markedly enhanced 2-DG uptake, whereas the amino-terminal fragment of somatostatin-28 did so only slightly. This effect appeared to be mediated by an interaction with somatostatin receptors because cyclo-somatostatin, a somatostatin antagonist, abolished the effect of somatostatin-14. The increase in 2-DG uptake caused by somatostatin-14 was blocked by the calcium channel antagonist, nifedipine, but not by tetrodotoxin, suggesting that the action of somatostatin does not require the initiation of impulse activity. somatostatin enhanced the KCl-induced release of acetylcholine, suggesting that a cholinergic mechanism is involved in the somatostatin-induced cellular responses. We therefore examined whether acetylcholine receptor antagonists block the somatostatin-induced increase in 2-DG uptake. Neither muscarinic nor nicotinic receptor antagonists affected the somatostatin-14-induced response. The present results suggest that somatostatin has a stimulatory effect on energy metabolism and that this effect is independent of acetylcholine receptor mechanism.

    DOI

  • IMPROVING EFFECT OF ACETYLCHOLINE-RECEPTOR AGONISTS ON A DEFICIT OF 2-DEOXYGLUCOSE UPTAKE IN CEREBRAL CORTICAL AND HIPPOCAMPAL SLICES IN AGED AND AF64A-TREATED RATS

    S SHIBATA, K KODAMA, Y KOGA, S UEKI, S WATANABE

    BRAIN RESEARCH   603 ( 2 ) 248 - 254  1993.02

     View Summary

    The aim of the present study was to determine whether the facilitation of 2-deoxyglucose (2-DG) uptake in the cerebral and hippocampal slices by nicotinic and muscarinic receptor agonists is compromised in the aged rat brain. For this, the effects of the nicotinic receptor agonist nicotine, the muscarinic receptor agonists oxotremorine and McN-A-343, and the ACh esterase inhibitors physostigmine and NK247 on 2-DG uptake in the brain slices of young (2-month-old) and aged (24-26-month-old) rats were tested. The decrements of 2-DG uptake in the cortical slices of aged rats were significantly attenuated by treatment with oxotremorine, nicotine and amiridine. In contrast, the metabolic responsivity of hippocampal slices to these drugs was reduced. To assess whether age-related changes in 2-DG uptake may be due to deficits in cholinergic function, we tested these drugs on the decrements of 2-DG uptake in ethylcholine aziridinium (a neurotoxic analog of choline) injected rats. The reductions of 2-DG uptake by injection of ethylcholine aziridinium was attenuated by oxotremorine but not by physostigmine. The present results reveal that metabolic decrements in the cerebral cortex from aged or ethylcholine aziridinium-injected rats were attenuated by muscarinic and nicotinic receptor agonists, suggesting that the muscarinic and nicotinic receptor mechanism in the cerebral cortex may be involved in cholinergic drug-induced functional recovery in aged rats.

    DOI

  • Anxiolytic activities of pentanthrene type heterocyclic compounds

    T. Imamura, A. Noda, S. Shibata, S. Watanabe, H. Noda, Y. Ono, S. Goto, Y. Inoue

    Yakugaku Zasshi   113 ( 5 ) 400 - 405  1993

     View Summary

    A series of pentanthrene type heterocyclic compounds were synthesized and evaluated for anxiolytic activities by three kinds of behavioral pharmacological tests. Several compounds showed anxiolytic activities. In particular, s-triazolo[3,4-a]phthalazine (Tri-P) and 3-propyl derivatives of Tri-P(PTP) showed remarkable activities, although the activities were slightly lower than those of diazepam. The results suggested that Tri-P or PTP is a useful lead compound for the development of the antianxietic agents. The relationship between the structure and anxiolytic activity, and the inducing mechanism of the activity was discussed.

    DOI PubMed

  • ISCHEMIA-INDUCED IMPAIRMENT OF 2-DEOXYGLUCOSE UPTAKE AND CA1 FIELD POTENTIALS IN RAT HIPPOCAMPAL SLICES - PROTECTION BY 5-HT(1A) RECEPTOR AGONISTS AND 5-HT(2) RECEPTOR ANTAGONISTS

    S SHIBATA, Y KAGAMIISHI, K TOMINAGA, K KODAMA, S UEKI, S WATANABE

    EUROPEAN JOURNAL OF PHARMACOLOGY   229 ( 1 ) 21 - 29  1992.12

     View Summary

    Various in vitro models have been developed to study ischemia and/or hypoxia. In the present experiment, we examined whether hypoxia/hypoglycemia (ischemia) in rat hippocampal slices reduced the 2-deoxyglucose (2-DG) uptake and CA1 field potentials evoked by stimulation of Schaffer collaterals. Autoradiograms revealed that ischemia for 15 or 20 min reduced 2-DG uptake in the stratum radiatum of the CA1 and the dentate gyrus. Similarly, the CA1 field potentials of slices exposed to ischemia for 15 and 20 min decreased by about 70 and 90% after a 6-h washout. In the second experiment, we evaluated the neuroprotective effect of the 5-HT1A receptor agonists 8-OH-DPAT and buspirone, and the 5-HT2 receptor antagonists cyproheptadine, mianserin and ketanserin on deficits of 2-DG uptake and Schaffer-CA1 field potentials induced by ischemia. The 5-HT1A receptor agonists and 5-HT2 receptor antagonists exhibited significant neuroprotective actions against ischemia-induced deficits. Therefore, impairments of 2-DG uptake and CA1 field potentials induced by ischemia may be good markers of ischemia-induced functional deficits. The attenuating action of 5-HT1A receptor agonists and 5-HT2 receptor antagonists were assessed using this model of ischemia.

    DOI

  • EFFECT OF SUBSTANCE-P ON CIRCADIAN-RHYTHMS OF FIRING ACTIVITY AND THE 2-DEOXYGLUCOSE UPTAKE IN THE RAT SUPRACHIASMATIC NUCLEUS INVITRO

    S SHIBATA, A TSUNEYOSHI, T HAMADA, K TOMINAGA, S WATANABE

    BRAIN RESEARCH   597 ( 2 ) 257 - 263  1992.12

     View Summary

    The suprachiasmatic nuclei (SCN) have been identified as a pacemaker for many circadian rhythms in mammals. Although substance P (SP) fibers from retina are found to terminate the SCN, the physiological role of this peptide is uncertain. The 2-deoxyglucose (2-DG) uptake and firing activity in the SCN show a robust circadian change. SP causes an increase in 2-DG uptake by SCN during the subjective night but not during subjective day. SP-induced increase in 2-DG uptake is blocked by co-treatment with the SP receptor antagonist, spantide. Treatment with SP produces phase shifts of circadian rhythm in spontaneous neural activity in SCN neurons with a phase-response curve that is similar to the effect of light pulses to animals under constant darkness. SP-induced phase change is also blocked by pretreatment with spantide. SP-induced increase in 2-DG uptake and phase changes in firing activity occur only during subjective night, at circadian times when photic phase shifting of activity occurs. The present results suggest that SP may be an important transmitter for conveying environmental light-dark information from retina to the SCN.

    DOI

  • NEUROPROTECTIVE EFFECT OF 5-HT(3) RECEPTOR ANTAGONIST ON ISCHEMIA-INDUCED DECREASE IN CA1 FIELD POTENTIAL IN RAT HIPPOCAMPAL SLICES

    Y KAGAMIISHI, S SHIBATA, S WATANABE

    EUROPEAN JOURNAL OF PHARMACOLOGY   224 ( 1 ) 51 - 56  1992.11

     View Summary

    The effect of 5-HT3 receptor agonists and antagonists on the hypoxia/hypoglycemia (ischemia)-induced decrease in CA1 field potential elicited by stimulation of Schaffer collaterals was investigated using rat hippocampal slices. Treatment with the 5-HT3 receptor agonist, 2-methyl-5-HT (1-10 muM), exacerbated the ischemia-induced decreased in CA1 field potential, whereas treatment with the 5-HT3 receptor antagonist, Y-25130 (0.1-100 muM), or the 5-HT2 receptor antagonist, ketanserin (10, 100 muM), produced dose-dependent neuroprotection against the ischemia-induced decrease. However, in normal non-ischemic solution these treatments did not significantly change the CA1 field potential. The protective action of Y-25130 was blocked by co-treatment with 2-methyl-5-HT. The magnitude of protection in the Y-25130-treated group (EC50, 1.8 muM) was about 20 times greater than that in the ketanserin-treated group (EC50, 33 muM). The present study demonstrated that stimulation of 5-HT3 receptors plays a detrimental role in the development of ischemic damage, whereas blockade of the 5-HT3 receptor plays a neuroprotective role in ischemic damage, suggesting a facilitatory role of 5-HT neurons in ischemia-induced neuronal deficits.

    DOI

  • EFFECT OF MUSCARINIC CHOLINERGIC DRUGS ON ISCHEMIA-INDUCED DECREASES IN GLUCOSE-UPTAKE AND CA1 FIELD POTENTIALS IN RAT HIPPOCAMPUS SLICES

    S SHIBATA, K KODAMA, K TOMINAGA, T TANAKA, S WATANABE

    EUROPEAN JOURNAL OF PHARMACOLOGY   221 ( 1 ) 113 - 119  1992.10

     View Summary

    To clarify the role of muscarinic acetylcholine receptors in the hypoxia/ hypoglycemia (ischemia)-induced functional deficit in hippocampal neurons, we examined the effect of cholinergic drugs on ischemia-induced impairments of glucose uptake and CA1 field potentials in hippocampus slices. Muscarinic receptors were subdivided into M1 (high affinity for pirenzepine) and M2 (low affinity for pirenzepine) subtypes. The M1 receptor subtype is coupled to an increase in phosphoinositide hydrolysis and the M2 receptor subtype is associated with inhibition of adenylate cyclase. The greater potency of carbachol in stimulating phosphoinositide hydrolysis resulted in exacerbated ischemia-induced deficits. Treatment with the muscarinic receptor antagonists scopolamine and pirenzepine (M1 receptor-selective antagonist) had a strong dose-dependent protective effect against ischemia-induced deficits. Oxotremorine and McN-A-343, weak stimulators of phosphoinositide hydrolysis and strong inhibitors of adenylate cyclase, had a weak neuroprotective action against ischemia-induced deficits. These results suggest that stimulation of M1 musarinic receptors coupled with an increase in phosphoinositide hydrolysis may play a facilatory role in ischemia-induced deficits. Stimulation of M2 muscarinic receptors may play an inhibitory role in ischemia-induced neuronal deficits.

    DOI

  • ASSESSMENT OF THE ROLE OF ADRENOCEPTOR FUNCTION IN ISCHEMIA-INDUCED IMPAIRMENT OF 2-DEOXYGLUCOSE UPTAKE AND CA1 FIELD POTENTIAL IN RAT HIPPOCAMPAL SLICES

    S SHIBATA, K KODAMA, K TOMINAGA, S UEKI, S WATANABE

    EUROPEAN JOURNAL OF PHARMACOLOGY   221 ( 2-3 ) 255 - 260  1992.10

     View Summary

    The release of catecholamines, dopamine and noradrenaline has been suggested to play a role in mediating ischemic damage in susceptible brain regions, the hippocampus and striatum. We now provide evidence that suggests a role for adrenoceptors in the deficit of 2-deoxyglucose uptake and CA1 field potential induced in hippocampal slices by hypoxia/hypoglycemia (ischemia). Treatment with alpha1- or beta-adrenoceptor agonists or cAMP potentiated an ischemia-induced decline of both 2-deoxyglucose uptake and CA1 field potential in hippocampal slices, whereas alpha1- or beta-adrenoceptor antagonists, or alpha2-adrenoceptor agonists produced a remarkable neuroprotective action against these deficits. The results indicate that stimulation of adrenoceptors may play a detrimental role in the development of ischemic damage, and suggest a neuroprotective action for adrenoceptor antagonists, which may lessen the functional deficits induced by ischemia.

    DOI

  • NEUROPROTECTIVE EFFECT OF PROTEIN-KINASE-C INHIBITORS ON OXYGEN GLUCOSE FREE-INDUCED DECREASES IN 2-DEOXYGLUCOSE UPTAKE AND CA1 FIELD POTENTIALS IN RAT HIPPOCAMPAL SLICES

    S SHIBATA, K TOMINAGA, Y KAGAMIISHI, S WATANABE

    BRAIN RESEARCH   594 ( 2 ) 290 - 294  1992.10

     View Summary

    Staurosporine, a protein kinase C inhibitor, was found to produce a neuroprotective effect against an ischemic insult in both gerbils and rats in vivo. We have demonstrated that rat hippocampal slices exposed to oxygen/glucose-free medium showed decreases in 2-deoxyglucose (2-DG) uptake and CA1 field potentials elicited by the stimulation of Schaffer collaterals. Therefore we examined the effect of protein kinase C inhibitors on oxygen/glucose free-induced impairments of 2-DG uptake and CA1 field potentials. Pretreatment with staurosporine, K252a and H-7 attenuated decreases in 2-DG uptake and CA1 field potentials. Treatment with phorbol ester, a protein kinase C activator, for a long period (90 min) was found to induce a down-regulation of protein kinase C activity. Therefore we examined the effect of pretreatment with phorbol ester for 90 min on oxygen/glucose free-induced decreases in 2-DG uptake and CA1 field potentials. These decrements were not attenuated by 5-min treatment with phorbol ester but were attenuated by 90-min treatment. The present results suggest that the treatment which decreases protein kinase C activity shows a neuroprotective action against oxygen/glucose free-induced deficits of metabolic and synaptic activity in hippocampal slices.

    DOI

  • A NEUROPROTECTIVE EFFECT OF ADENOSINE-A1-RECEPTOR AGONISTS ON ISCHEMIA-INDUCED DECREASE IN 2-DEOXYGLUCOSE UPTAKE IN RAT HIPPOCAMPAL SLICES

    K TOMINAGA, S SHIBATA, S WATANABE

    NEUROSCIENCE LETTERS   145 ( 1 ) 67 - 70  1992.09

     View Summary

    The effects of adenosine (A) receptor agonists on ischemia-induced impairment of 2-deoxyglucose (2-DG) uptake by rat hippocampal slices was evaluated. Hippocampal slices were exposed to 20-min hypoxia + hypoglycemia (ischemia) and then returned to oxygenated and glucose-containing Krebs-Ringer solution for 6 h. Ischemia reduced 2-DG uptake in the hippocampal slices. The ischemia-induced reduction in 2-DG uptake was attenuated by pretreatment with A1 receptor agonists but not with A2 receptor agonists. 8-Phenyltheophylline, an A1 receptor antagonist, "acerbated the ischemia-induced decrease. The A1 receptor agonist-induced neuroprotective effect was blocked by co-treatment with 8-phenyltheophylline. The present study suggests that the A1 receptor-mediated function has a protective role in ischemia-induced decreases in glucose metabolism in hippocampal slices.

    DOI

  • PROTECTIVE EFFECT OF MINAPRINE AGAINST THE ABNORMAL CHANGES OF 2-DEOXYGLUCOSE UPTAKE BY RAT HIPPOCAMPAL SLICES INDUCED BY HYPOXIA HYPOGLYCEMIA

    K KODAMA, S SHIBATA, S UEKI

    JAPANESE JOURNAL OF PHARMACOLOGY   60 ( 1 ) 33 - 38  1992.09

     View Summary

    Effect of minaprine on hypoxia- or hypoxia/hypoglycemia (ischemia)-induced impairment of 2-deoxyglucose (2DG) uptake by rat hippocampal slices was evaluated. Since minaprine was found to possess both a stimulating effect on acetylcholine release and a blocking effect on 5-HT2 receptors, the improving effect of minaprine on impaired 2DG uptake was compared to the findings obtained with oxotremorine, ketanserin and pentobarbital. Hippocampal slices were exposed to 20-min ischemia, and then these slices were returned to oxygenated and glucose-containing buffer for 6 hr. Ischemia reduced 30 mM KCl-induced 2DG uptake by the hippocampus. Pretreatment with minaprine, oxotremorine, pentobarbital and ketanserin attenuated the ischemia-induced decline of 2DG uptake. In addition, minaprine, oxotremorine and pentobarbital relatively recovered the increase of 2DG uptake in the hippocampal slices under hypoxia for 45 min. The present results suggest that minaprine exerts a neuroprotective action against ischemia-induced deficit of energy metabolism in vitro.

    DOI

  • AN INVITRO CIRCADIAN-RHYTHM OF PROTEIN-SYNTHESIS IN THE RAT SUPRACHIASMATIC NUCLEUS UNDER TISSUE-CULTURE CONDITIONS

    S SHIBATA, T HAMADA, K TOMINAGA, S WATANABE

    BRAIN RESEARCH   584 ( 1-2 ) 251 - 256  1992.07

     View Summary

    Because inhibitors of protein synthesis produce changes in the circadian rhythm of wheel-running activity in rodents, we examined the circadian changes of in vitro protein synthesis by the rat suprachiasmatic nucleus (SCN). We demonstrated a robust circadian rhythm [C-14]leucine incorporation as well as [H-3]2-deoxyglucose (2DG) uptake by the SCN. The peak time of 2DG uptake was around circadian time 6-9 h (CT6-CT9). In contrast to 2DG uptake, the maximum rate of leucine incorporation occurred around CT22-CT0. Thus, the leucine incorporation rate preceded 2DG uptake by 6-9 h. Leucine incorporation was-inhibited by protein synthesis inhibitors, but not by tetrodotoxin. Since a robust circadian rhythm of leucine incorporation by the SCN was detected in vitro by using our method, this procedure may be useful to study the circadian clock function of the SCN.

  • PHASE-RESETTING EFFECT OF 8-OH-DPAT, A SEROTONIN(1A) RECEPTOR AGONIST, ON THE CIRCADIAN-RHYTHM OF FIRING RATE IN THE RAT SUPRACHIASMATIC NUCLEI INVITRO

    S SHIBATA, A TSUNEYOSHI, T HAMADA, K TOMINAGA, S WATANABE

    BRAIN RESEARCH   582 ( 2 ) 353 - 356  1992.06

     View Summary

    The 5-HTergic neurons in the mesencephalic raphe nuclei provide a robust projection to the hypothalamic suprachiasmatic nucleus (SCN), the site of a putative neuronal circadian pacemaker. Although it has been suggested that 5-HT neurons may play a role in the circadian timing system, this role has not yet been specified. Prosser et al. (Brain Res., 534 (1990) 336-339) reported that 1 h treatments with quipazine induce robust phase shifts in vitro, and that this effect depends upon the circadian time of treatment. However, quipazine is a non-specific 5-HT agonist. Besides, it is reported that the 5-HT1A agonist, 8-hydroxy-2-(di-n-propylamino)tetraline hydrobromide (8-OH-DPAT) affected a circadian rhythm of hamster wheel-running activity. In the present study we investigated whether the 5-HT1A agonist 8-OH-DPAT can reset the phase of the SCN clock when it is isolated in vitro. The present results show that 1 h treatments with 8-OH-DPAT induce robust phase advances in vitro when it was administered during the subjective day. This result suggests that 5HT1A receptor functioning may play a role in modulating the phase of SCN clock, especially during the subjective day.

    DOI

  • EFFECTS OF INHIBITORY AND EXCITATORY DRUGS ON THE METABOLIC RHYTHM OF THE HAMSTER SUPRACHIASMATIC NUCLEUS INVITRO

    K TOMINAGA, S SHIBATA, S UEKI, S WATANABE

    EUROPEAN JOURNAL OF PHARMACOLOGY   217 ( 1 ) 79 - 84  1992.06

     View Summary

    In order to elucidate the role of excitatory and inhibitory transmitters within the suprachiasmatic nucleus (SCN) in the circadian change of 2-deoxyglucose (2-DG) uptake in this nucleus, the effects of 8-hydroxy-2-(di-n-propylamino) tetralin hydrobromide (8-OH-DPAT), muscimol, flurazepam, pentobarbital and glutamate on uptake of 2-DG by hamster SCN were examined in hypothalamic slice preparations. 2-DG uptake in the SCN was high during the subjective day and low during the subjective night. The high uptake of 2-DG in the SCN during the daytime was inhibited by the superfusion of 8-OH-DPAT, muscimol, flurazepam and pentobarbital in a dose-dependent manner, but the low uptake of 2-DG during the night was unaffected. The low uptake during the night was significantly increased by treatment with glutamate, whereas 2-DG uptake during the day was unaffected. In contrast to the above results, 20 mM KCl and 1-mu-M tetrodotoxin increased and decreased 2-DG uptake during both the day and night, respectively. The present results strongly suggest that agonists of 5-HT1A receptors and GABA(A)-benzodiazepine-barbiturate complex receptors regulate the function of the SCN through their inhibitory action on 2-DG uptake during the day, and that glutamate also regulates SCN function through it stimulatory action on 2-DG uptake during the night.

    DOI

  • MORPHOLOGICAL AND FUNCTIONAL-DEVELOPMENT OF THE SUPRACHIASMATIC NUCLEUS IN TRANSPLANTED FETAL HYPOTHALAMUS

    R AGUILARROBLERO, S SHIBATA, JC SPEH, R DRUCKERCOLIN, RY MOORE

    BRAIN RESEARCH   580 ( 1-2 ) 288 - 296  1992.05

     View Summary

    The development of the suprachiasmatic nucleus (SCN) in fetal rat hypothalamus transplanted to the adult brain was studied using morphological and functional methods. Anterior hypothalamic tissue was transplanted into the third ventricle, lateral ventricle or subarachnoid space of intact, adult hosts from E17 fetuses. These transplants developed the cytoarchitectonic and immunohistochemical staining characteristics of SCN, clusters of parvocellular neurons expressing vasopressin- and vasoactive intestinal polypeptide-like immunoreactivity in adjacent cellular populations, irrespective of the exact location of the transplanted tissue in the host brain. The functional status of the translants placed in the rostral third ventricle and the foramen of Monroe was analyzed and compared to host SCN using in vitro recording of neuronal firing rate and measurement of metabolism using the 2-deoxyglucose (2-DG) technique. During subjective day, neuronal firing rates and 2-DG uptake were high in discrete cell groups within the transplants which were subsequently demonstrated to exhibit the cytoarchitectonic and immunohistochemical characteristics of SCN. The firing rates and 2-DG uptake in these areas were lower during the subjective night. This pattern of activity closely resembles that of the intact SCN. In contrast, neither transplanted anterior hypothalamic area, lacking an identifiable SCN-like structure, nor posterior hypothalamic area showed day-night differences in firing rate or 2-DG uptake. These observations indicate that SCN transplanted into intact adult hosts exhibits morphological and functional differentiation nearly identical to the host and that the transplanted SCN maintains circadian function which is probably entrained to the host SCN.

    DOI

  • EXCITATORY EFFECT OF N-METHYL-D-ASPARTATE AND KAINATE RECEPTOR ON THE 2-DEOXYGLUCOSE UPTAKE IN THE RAT SUPRACHIASMATIC NUCLEUS INVITRO

    S SHIBATA, K TOMINAGA, T HAMADA, S WATANABE

    NEUROSCIENCE LETTERS   139 ( 1 ) 83 - 86  1992.05

     View Summary

    The suprachiasmatic nuclei (SCN) have been identified as a pacemaker for many circadian rhythms in mammals. The previous findings indicate that excitatory amino acid (EAA) receptors play an important role in the transmission of light information from the retina to the circadian clocks. The 2-deoxyglucose (2DG) uptake shows a robust circadian change; high uptake during subjective day and low uptake during subjective night. To determine whether EAA agonists regulate 2DG uptake in the SCN, we have measured 2DG uptake in the rat SCN in vitro. We report that, during the subjective night, glutamate, N-methyl-D-aspartate (NMDA) and kainic acid (KA) cause an increase in 2DG uptake and that NMDA- or KA-induced increase of 2DG uptake is blocked by co-treatments with competitive and non-competitive NMDA or KA receptor antagonists. EAA receptor agonist-induced increase in 2DG uptake occurs only during subjective night, at circadian times when photic phase shifting of activity occurs. Taken together, those data suggest that EAA may be an important transmitter of light information from retina to the SCN.

    DOI

  • EFFECTS OF 5-HT(1A) RECEPTOR AGONISTS ON THE CIRCADIAN-RHYTHM OF WHEEL-RUNNING ACTIVITY IN HAMSTERS

    K TOMINAGA, S SHIBATA, S UEKI, S WATANABE

    EUROPEAN JOURNAL OF PHARMACOLOGY   214 ( 1 ) 79 - 84  1992.04

     View Summary

    The effects of 5-HT1A receptor agonists 8-hydroxy-2-(di-n-propylamino)tetralin (8-OH-DPAT), buspirone and ipsapirone on wheel-running activity in hamsters were investigated in comparison with those of GABA(A) receptor agonist muscimol and benzodiazepine triazolam. Intraperitoneal administration of 8-OH-DPAT, buspirone, ipsapirone, muscimol and triazolam at circadian time (CT) 8 (CT 12; onset of activity) induced a significant phase advance of wheel-running activity under constant light conditions. However, administration of these drugs at other CT points did not induce phase changes. The administration of trifluoromethylphenylpiperazine (TFMPP), a 5-HT1B receptor agonist. at CT8 produced a small phase advance. The phase advance induced by 8-OH-DPAT was blocked by pretreatment with (-)-pindolol, a 5-HT1A receptor antagonist. In addition, 8-OH-DPAT, buspirone and SM3997 accelerated the rate of re-entrainment to an 8-h phase advance in the light-dark cycle. These observations suggest that 5-HT1A receptors in the brain participate in the regulation of the circadian rhythm of wheel-running activity in hamsters.

    DOI

  • NEUROPROTECTIVE EFFECT OF WEB-1881-FU (NEBRACETAM) ON AN ISCHEMIA-INDUCED DEFICIT OF GLUCOSE-UPTAKE IN RAT HIPPOCAMPAL AND CEREBRAL CORTICAL SLICES AND CA1 FIELD POTENTIAL IN HIPPOCAMPAL SLICES

    S SHIBATA, Y KAGAMIISHI, S UEKI, S WATANABE

    JAPANESE JOURNAL OF PHARMACOLOGY   58 ( 3 ) 243 - 250  1992.03

     View Summary

    Effect of WEB 1881 FU (nebracetam) on hypoxia and ischemia-induced impairment of 2-deoxyglucose (2DG) uptake and CA1 field potentials induced by hypoxia and hypoxia/hypoglycemia (ischemia) in rat brain slices was evaluated and compared to the findings obtained with pentobarbital and idebenone. Hippocampal and cortical slices were exposed to 15-20 min of ischemia, and then these slices were returned to oxygenated and glucose-containing buffer for 6 hr. Ischemia reduced both 30 mM KCl-induced 2DG uptake and CA1 field potentials elicited by the stimulation of Schaffer collaterals in the hippocampus. Pretreatment of nebracetam at 1 mM or pentobarbital at 0.1 mM attenuated a decline of 2DG uptake and CA1 field potentials under the condition of ischemia. In addition, nebracetam and pentobarbital relatively recovered the increase of 2DG uptake in the hippocampus under hypoxia for 45 min. Furthermore, these drugs also attenuated the decline of 2DG uptake induced by 10 mM glutamate for 20 min. However, treatment with idebenone did not recover the deficit of 2DG uptake and CA1 field potential. The present result suggests that nebracetam and pentobarbital exert neuroprotective actions against not only ischemia but also glutamate toxicity.

    DOI

  • Brain neuronal mechanisms of circadian rhythms in mammalians

    S. Shibata, K. Tominaga

    Yakugaku Zasshi   111 ( 6 ) 270 - 283  1991

    DOI PubMed

  • 生物のサーカディアンリズム:高等動物

    柴田重信, 富永恵子

    化学と生物   29   124 - 131  1991

    DOI

  • Behavioral pharmacological and electroencephalographic effects of the 5-HT(1A) partial agonist ipsapirone

    T. Yamamoto, S. Shibata, K. Teshima, Y. Inoue, M. Ushio, K. Toninaga, M. Ohno, S. Watanabe, S. Ueki

    Folia Pharmacologica Japonica   98 ( 1 ) 41 - 52  1991  [Refereed]

    DOI PubMed

  • In vitro視交叉上核の電気活動およびdeoxyglucose uptake代謝活性リズムについて、神経研究の進歩、34、19-28、1990.

    柴田重信, 富永恵子

    神経研究の進歩   34   19 - 28  1990

    DOI

  • Effects of benzodiazepine and GABA antagonists on anticonflict effects of antianxiety drugs injected into the rat amygdala in a water-lick suppression test

    Shigenobu Shibata, Kimihiro Yamashita, Etsuko Yamamoto, Tohru Ozaki, Showa Ueki

    Psychopharmacology   98 ( 1 ) 38 - 44  1989.05  [Refereed]

     View Summary

    In order to elucidate the role of the amygdala in rat conflict behavior in a water lick suppression test, we examined the effect of lesions of various nuclei of the amygdaloid complex on this behavior. An anticonflict effect was produced by a lesion of the anterior part of central and basolateral amygdala, and lesion to the posterior part of the central amygdala, but not by posterior of the basolateral amygdala or medial amygdala lesions. These results suggest that the amygdala, especially the anterior part of the central and basolateral nuclei, plays an important role in conflict behavior of rats in the water lick test. In a second experiment, the effects of benzodiazepine- and GABA-antagonists on the anticonflict action of diazepam, zopiclone, and phenobarbital injected into the anterior part of central and basolateral amygdala were examined, also using a water lick suppression test. A dose-dependent anticonflict action was produced by systemic administration as well as by intra-amygdala injection of diazepam, zopiclone, lormetazepam, flurazepam and phenobarbital. The order of potency was lormetazepam&gt
    zopiclone≧diazepam&gt
    flurazepam ≧phenobarbital for both routes of injection. The antiamygdala effects of diazepam and zopiclone injected into the amygdala were completely reversed by Ro15-1788 and β-CCM but not by bicuculline, while the anticonflict effect of phenobarbital was reversed by β-CCM but not by Ro15-1788 or bicuculline. The present results strongly suggest that the anterior nuclei of central and basolateral amygdala are important sites of action of antianxiety drugs, and that an anticonflict action produced by intra-amygdala injection of benzodiazepines or barbiturate is mediated through the different receptor mechansims. © 1989 Springer-Verlag.

    DOI PubMed

  • Effects of melatonin on neuronal activity in the rat suprachiasmatic nucleus in vitro

    Shigenobu Shibata, Vincent M. Cassone, Robert Y. Moore

    Neuroscience Letters   97 ( 1-2 ) 140 - 144  1989.02  [Refereed]

     View Summary

    The pineal hormone melatonin is believed to act directly on the hypothalamic suprachiasmatic nuclei (SCN), which are principal circadian pacemakers in rodents, although direct demonstration of this is lacking. To determine whether the SCN are sensitive to melatonin electrophysiologically, we investigated the effect of melatonin on rat SCN single-unit activity in an in vitro hypothalamic slice preparation. SCN single-unit activity was inhibited by melatonin superfusion during the late subjective day in a dose-dependent manner but not at other times of day, supporting the view that melatonin acts directly on SCN neurons. © 1989.

    DOI PubMed

  • 選択的セロトニン取りこみ阻害作用を有する新しい抗うつ薬Paroxetineの行動薬理学的研究

    山本経之, 柴田重信, 島添隆雄, 岩崎克典, 大野益男, 皆本義基, 古谷嘉章, 宮本千葉, 渡辺繁紀, 植木昭和

    日本薬理学雑誌   94   189 - 206  1989  [Refereed]

  • Development of a fetal circadian rhythm after disruption of the maternal circadian system

    Shigenobu Shibata, Robert Y. Moore

    Developmental Brain Research   41 ( 1-2 ) 313 - 317  1988.06  [Refereed]

     View Summary

    The role of maternal circadian rhythms in the development of the fetal circadian system was investigated in the rat. Pregnant females were subjected to procedures known to disrupt circadian function, ablation of the maternal suprachiasmatic nuclei (SCN) or housing in constant illumination, on gestational day 10. Circadian function was assessed in fetuses at gestational day 22 by analysis of glucose utilization in hypothalamic slices in vitro using the 2-deoxyglucose method. Fetuses from control females exhibit a robust rhythm in glucose utilization in the SCN. In contrast, the SCN of fetuses from females with SCN lesions, or housed in constant illumination, show no significant day-night difference in glucose utilization. Analysis of individual brains indicates, however, that this apparent disruption in the development of circadian rhythmicity in metabolism in the fetal SCN is due to a desynchronization of individual fetuses resulting from the loss of maternal entraining influences. Thus, the fetal SCN is capable of developing a circadian rhythm in glucose utilization independent of the maternal circadian system. © 1988.

    DOI

  • Involvement of the medial amygdaloid nucleus in the action of imipramine in rats subjected to the forced swimming test

    Takao Shimazoe, Shigenobu Shibata, Shin-Ichi Yatsugi, Showa Ueki

    journal of pharmacobio-dynamics   11 ( 2 ) 137 - 139  1988.02  [Refereed]

     View Summary

    A forced swimming test proposed by Porsolt1) is a useful method for screening antidepressants. To evaluate drug effects more objectively, vibration of the walls of a water tank caused by the escape behavior of the rat was recorded. Imipramine (IMI) increased the number of vibration and this effect was observed in the medial amygdala lesioned rat but not in the central or basolateral amygdala le-sioned rat. The present result suggests that the medial amygdala is an important site of action of IMI. © 1988, The Pharmaceutical Society of Japan. All rights reserved.

    DOI PubMed

  • Electrical and metabolic activity of suprachiasmatic nucleus neurons in hamster hypothalamic slices

    Shigenobu Shibata, Robert Y. Moore

    Brain Research   438 ( 1-2 ) 374 - 378  1988.01  [Refereed]

     View Summary

    Single unit neuronal activity and glucose utilization were studied in the suprachiasmatic nucleus (SCN) in hypothalamic slices from the golden hamster brain in vitro. An apparent circadian rhythm was observed both in SCN single unit activity and in glucose utilization. These observation indicate that the brain slice method is useful for the analysis of pacemaker function in hamster SCN. © 1988.

    DOI PubMed

  • 攻撃行動と脳内アミン

    植木昭和, 柴田重信

    代謝   25   39 - 47  1988

  • Neuropeptide Y and Vasopressin Effects on Rat Suprachiasmatic Nucleus Neurons In Vitro

    Shigenobu Shibata, Robert Y. Moore

    Journal of Biological Rhythms   3 ( 3 ) 265 - 276  1988  [Refereed]

     View Summary

    The effect of neuropeptide Y (NPY) and vasopressin (VP) on the firing rates of neurons in the retinorecipient portion of the rat suprachiasmatic nucleus (SCN) was studied by superfusion onto hypothalamic slices in vitro. NPY produces either excitation or excitation followed by inhibition when applied during subjective day. With increasing doses of NPY, the number of neurons showing an excitatory response alone remains stable, and the neurons increase firing rates slightly. In contrast, the number of neurons showing an excitatory re sponse followed by inhibition is greater with increased NPY concentration, and the inhibitory effect is prolonged. VP produces only excitatory responses, and these are increased with increasing VP concentration. Prolonged administration of NPY produces an overall decrease in firing rate during subjective day, but has no effect during subjective night. In contrast, the excitatory effects of VP are essentially the same in both subjective day and subjective night. The responses to NPY and VP are preserved in calcium-free medium, indicating that the peptides act directly on SCN neurons. Optic nerve stimulation produces excitation in 36% of neurons in the retinorecipient component of the SCN, and 80% of these respond to NPY, whereas only 29% of neurons that do not respond to optic nerve stimulation are affected by NPY. In contrast, both groups respond equally well to VP. These observations indicate that neurons in the component of the rat SCN receiving retinal afferents respond differently to NPY and VP, and that responses to NPY are complex and dependent upon the circadian time at which NPY is administered. © 1988, Sage Publications. All rights reserved.

    DOI

  • Effects of calcium ions on glucose utilization in the rat suprachiasmatic nucleus in vitro

    Shigenobu Shibata, George C. Newman, Robert Y. Moore

    Brain Research   426 ( 2 ) 332 - 338  1987.11  [Refereed]

     View Summary

    The role of calcium ions in maintenance of the circadian rhythm in glucose utilization in the suprachiasmatic nucleus (SCN) of the hypothalamus was investigated in vitro in a rat hypothalamic slice preparation using the 2-deoxyglucose (2-DG) method. In normal Krebs solution, 2-DG uptake of adult and embryonic day 22 rat SCN was higher in subjective day than in subjective night. In calcium-free Krebs solution, however, 2-DG uptake of adult and embryonic SCN was low in both subjective day and night periods. These results indicate that the SCN rhythm in metabolic activity is dependent on calcium ions in both adult and embryonic rats. Since immunohistochemical and ultrastructural analysis of synapse formation has shown very few synapses in the SCN of embryonic day 22 rats, it is suggested that the development and maintenance of the circadian rhythm in metabolism demonstrated by the 2-DG method depends on intracellular calcium-mediated events rather than synaptic transmission. © 1987.

    DOI PubMed

  • A new forced swimming test for the evaluation of antidepressants in rats by recording vibration of a water tank

    Takao Shimazoe, Shigenobu Shibata, Showa Ueki

    Journal of Pharmacobio-Dynamics   10 ( 11 ) 639 - 643  1987  [Refereed]

     View Summary

    In order to more objectively evaluate drug effects in the forced swimming test proposed by Porsolt et a I. u as a screening method for antidepressants, vibrations of the wall of a tank caused by rats trying to escape from water were recorded. Locomotor activity was also measured in an activity cage. Male Wistar rats were forced to swim once daily for 15 min in a tank of 25 °C water filled to a depth of 20 cm. After 4 d sessions of swimming, drugs were administered i.p. 3 times (24, 5 and 1 h prior to the test session). On day 5,45 min after the last injection of a test drug, locomotor activity was measured for 15 min and then the rat was subjected to the forced swimming test. All antidepressants tested, dose-dependently increased tank vibration. Nomifensine, atropine, methamphetamine, chlorpheniramine and diazepam increased locomotor activity. The remaining drugs had no effect or reduced locomotion. In addition, the pattern of the tank vibrations, caused by rats treated with most drugs, like atropine, showed a burst during the first 5 min followed by sporadic vibrations. Nomifensine and methamphetamine, on the other hand, caused vibrations throughout the 15 min test session. A specific effect of antidepressants was revealed by this forced swimming test in combination with the measurement of locomotor activity. © 1987, The Pharmaceutical Society of Japan. All rights reserved.

    DOI PubMed

  • Development of neuronal activity in the rat suprachiasmatic nucleus

    Shigenobu Shibata, Robert Y. Moore

    Developmental Brain Research   34 ( 2 ) 311 - 315  1987  [Refereed]

     View Summary

    The development of function was studied in the rat suprachiasmatic nucleus (SCN) by analyzing the rate and pattern of neuronal discharge in vitro from embryonic day 22 (E22) to postnatal day 14 (P14), and comparing these with adult SCN. The firing rate of SCN neurons on E22 is low but there is a clear circadian rhythm with subjective day values (1.6 ± 0.2 impulses/s) significantly higher than those for subjective night (0.9 ± 0.2 impulses/s). At E22 most neurons show an irregular firing pattern. The firing rate of SCN neurons gradually increases from E22 to P14 with emergence of patterns of firing of individual neurons that approximate those of adult SCN. In calcium-free medium the firing rate of SCN neurons at P1 is reduced and the circadian rhythm in firing rate is abolished. These results demonstrate that SCN neurons exhibit a circadian rhythm in firing rate early in development, largely prior to the formation of synaptic contacts within the SCN, and indicate that expression of the rhythm requires the presence of calcium ions. © 1987.

    DOI

  • Behavioral and Electroencephalographic Effects of Zopiclone, a Cyclopyrrolone Derivative

    Showa Ueki, Shigenori Watanabe, Tsuneyuki Yamamoto, Yasufumi Kataoka, Shigenobu Shibata, Kazuhiko Shibata, Hisashi Ohta, Takao Shimazoe, Hirosuke Kawamoto

    The Japanese Journal of Pharmacology   43 ( 3 ) 309 - 326  1987  [Refereed]

     View Summary

    Behavioral effects of zopiclone were investigated in mice and rats and compared with the data on diazepam, nitrazepam and flurazepam. The electro encephalographic effect of the drug was also examined in unanesthetized rabbits with chronic electrode implants and compared with that of diazepam. The present results indicate that zopiclone possesses pharmacological properties qualitatively similar to benzodiazepines, which are characterized by potent anticonflict and antiaggressive effects and much weaker anticonvulsant, muscle relaxant, ataxiogenic, sedative and anesthesia potentiating effects
    the properties of this drug were com pared with those of diazepam, nitrazepam and flurazepam. Zopiclone suppressed the EEG arousal responses and inhibited afterdischarges induced by electrical stimulation of the hippocampus and amygdala. The effects of zopiclone on EEG and afterdischarges were approximately 1/10 those of diazepam. © 1987, The Japanese Pharmacological Society. All rights reserved.

    DOI PubMed

  • Effects of dibutyryl cyclic adenosine monophosphate and dibutyryl cyclic guanosine monophosphate on neuron activity of suprachiasmatic nucleus in rat hypothalamic slice preparation

    Shyh Yuh Liou, Shigenobu Shibata, Akiko Shiratsuchi, Showa Ueki

    Neuroscience Letters   67 ( 3 ) 339 - 343  1986.06  [Refereed]

     View Summary

    In order to elucidate the role of cyclic nucleotides in the suprachiasmatic nucleus, we examined the effect of dibutyryl cyclic AMP (c-AMP) and dibutyryl cyclic GMP (c-GMP) on single neuronal activity of the suprachiasmatic nucleus of the rat. A hypothalamic slice about 300 μm thick, including the suprachiasmatic nucleus, was cut coronally using a vibratome. By means of bath- as well as iontophoretic application, c-AMP and c-GMP caused mainly an inhibition and facilitation in suprachiasmatic neuronal activity, respectively. The present results suggest that c-AMP and c-GMP play an important role in regulation of suprachiasmatic neuronal activity as a second messenger and that these two nucleotides may function as a reciprocal messenger. © 1986.

    DOI PubMed

  • Inhibitory action of insulin on suprachiasmatic nucleus neurons in rat hypothalamic slice preparations

    Shigenobu Shibata, Shyh Yuh Liou, Showa Ueki, Yutaka Oomura

    Physiology and Behavior   36 ( 1 ) 79 - 81  1986  [Refereed]

     View Summary

    In order to elucidate a role of insulin on neurons in the suprachiasmatic nucleus (SCN) involved in the generation of circadian feeding behavior, we examined the effect of insulin on SCN neuronal activity at in vitro condition. Bath application of insulin (1-100 μU) mainly inhibited the SCN neuronal activity, and this inhibitory effect was still observed in a Ca2+-free Krebs solution. The present result strongly suggests that insulin directly inhibits SCN neurons and may explain an increase in food intake by insulin infusion into the SCN during the light period. © 1986.

    DOI PubMed

  • Influence of excitatory amino acid receptor antagonists and of baclofen on synaptic transmission in the optic nerve to the suprachiasmatic nucleus in slices of rat hypothalamus

    S. Shibata, S. Y. Liou, S. Ueki

    Neuropharmacology   25 ( 4 ) 403 - 409  1986  [Refereed]

     View Summary

    Electrical stimulation of the optic nerve evoked two positive waves with short latency, followed by a large negative wave in the suprachiasmatic nucleus of slices of hypothalamus of the rat. The latency to peak of the two positive waves and the large negative wave were 2.7 ± 0.1, 6.1 ± 0.1 and 10.3 ± 0.5 msec, respectively. Only the large negative wave disappeared in low calcium Ca2+-high magnesium (Mg2+) Krebs solution and with the addition of tetrodotoxin (1 μM) all the waves disappeared. Baclofen inhibited the large negative wave in a dose-dependent manner but not the two positive waves. Excitatory amino acid antagonists also inhibited only the large negative wave, i.e. it was reduced to about 70% by 1 mM glutamic acid diethyl ester and to about 50% by both 1 mM 2-amino-4-phosphonobutyric acid and 1 mM dl-2-amino adipic acid. All waves were unaffected by 0.1 mM atropine, hexamethonium and curare. These results indicate that two positive waves, induced by stimulation of the optic nerve are attributed to nerve conduction and the large negative wave to the neurons of the supraehiasmatic nucleus, and that the neuronal pathway from the optic nerve to the suprachiasmatic nucleus may include aspartate and/or glutamate as an excitatory neurotransmitter. © 1986.

    DOI PubMed

  • Optic nerve stimulation-induced increase of release of 3h-glutamate and 3h-aspartate but not 3h-gaba from the suprachiasmatic nucleus in slices of rat hypothalamus

    Shyh Yuh Liou, Shigenobu Shibata, Katsunori Iwasaki, Showa Ueki

    Brain Research Bulletin   16 ( 4 ) 527 - 531  1986  [Refereed]

     View Summary

    This study investigates the possibility of glutamate and/or aspartate as an excitatory transmitter in the retinohypothalamic fibers' terminal within the anterior hypothalamus, including the suprachiasmatic nucleus (SCN). We studied the release of [3]H-Glu and 3H-Asp from the SCN of rat hypothalamic slices produced by optic nerve stimulation. Stimulation of the optic nerve released 3H-Glu, 3H-Asp but not 3H-GABA, while stimulation of the SCN released all three. These releases were suppressed in both Ca2+ free Krebs and tetrodotoxin (TTX) 1 μM containing Krebs. The present result with previous electrophysiological observations strongly suggest that glutamate and/or aspartate are included in this retinohypothalamic termination, while GABA is included in intrinsic and/or extrinsic neurons of the SCN, excluding the terminal of the retinohypothalamic fibers. © 1986.

    DOI PubMed

  • Effect of monoamines on field potentials in the suprachiasmatic nucleus of slices of hypothalamus of the rat evoked by stimulation of the optic nerve

    S. Y. Liou, S. Shibata, S. Ueki

    Neuropharmacology   25 ( 9 ) 1009 - 1014  1986  [Refereed]

     View Summary

    The effects of the application of serotonin, histamine, noradrenaline and dopamine to the bath on field potentials in the suprachiasmatic nucleus evoked by stimulation of the optic nerve were studied using a hypothalamic slice. Stimulation of the contralateral optic nerve evoked fast positive and late large negative waves in the suprachiasmatic nucleus. The monoamines produced a dose dependent suppression of the amplitude of the negative wave but did not affect that of the positive waves, and the order of potency was serotonin &gt
    noradrenaline &gt
    dopamine ≥ histamine. The negative wave was suppressed by phenylephrine (0.1-10 μ M) in a dose-dependent manner, whereas it was unaffected by isoproterenol (0.1-10 μM). The suppression of the negative wave produced by the application of histamine and noradrenaline was antagonized by the H1-receptor antagonist, diphenhydramine and the α1-receptor antagonists, phenoxybenzamine and phentolamine. Therefore, the suppression of the negative wave by histamine and noradrenaline was mediated by the H1-receptor and α1-receptor, respectively. The present study indicates that monoamines may play an inhibitory role in the regulation of neurotransmission in the retinohypothalamic pathway to the suprachiasmatic nucleus. © 1986.

    DOI PubMed

  • Changes in brain catecholamine levels following olfactory bulbectomy and the effect of acute and chronic administration of desipramine in rats

    Katsunori Iwasaki, Michihiro Fujiwara, Shigenobu Shibata, Showa Ueki

    Pharmacology, Biochemistry and Behavior   24 ( 6 ) 1715 - 1719  1986  [Refereed]

     View Summary

    Bilateral olfactory bulbectomy of the rat caused marked changes of noradrenaline level in several brain regions accompanied with the development of mouse-killing behavior (muricide). Noradrenaline level increased in the medial amygdala, ventromedial and lateral hypothalamus in muricidal olfactory bulbectomized rats (OB rats) but not in non-muricidal OB rats, while dopamine level decreased in the lateral hypothalamus in muricidal OB rats. Acute administrations of desipramine not only suppressed muricide of OB rats but normalized noradrenaline change in ventromedial hypothalamus and dopamine change in lateral hypothalamus. Chronic administration of desipramine also suppressed muricide and normalized noradrenaline changes in ventromedial and lateral hypothalamus and medial amygdala. These findings suggest that the increase in noradrenaline levels in the medial amygdal, ventromedial and lateral hypothalamus may be important for the induction of muricide in OB rats, and muricide was suppressed by desipramine in accordance with the normalization of increased noradrenaline levels, and that the change in dopaminergic function in the lateral hypothalamus may also be important for this muricide. © 1986.

    DOI PubMed

  • 視交叉上核ニューロンの機能分化

    大村裕, 柴田重信

    神経研究の進歩   29   58 - 70  1985

    DOI

  • Behavioral and electroencephalographic effects of lormetazepam

    Showa Ueki, Shigenori Watanabe, Michihiro Fujiwara, Tsuneyuki Yamamoto, Shigenobu Shibata, Kazuhiko Shibata, Hisashi Ohta, Yasuko Sakurai, Katsunori Iwasaki, Shyh Yuh Liou, Kimihiro Yamashita, Takao Shimazoe, Takayuki Yamaji

    Folia Pharmacologica Japonica   86 ( 2 ) 145 - 163  1985  [Refereed]

     View Summary

    Behavioral and electroencephalographic effects of lormetazepam were investigated in rats, mice and rabbits, in comparison with those of diazepam, nitrazepam and flurazepam. Locomotor activity of mice in an open-field situation was decreased with large doses of lormetazepam and diazepam, while it was increased with nitrazepam and flurazepam. The anticonflict effect of lormetazepam in rats was much more potent than those of diazepam, nitrazepam and flurazepam. In suppressing the muricide of olfactory bulbectomized and raphe lesioned rats, lormetazepam was more potent than diazepam, nitrazepam and flurazepam. Lormetazepam, diazepam and flurazepam prevented both maximal electroshock and pentetrazol convulsions in mice, the effects on the latter being much more potent than those on the former. Lormetazepam was more potent than diazepam and flurazepam in potentiating thiopental-, ether- and ethanol anesthesia, impairing rotarod performance and in muscle relaxant activity. In conscious rabbits with chronically implanted electrodes, lormetazepam induced a drowsy EEG pattern and suppressed the EEG arousal responses not only to auditory stimulation but also to electrical stimulation of the midbrain reticular formation or hypothalamus. Lormetazepam also inhibited afterdischarges induced by electrical stimulation of the hippocampus and amygdala. These results indicate that lormetazepam has pharmacological properties characteristic of benzodiazepines and that the activity is more potent than those of diazepam, nitrazepam and flurazepam. © 1985, The Japanese Pharmacological Society. All rights reserved.

    DOI PubMed

  • Behavioral pharmacology of amantadine with special references to the effect on abnormal behavior in mice and rats

    Michihiro Fujiwara, Yasuko Sakurai, Yoshihiro Kiyota, Takao Shimazoe, Ohta Hisashi, Shigenobu Shibata, Showa Ueki

    Folia Pharmacologica Japonica   85 ( 4 ) 259 - 274  1985  [Refereed]

     View Summary

    Behavioral effects of amantadine, especially on abnormal behavior in mice and rats, were reevaluated, in comparison with those of tricyclic antidepressants, methamphetamine and L-DOPA. 1) Amantadine at 10~50 mg/kg, i.p., tended to decrease ambulation and rearing in rats and mice. The drug at 50 mg/kg, i.p., caused piloerection and hyperirritability and at doses over 80 mg/kg, i.p., it impaired coordinated motor activity in rats. 2) Amantadine inhibited methamphetamine-induced hyperactivity, but apomorphine-induced stereotyped behavior was unaffected in rats. 3) Amantadine was equipotent to imipramine in suppressing haloperidol-induced catalepsy in rats, but L-DOPA was without effect. On the other hand, amantadine was 40 and 400 times as potent as imipramine and L-DOPA, respectively, in suppressing Δ9-tetrahydrocannabinol (THG)-induced catalepsy in rats. 4) Amantadine was as potent as imipramine in suppressing the muricide of olfactory bulbectomized rats, but was 3.5, 8.8 and 225.5 times as potent as methamphetamine, imipramine and L-DOPA, respectively, in inhibiting THC-induced reversible muricide in rats. 5) Amantadine at 50 mg/kg did not elicit circling behavior in the rat with unilateral nigral lesion induced by 6-hydroxydopamine. 6) Amantadine at high doses caused irritable aggression characterized by squealing in rats pretreated with intraventricular 6-hydroxydopamine. The most important characteristic of amantadine is its prominent effect suppressing the THC-induced catalepsy and muricide. This may be a reflection of the feature of amantadine activating the dopaminergic as well as the serotonergic systems. © 1985, The Japanese Pharmacological Society. All rights reserved.

    DOI PubMed

  • Differential Effects of Electroconvulsive Shock on Four Models of Experimentally-Induced Aggression in Rats

    Shyh Yuh Liou, Shigenobu Shibata, Showa Ueki

    The Japanese Journal of Pharmacology   37 ( 2 ) 167 - 172  1985  [Refereed]

     View Summary

    The effects of electroconvulsive shock (ECS) on the hyperemotionality and muricide in olfactory bulbectonnized rats (OB rat) were compared with those in spontaneous killer rats (SP rat), raphe lesioned rats (Raphe rat) and z9-tetra-hydrocannabinol treated rats (THC rat). Single and chronic treatment of ECS inhibited the muricide and attack response to a rod, but did not affect the struggle response to handling and flight response to air blowing. Muricide was markedly inhibited by ECS in OB and SP rats, and was moderately inhibited in THC rats, while it was slightly inhibited in Raphe rats. The present result indicates that muricide of OB rats is a useful model for evaluating antidepressant drugs as this behavior is markedly inhibited by ECS in OB rats. Furthermore, it is suggested that the effect of ECS on muricide is different depending upon the methods to induce muricide, although muricide itself seems to be behaviorally similar. © 1985, The Japanese Pharmacological Society. All rights reserved.

    DOI PubMed

  • The role of central noradrenergic neurons in electroconvulsive shock-induced muricide inhibition in olfactory bulbectomized rats

    Liou Shyh Yuh, Shibata Shigenobu, Ueki Showa

    Pharmacology, Biochemistry and Behavior   23 ( 1 ) 65 - 70  1985  [Refereed]

     View Summary

    In order to elucidate the role of central monoaminergic neurons in electroconvulsive shock (ECS)-induced muricide inhibition in the olfactory bulbectomized rat (OB rat), we examined the effect of chemical or electrical lesions of each monoamine-containing neuron on ECS-induced muricide inhibition. ECS-induced muricide inhibition was antagonized by 6-hydroxydopamine (6-OHDA) treatment and locus coeruleus lesions, while it was unaffected by desipramine + 6-OHDA, substantia nigra lesion, and desipramine + 5,7-dihydroxytryptamine. The present results strongly suggest that ECS-induced muricide inhibition of the OB rat is due to increased activity of the noradrenaline but not of dopamine and serotonin neurons. © 1985.

    DOI PubMed

  • The role of calcium ions in circadian rhythm of suprachiasmatic nucleus neuron activity in rat hypothalamic slices

    Shigenobu Shibata, Akiko Shiratsuchi, Shyh Yuh Liou, Showa Ueki

    Neuroscience Letters   52 ( 1-2 ) 181 - 184  1984.11  [Refereed]

     View Summary

    The role of calcium ions in maintaining the circadian rhythm of suprachiasmatic nucleus (SCN) neuron activity was investigated using rat hypothalamic slice preparations. In normal Krebs solution, the firing rate of SCN neurons was higher in the light period than in the dark period. In Ca2+ -free Krebs solution, SCN neuron activity was low during all periods and did not show diurnal rhythm. These results suggest that the disappearance of circadian rhythmic change of SCN neuron activity in Ca2+ -free Krebs solution may be due to the disappearance of synaptic transmission in the SCN. © 1984.

    DOI PubMed

  • Influence of environmental light-dark cycle and enucleation on activity of suprachiasmatic neurons in slice preparations

    Shigenobu Shibata, Yuh Liou Shyh, Showa Ueki, Yutaka Oomura

    Brain Research   302 ( 1 ) 75 - 81  1984.06  [Refereed]

     View Summary

    The influence of environmental light-dark cycle (LD) and bilateral enucleation on single neuronal activity in the suprachiasmatic nucleus (SCN) was examined using a hypothalamic slice preparation. Firstly, we reconfirmed previous results that the discharge rate in slices taken from animals kept on normal LD was higher during the light than during the dark period. Secondly, the day time discharge rate in the ventrolateral part of the SCN was decreased by bilateral enucleation and DD housing, while in the dorsomedial part it was unaffected. Thirdly, LL housing suppressed the discharge rates in both parts during the day and night periods. The present results suggest that the dorsomedial part of the SCN is more important in regulation of the circadian rhythm of SCN neuronal activity than the ventrolateral. © 1984.

    DOI PubMed

  • Responses of suprachiasmatic nucleus neurons to optic nerve stimulation in rat hypothalamic slice preparation

    Shigenobu Shibata, Yutaka Oomura, Kiichi Hattori, Hitoshi Kita

    Brain Research   302 ( 1 ) 83 - 89  1984.06  [Refereed]

     View Summary

    Stimulation of the optic nerve evoked two positive waves (P1 and P2) with short latencies and a following large negative wave (N) in the suprachiasmatic nucleus (SCN) of hypothalamic slice preparations. The conduction velocities of the P1, P2 and N waves were assumed to be 2.9 m/s, 0.5 m/s and 0.4 m/s, respectively. The N wave disappeared in low Ca2+ medium and all responses disappeared when TTX was added to the medium at a concentration of 10-6 M. The results indicate that the P1 and P2 waves were responses of the optic nerve in the SCN and the N wave was that of SCN neurons. Following optic nerve stimulation 15 out of 212 single SCN neurons were excited
    10 were excited then inhibited, 14 were excited followed by rhythmic oscillation, and 4 were inhibited. The results demonstrate neuronal projections from the optic nerve to the SCN, and suggest that the major role of the excitatory influence on SCN neurons is responsible for the elevation of neuronal activity in the SCN during the daytime. © 1984.

    DOI PubMed

  • Effects of Adrenergic Blockers on the Inhibition of Muricide by Chronic Administration of Desipramine in Oflactory Bulbectomized Rats

    Shigenobu Shibata, Hiroshi Nakanishi, Showa Ueki

    Japanese Journal of Pharmacology   35 ( 1 ) 73 - 75  1984  [Refereed]

    DOI PubMed

  • Behavioral Effects of Brotizolam, a New Thienotriazolodiazepine Derivative

    Showa Ueki, Shigenori Watanabe, Tsuneyuki Yamamoto, Shigenobu Shibata, Kazuhiko Shibata

    The Japanese Journal of Pharmacology   35 ( 3 ) 287 - 299  1984  [Refereed]

     View Summary

    The behavioral effect of brotizolam was investigated in nnice and rats, in comparison with those of diazepam, nitrazepam and estazolam. Locomotor activity of rats in an open field situation was slightly increased with smaller doses of brotizolam and estazolam and with larger doses of nitrazepam, while it was decreased with large doses of brotizolam and estazolam. The anticonflict effect of brotizolam in rats was approximately as potent as that of diazepam and was augmented following chronic administration for 10 days. In suppressing hyperemotionality and muricide of olfactory bulbectomized rats, brotizolam was more potent than diazepam, being approximately equipotent to nitrazepam and estazolam. Brotizolam, diazepam, nitrazepam and estazolam prevented both maximal electroshock and pentetrazol convulsions in mice, the effects on the latter being much more potent than those on the former. In impairing rotarod performance, brotizolam was as potent as estazolam and nitrazepam and was much more potent than diazepam in mice, but was less potent than estazolam and nitrazepam in rats. These results indicate that brotizolam possesses pharmacological properties characteristic to benzodiazepines and that the activity is more potent than that of diazepam and approximately as potent as those of nitrazepam and estazolam. © 1984, The Japanese Pharmacological Society. All rights reserved.

    DOI PubMed

  • Effects of chronic administration of antidepressants on mouse-killing behavior (muricide) in olfactory bulbectomized rats

    Shigenobu Shibata, Hiroshi Nakanishi, Shigenori Watanabe, Showa Ueki

    Pharmacology, Biochemistry and Behavior   21 ( 2 ) 225 - 230  1984  [Refereed]

     View Summary

    Two forms of drug administration, i.e., systemic subcutaneous administration and microinjection into the medial amygdala were employed to examine the effect of chronic administration of psychotropic drug on muricide in olfactory bulbectomized rats. Muricide inhibition induced by the systemic doses of chlorpromazine (CPZ) 10 mg/kg and diazepam 10 mg/kg was reduced with chronic administration, while that by desipramine (DMI) 10 mg/kg and amitriptyline 30 mg/kg was augmented with chronic adminstration. Muricide inhibition induced by microinjection of CPZ was also reduced, while that by DMI was augmented. These results indicate that muricide by olfactory bulbectomized rats is a useful animal model for evaluating antidepressants and that a potential site of action of antidepressants is located in the medial amygdala. © 1984.

    DOI PubMed

  • Electrophysiological studies of the development of suprachiasmatic neuronal activity in hypothalamic slice preparations

    Shigenobu Shibata, Yutaka Oomura, Shyh Yuh Liou, Showa Ueki

    Developmental Brain Research   13 ( 1 ) 29 - 35  1984  [Refereed]

     View Summary

    The rate and pattern of neuronal discharge in the suprachiasmatic nucleus (SCN) during developmental stages were studied and compared with those of the ventromedial (VMH) and the lateral hypothalamus (LHA) using rat hypothalamic slices. The firing rate of the SCN neurons was low on the 7th and 11th days
    however, it dramatically increased by the 14th day to reach the adult rate, while firing rates of VMH and LHA neurons increased gradually with age. The discharge rate of neurons in the ventrolateral part of the SCN (VL-SCN) was higher than that of the dorsomedial SCN (DM-SCN) neurons in 14-, 21- and 70-100-day-old rats. Activity of the DM-SCN neurons on day 21 were unaffected by bilateral enucleation on the third day, while activity in the VL-SCN decreased
    that of both parts was significantly decreased by a constant light schedule. © 1984.

    DOI

  • Development of the circadian rhythm of neuronal activity in suprachiasmatic nucleus of rat hypothalamic slices

    Shigenobu Shibata, Shyh Yuh Liou, Showa Ueki

    Neuroscience Letters   43 ( 2-3 ) 231 - 234  1983.12  [Refereed]

     View Summary

    Development of the circadian rhythm of neuronal activity in the suprachiasmatic nucleus (SCN) was studied in rat hypothalamic slices. The firing rate of SCN neurons of 7- and 11-day-old rat pups was low during all of the day, whereas that of 14- and 21-day-old pups was high during daytime and low during nighttime. The present results suggest that the circadian rhythm of SCN neuronal activity is established between 11 and 14 days of age, corresponding closely to the time of onset of the other various hormonal and behavioral circadian rhythms. © 1983.

    DOI PubMed

  • Inhibitory and excitatory effects of histamine on suprachiasmatic neurons in rat hypothalamic slice preparation

    Shyh Yuh Liou, Shigenobu Shibata, Katsunori Yamakawa, Showa Ueki

    Neuroscience Letters   41 ( 1-2 ) 109 - 113  1983.10  [Refereed]

     View Summary

    The effects of histamine (HA) and serotonin (5-HT) on single neurons of the suprachiasmatic nucleus (SCN) in hypothalamic slices were investigated by iontophoretic application. Sixty-two of 181 cells were inhibited by HA, whereas 30 were excited. Forty-nine of 163 cells were inhibited by 5-HT, whereas 9 were excited. Pyrilamine, an H1-antagonist, slightly (2/7), and cimetidine, an H2-antagonist, strongly (10/13) antagonized the inhibitory effect of HA. The excitatory effect of HA was not antagonized by cimetidine (0/4), but was perfectly antagonized by pyrilamine (5/5). The present results suggest that HA and 5-HT play mainly an inhibitory role in the SCN neuronal activity, and that inhibition and excitation induced by HA may be mediated by H2- and H1-receptors, respectively. © 1983.

    DOI PubMed

  • Different effects of amino acids, acetylcholine and monoamines on neuronal activity of suprachiasmatic nucleus in rat pups and adults

    Shigenobu Shibata, Shyh Yuh Liou, Showa Ueki

    Neuroscience Letters   39 ( 2 ) 187 - 192  1983.08  [Refereed]

     View Summary

    A study was undertaken of the effects of iontophoretically applied amino acids, acetylcholine and monoamines on suprachiasmatic nucleus (SCN) neurons of rat pups, at the 11th postnatal day, using a hypothalamic slice preparation. GABA and taurine inhibited 97 and 42% of SCN neurons, respectively, while glycine had no effect. Glutamate excited 68% of them. These effects were similar to those in adult rats. Serotonin and noradrenaline inhibited 6 and 10% of SCN neurons, respectively, in rat pups, whereas 32 and 26% of them were inhibited in adult rats. Acetylcholine excited 7% but inhibited 26% of SCN neurons in rat pups and these values were almost the same as those of adult rats. These results reveal that acetylcholine and amino acids regulate SCN neurons even in rat pups, whereas monoamines play an important regulatory role in SCN activity in adult rats only. © 1983.

    DOI PubMed

  • Behavioral effects of ethyl loflazepate and its metabolites

    Showa Ueki, Shigenori Watanabe, Tsuneyuki Yamamoto, Shigenobu Shibata, Kazuhiko Shibata, Hisashi Ohta, Katsuko Ikeda, Yoshihiro Kiyota, Yoko Sato

    Folia Pharmacologica Japonica   82 ( 5 ) 395 - 409  1983  [Refereed]

     View Summary

    The behavioral effects of ethyl loflazepate and its metabolites were investigated in mice and rats, and they were compared with those of diazepam, nitrazepam and lorazepam. Locomotor activity of rats in open-field situation was increased with a wide range of doses of ethyl loflazepate and with large doses of nitrazepam. The anticonflict effect of ethyl loflazepate was slightly more potent than that of diazepam and much more potent than that of lorazepam. In suppressing muricide of both olfactory bulbectomized and raphe lesioned rats, ethyl loflazepate was approximately as potent as diazepam and much less potent than nitrazepam and lorazepam. Ethyl loflazepate was more potent than diazepam, nitrazepam and lorazepam in preventing pentetrazol-induced convulsion. In potentiating thiopental anesthesia in mice and impairing rotarod performance in mice and rats, ethyl loflazepate was less potent than diazepam, nitrazepam and lorazepam. The pharmacological activities of CM6913 and CM7116, metabolites of ethyl loflazepate, were approximately as potent as that of ethyl loflazepate. The duration of action of ethyl loflazepate was longer than those of the metabolites. These results indicate that ethyl loflazepate possesses pharmacological properties characteristic to benzodiazepines, and it is approximately equal to diazepam in potency and has a longer duration of action than diazepam, nitrazepam and lorazepam. © 1983, The Japanese Pharmacological Society. All rights reserved.

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  • Effect Of Amino Acids and Monoamines on the Neuronal Activity of Suprachiasmatic Nucleus in Hypothalamic Slice Preparations

    Shigenobu Shibata, Shyh Yuh Llou, Showa Ueki

    The Japanese Journal of Pharmacology   33 ( 6 ) 1225 - 1231  1983  [Refereed]

     View Summary

    Influences of amino acids and monoamines on the single unit discharges of the suprachiasmatic nucleus (SCN) were investigated using hypothalamic slice preparations. lontophoretic application of GABA inhibited 90% and taurine inhibited 40% of SCN neurons, while glycine inhibited only 4%. L-glutamate excited about 50% of the neurons. Serotonin, noradrenaline and dopamine inhibited 30,22 and 26% of the SCN neurons, respectively. Amino acid effects were observed equally in both the ventrolateral part of the SCN and the remaining part of it, while monoamine effects were observed preferentially in the ventrolateral part of the SCN where the optic fibers and serotonin nerves terminated. These results suggest that amino acids and monoamines modulate the neuronal activity of the SCN through their direct effects. © 1983, The Japanese Pharmacological Society. All rights reserved.

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  • Circadian rhythmic changes of neuronal activity in the suprachiasmatic nucleus of the rat hypothalamic slice

    Shigenobu Shibata, Yutaka Oomura, Hitoshi Kita, Kiichi Hattori

    Brain Research   247 ( 1 ) 154 - 158  1982.09  [Refereed]

     View Summary

    The present study, utilizing rat brain slice of the suprachiasmatic nuclei (SCN) demonstrated single neuronal activity of SCN cells which changed in correspondence to environmental light-dark (L-D) schedules. The unit discharge rates of cells from animals kept on normal LL-D) cycles reached peaks at about 14.00 h and had a lowest point at around 02.00 h. The unit activity in slices from animals kept on reversed (L-D) cycles was shifted approximately 12 h. © 1982.

    DOI PubMed

  • Excitatory effects of the suprachiasmatic nucleus on the ventromedial nucleus in the rat hypothalamic slice

    Hitoshi Kita, Shigenobu Shibata, Yutaka Oomura, Kosuke Ohki

    Brain Research   235 ( 1 ) 137 - 141  1982.03  [Refereed]

    DOI PubMed

  • 視交叉上核と食欲中枢との神経生理学的関係

    大村裕, 柴田重信

    蛋白質核酸酵素   27   391 - 403  1982

  • Neuronal control of the ventromedial nucleus from the periventricular nucleus in the hypothalamic slice of the rat

    Shigenobu Shibata, Yutaka Oomura, Nobuaki Hori

    Brain Research Bulletin   8 ( 6 ) 613 - 616  1982  [Refereed]

     View Summary

    SHIBATA, S., Y. OOMURA AND N. HORI. Neuronal control of the ventromedial nucleus from the periventricular nucleus in the hypothalamic slice of the rat. BRAIN RES. BULL. 8(6) 613-616, 1982.-A neuronal pathway between periventricular nucleus and ventromedial hypothalamus (VMH) was investigated by both anatomical and electrophysiological methods. Horseradish peroxidase (HRP)-labelled clusters were observed in the periventricular nucleus after application of HRP into the VMH. Periventricular nucleus stimulation produced mainly excitation of VMH neurones with a latency of 3 msec. These results suggest that periventricular nucleus sends information in the cerebro-spinal fluid to the VMH. © 1982.

    DOI PubMed

  • Ontogenesis of glucose sensitivity in the rat lateral hypothalamus: a brain slice study

    Shigenobu Shibata, Yutaka Oomura, Hitoshi Kita

    Developmental Brain Research   5 ( 1 ) 114 - 117  1982  [Refereed]

     View Summary

    The development of glucose-sensitive neurons in the lateral hypothalamus (LHA) brain slices was examined. In 60-100-day-old rats, 20% of LHA neurons were sensitive, that is, their firing rates decreased upon increase in the glucose concentration in the perfusate and increased upon decrease in glucose concentration. In 2-, 3- and 7-day-old rats, 16% of the neurons were glucose sensitive. Results confirm glucose-sensitivity in the LHA at a very early stage of ontogeny. © 1982.

    DOI

  • Effects of medial amygdaloid lesions on the initiation and the maintenance of muricide in olfactory bulbectomized rats

    Shigenobu Shibata, Danny Suwandi, Tsuneyuki Yamamoto, Showa Ueki

    Physiology and Behavior   29 ( 5 ) 939 - 941  1982  [Refereed]

     View Summary

    In order to elucidate the role of medial amygdala on the initiation and the maintenance of muricide following olfactory bulb lesions, bilateral medial amygdaloid lesions were performed after, prior to, or simultaneously with olfactory bulb lesions. Both the initiation and the maintenance of muricide following olfactory bulb lesions were suppressed by amygdaloid lesions, and this result suggests that the medial amygdala plays a facilitatory role in both the initiation and the maintenance of muricide in olfactory bulbectomized rats. © 1982.

    DOI PubMed

  • Changes in amygdaloid afterdischarges and kindling effect following olfactory bulbectomy in the rat

    Shigenori Watanabe, Hiroshi Nakanishi, Shigenobu Shibata, Showa Ueki

    Physiology and Behavior   28 ( 4 ) 687 - 692  1982  [Refereed]

     View Summary

    Changes in the thresholds for afterdischarges and the formation of kindling effect in the medial amygdala following olfactory bulbectomy were investigated in the rat with chronic electrode implants. The threshold for afterdischarges in the amygdala of the olfactory bulbectomized rat (OB rat) was significantly decreased on day 4 after olfactory bulbectomy, however, no significant difference was found between OB and sham operated rats on days 7, 14 and 21 since the threshold in the sham group was also decreased at these periods after the surgery. The formation of kindling effect was remarkably accelerated in the OB rats. In this case, the number of days required to reach the stage 1 (Racine's classification) was significantly shortened. These results suggest that the activity of the medial amygdaloid nucleus is increased following olfactory bulbectomy. © 1982.

    DOI PubMed

  • Differential effects of medial, central and basolateral amygdaloid lesions on four models of experimentally-induced aggression in rats

    Shigenobu Shibata, Tsuneyuki Yamamoto, Showa Ueki

    Physiology and Behavior   28 ( 2 ) 289 - 294  1982  [Refereed]

     View Summary

    To clarify whether various nuclei of the amygdaloid complex play different roles in aggressive behavior including muricide, 4 types of aggression were experimentally induced in rats. These include olfactory bulbectomy (OB rats), midbrain raphe lesions (Raphe rats), administration of Δ9-tetrahydrocannabinol (THC rats) and long-term isolation (Iso rats). Rats which exhibited muricide following these treatments were subjected to bilateral lesions of either the medial (AME), central (ACE) or basolateral (ABL) amygdaloid nuclei. Both muricide and hyperemotionality in the OB rat were markedly inhibited by AME lesions. Those of the Iso and THC rats were moderately inhibited. However, in the Raphe rat, aggressive behavior was not inhibited by AME lesions. Furthermore, ACE or ABL lesions caused no significant changes in all 4 models of aggression. These results suggest that the AME plays a facilitatory role in aggression of OB, Iso and THC rats, but aggression in Raphe rat is independent of amygdaloid activity. © 1982.

    DOI PubMed

  • Behavioral effects of flutoprazepam (KB-509) and its metabolites

    Showa Ueki, Takayuki Sukamoto, Shigenori Watanabe, Tsuneyuki Yamamoto, Yasufumi Kataoka, Shigenobu Shibata, Danny Suwandi, Kazuhiko Shibata, Masako Takano, Yoko Sato

    Folia Pharmacologica Japonica   80 ( 1 ) 15 - 30  1982  [Refereed]

     View Summary

    The behavioral effects of KB-509 and its metabolites were investigated and compared with those of diazepam in mice and rats. The locomotor activity of mice measured by an Animex test was decreased with relatively large doses of KB-509 and diazepam. The anticonflict effect of KB-509 in rats was approximately as potent as that of diazepam. The lever pressing responses in the unpunished period were reduced by diazepam at a dose of 50 mg/kg p.o., while they were not affected by KB-509 even at a large dose such as 100 mg/kg p.o. KB-509 inhibited conditioned avoidance responses of the rat in a shuttle box at an extremely large dose such as 1000 mg/kg p.o. This effect of KB-509 was much less potent than that of diazepam. KB-509 was more potent than diazepam in inhibiting footshock-induced fighting behavior in mice. KB-509 was more potent than diazepam in suppressing hyperemotionality, but was less potent than diazepam in inhibiting muricide of olfactory bulbectomized rats. KB-509 and diazepam prevented maximal electroshock, pentetrazol, and strychnine induced convulsions in mice. KB-509 was more potent than diazepam in potentiating barbital anesthesia, in impairing rotarod performance, and in muscle relaxant activity measured by a traction test in mice. The pharmacological activity of desalkyl-KB-509 was more potent than that of KB-509 and that of desalkyl-3-OH-KB-509 was approximately as potent as that of KB-509. These results indicate that KB-509 possesses pharmacological properties similar to that of diazepam, and it is slightly greater in potency and is much longer in duration of action than diazepam. © 1982, The Japanese Pharmacological Society. All rights reserved.

    DOI PubMed

  • Effects of Locus Coeruleus Stimulation on Muricide in Olfactory Bulbectomized and Raphe Lesioned Rats

    Tsuneyuki Yamamoto, Shigenobu Shibata, Showa Ueki

    The Japanese Journal of Pharmacology   32 ( 5 ) 845 - 853  1982  [Refereed]

     View Summary

    In order to elucidate the role of central monoamines in muricide of the rat, the effects of electrical stimulation of the locus coeruleus (LC) were investigated on two types of muricide induced by olfactory bulbectomy (OB rats) and midbrain raphe lesions (raphe rats). Muricide was inhibited in 71.4% of the OB rats by bilateral LC stimulation and in 26.7% by unilateral stimulation. Even in the rat in which muricide was not inhibited following LC stimulation, muricide was almost invariably suppressed by LC stimulation after pretreatment with pargyline. The antimuricidal effect of LC stimulation was partially blocked by administration of propranolol, but not by phenoxy benzamine. In contrast, muricide was inhibited by bilateral LC stimulation in 44.4% of the raphe rats, but this effect was not potentiated by pretreatment with pargyline. On the other hand, muricide was not significantly inhibited by either dorsal raphe or medial raphe stimulation in any OB rats. These results suggest that noradrenaline plays a more important role in inhibiting muricide in OB rats than in raphe rats. © 1982, The Japanese Pharmacological Society. All rights reserved.

    DOI PubMed

  • Effects of electroconvulsive shock on mouse-killing behavior (muricide) in olfactory bulbectomized rats

    Shigenobu Shibata, Shigenori Watanabe, Hiroshi Nakanishi, Showa Ueki

    The Japanese Journal of Pharmacology   31 ( 2 ) 275 - 280  1981  [Refereed]

     View Summary

    We investigated the influence of electroconvulsive shock (ECS), regarded to possess an antidepressant effect clinically, on muricide in olfactory bulbectomized rats (0B rats). Muricide in these rats was markedly inhibited by ECS treatment. Five and 10 min after the termination of ECS-induced convulsions, muricide was inhibited by 100%. Even after intervals of 20 and 60 min, inhibition rates of 80% and 30% were obtained, respectively. ECS-induced muricide inhibition was remarkably antagonized by pretreatment with the α-blocker phenoxybenzamine but not by pretreatment with the β-blocker sotalol. ECS-induced suppression of muricide was potentiated by repeated ECS treatment once daily for 10 days. After several applications of ECS treatment, muricide was inhibited in muricide tests done 24 hours after ECS treatment
    this state persisted for up to 10 days thereafter. The results of this experiment demonstrated that ECS treatment specifically inhibited muricide in OB rats and further suggested that the cerebral noradrenergic α-receptor system plays an important role in this ECS-induced inhibition of muricide. Similar to findings in the case of antidepressant administration, inhibition of muricide was potentiated by chronic ECS treatment. Specific inhibition of muricide in OB rats by antidepressants indicated that this phenomenon may serve as an animal model for the evaluation of antidepressant activity. Our results reiterate the usefulness of muricide in OB rats as an excellent experimental model for the assessment of antidepressant activity. © 1981, The Japanese Pharmacological Society. All rights reserved.

    DOI PubMed

  • Behavioral and electroen cephalographic effects of alprazolam and its metabolites

    Showa Ueki, Shigenori Watanabe, Tsuneyuki Yamamoto, Yasufumi Kataoka, Naomori Tazoe, Shigenobu Shibata, Kazuhiko Shibata, Hisashi Ohta, Kiminori Kawahara, Masako Takano, Danny Suwandi, Soon Chul Lee, Shy Yuh Liou

    Folia Pharmacologica Japonica   77 ( 5 ) 483 - 509  1981  [Refereed]

     View Summary

    The behavioral and electroencephalographic effects of alprazolam and its metabolites were investigated in mice, rats and rabbits, and compared with the data on diazepam, lorazepam and nitrazepam. Locomotor activity of mice in an open-field situation was increased with smaller doses of alprazolam and nitrazepam but not with diazepam and lorazepam. Alprazolam increased the hyperactivity induced by methamphetamine in mice. The anticonflict effects of alprazolam in rats were more potent than those of diazepam and lorazepam. In suppressing hyperemotionality and muricide of olfactory bulbectomized rats, alprazolam was more potent than diazepam and fairly equal to that of lorazepam. Muricide of raphe-lesioned rats was markedly inhibited by alprazolam. Mescaline-induced head-twitches in mice were more markedly increased with alprazolam than with nitrazepam and diazepam. Alprazolam, diazepam and lorazepam prevented both maximal electroshock and pentetrazol convulsions in mice. Alprazolam was more potent than diazepam and lorazepam in potentiating thiopental, ether and ethanol anesthesia in mice. In the impaired rotarod performance, alprazolam was more potent than diazepam and lorazepam in mice, but was much less potent than lorazepam in rats. In conscious rabbits with chronically implanted electrodes, alprazolam induced a drowsy EEG pattern and depressed the EEG arousal response not only to auditory stimulation but also to mesencephalic reticular stimulation. The EEG effect of alprazolam was approx. twice that of diazepam. The pharmacological activity of a-hydroxyalprazolam was approx. one third that of alprazolam. However, 5-chloro-2-(3-hydromethy1-5-methyl-4H-1, 2,4-triazolo-4-y1) benzophenone showed no pharmacological activity. These results indicate that alprazolam possesses pharmacologic properties characteristic to benzodiazepines and that the activity is more potent than diazepam. In addition, alprazolam seems to have a specific effect on the central seroto nergic mechanisms. © 1981, The Japanese Pharmacological Society. All rights reserved.

    DOI PubMed

  • Development of Experimental Setup to Create Novel Mental Disorder Model Rats Using Small Mobile Robot

    Hiroyuki Ishii, Qing Shi, Atsuo Takanish, Satoshi Okabayashi, Naritoshi Iida, Hiroshi Kimura, Yu Tahara, Akiko Hirao, Shigenobu Shibata

    The 2010 IEEE/RSJ International Conference on Intelligent Robots and Systems (IROS2010)  

  • A novel method to develop an animal model of depression using a small mobile robot

    Hiroyuki Ishii, Qing Shi, Shogo Fumino, Shinichiro Konno, Satoshi Okabayashi, Naritoshi Iida, Hiroshi Kimura, Yu Tahara, Shigenobu Shibata, Atsuo Takanish

    Advanced Robotics  

▼display all

Books and Other Publications

  • イラストで徹底理解するシグナル伝達キーワード事典「サーカディアンシグナリング」

    堀川和政, 柴田重信

    羊土社  2012.08 ISBN: 9784758120333

  • 時間生物学「概日リズムと代謝」

    柴田重信

    化学同人  2012.03 ISBN: 9784759815023

  • 体内時計の科学と産業応用 第11章「食品の体内時計に対する効果」

    平尾彰子, 柴田重信

    CMC出版  2011.05 ISBN: 9784781303406

  • 体内時計の科学と産業応用 第12章「体内時計が栄養・食物摂取に及ぼす効果」

    柴田重信

    CMC出版  2011.05 ISBN: 9784781303406

  • 実践行動薬理学 pp27-35「概日リズム評価法」

    田原 優, 柴田重信

    金芳堂  2010.03 ISBN: 9784765314107

  • Memory and Sleep[Encyclopedia of Neuroscience]

    Shibata S

    Springer publication  2009.02

  • 時間栄養学

    香川靖雄, 柴田重信, 小田裕昭, 加藤秀夫, 堀江修一, 榛葉繁紀

    女子栄養大学出版部  2009.02 ISBN: 9784789554336

  • 時間生物学事典

    柴田重信

    朝倉書店  2008.05 ISBN: 9784254171303

  • 生物物理学ハンドブック

    柴田重信

    朝倉書店  2007.05 ISBN: 9784254171228

  • 生物学的反応 時間薬理学の体内時計「リウマチ病セミナー15」

    柴田重信, 前田晃, 小松原良雄, 福田眞輔, 吉川秀樹, 志水正敏, 塩沢俊一, 村田紀和, 佐伯行彦, 西林保朗

    永井書店  2005.01 ISBN: 9784815917074

  • 時計遺伝子の分子生物学 P.139-146

    柴田重信, 岡村均, 深田吉孝

    シュプリンガーフェアラーク東京  2004.04 ISBN: 9784431710233

  • 女と男の人間科学

    柴田重信

    コロナ社  2003.12 ISBN: 9784339078381

  • ニューロシグナリングから知識工学への展開 第8章「脳のリズム」

    柴田重信

    コロナ社  2002 ISBN: 9784339078350

  • 「時間薬理学」創薬の視点に立った生体リズム 14-22

    柴田重信, 秋山正志

    朝倉書店  2001 ISBN: 9784254340112

  • Per gene expression and circdian entrainment by photic,non-photic and daily scheduled feeding, In, Zeitogebers, entrainment and masking of the circadian system

    Shibata,S, Horikawa,K, Yokota,S, Fuji,K, Akiyama,M, Yoshinobu,Y, Wakamatsu,H, Moriya, T, jointly worked

    Hokkaido Univ. Press  2001

  • 「生物時計の分子生物学」細胞内情報伝達

    柴田重信, 守屋孝洋

    シュプリンガー・フェアラーク東京  1999 ISBN: 9784431708148

  • 「生体リズムと精神疾患 (生物学的精神医学)」時計機構の老化

    柴田重信, 浜田俊幸

    学会出版センター  1997 ISBN: 9784762208430

  • Neurotransmitter interaction in the suprachiasmatic nucleus clock function. Evolution of circadian clock

    Shibata,S, Hamada,T, Watanabe,A, Ono,M, Watanabe,S, jointly worked

    Hokkaido Univ. Press  1994 ISBN: 4832902059

  • Role of 5-HT1A receptor mechanism in the circadian systems: assessed by wheel-running behavior, 2-deoxyglucose uptake and firing discharge in the rodent suprachiasmatic nuclei. In [New functional aspects of the suprachiasmatic nucleus of the hypothalamus]

    Shibata,S, Tominaga,K, Hamada,T, Tsuneyoshi,A, Watanabe,S, jointly worked

    JohonLibbey Press, London  1993 ISBN: 9780861963294

  • Circadian rhythm of suprachiasmatic nucleus neurons and its effects on feeding. In [New functional aspects of the suprachiasmatic nucleus of the hypothalamus]

    Oomura,Y, Shibata,S, jointly worked

    JohonLibbey Press, London  1993 ISBN: 9780861963294

  • Developmental anatomy of the circadian system. In [Development of circadian rhythmicity and photoperiodism in mammals]

    Moore,RY, Shibata,S, Bernstein ME, jointly worked

    Perinatology Press  1990 ISBN: 9780916859428

  • 「医薬品の開発 第8巻」薬物の作用機構 抗うつ薬

    植木昭和, 柴田重信

    廣川書店  1989 ISBN: 9784567390804

  • Neurophysiological and neurochemical studies of rodent suprachiasmatic nucleus in vitro. Circadian clocks and ecology

    Shibata, S, Moore,RY, jointly worked

    Hokkaido Univ. press  1989 ISBN: 4832902032

  • 特集・サーカディアンリズム-基礎から臨床へ-「精神医学(31巻1)」

    高橋三郎, 井上慎一, 高橋清久, 本間研一, 柴田重信ほ

    医学書院  1989

  • うつ病と脳内アミンーラットのmuricideについての研究「続 脳の生体警告 不安・不快機構」

    植木昭和, 藤原道弘, 山本経之, 片岡泰文, 柴田重信, 岩崎克典

    東京大学出版  1987 ISBN: 9784130601177

  • Electrphysiological bases for rhythmic activity in the suprachiasmatic nucleus of the rat: an in vitro study. Emotions, neuronal and humoral control

    Sugimori,M, Shibata,S, Oomura Y, jointly worked

    Japan Scientific Society press- S Karger press  1986

  • Observations of suprachiasmatic neuronal activity in relation to circadian clocks and zeitgebers

    Shibata,S, Oomura, Y."jointly worked

    Hokkaido Univ. Press  1985

  • Circadian rhythm of suprachiasmatic nucleus neurons and its control to feeding centers in the rat hypothalamus.

    Oomura,Y, Shibata,S, Kita,H."jointly worked

    Pergamon Press, Oxford and New York  1982 ISBN: 9780080279770

▼display all

Misc

  • Effects of time-restricted feeding on the mitochondrial DNA mutator mice (POLG)

    李楠, 大池秀明, 田原優, 柴田重信

    時間生物学   26 ( 2 )  2020

    J-GLOBAL

  • ウロリチンAのPer2遺伝子発現リズムに対するインビトロ評価

    杜堯, 折原芳波, 原口敦嗣, 立石法史, 柴田重信

    日本栄養・食糧学会大会講演要旨集   74th  2020

    J-GLOBAL

  • The effects of Polygalae Radix on clock gene expression rhythm and free-running period

    原口敦嗣, 斉藤恵祐, 田原優, 柴田重信

    時間生物学   26 ( 2 )  2020

    J-GLOBAL

  • Evaluate the effects of light, feeding, and stress that cause disturbance in peripheral clocks and locomotor activity rhythms due to social jetlag

    原口敦嗣, 田村好, 佐藤修平, 山崎智弘, 西村裕太郎, 福澤雅, 柴田重信

    時間生物学   25 ( 2 )  2019

    J-GLOBAL

  • Recovery effects of combination of caffeine and histidine on circadian clock system disturbed by social jetlag

    尾根田諭, 原口敦嗣, 石井裕也, 山崎智弘, 田村好, 佐藤脩平, 柴田重信

    時間生物学   25 ( 2 )  2019

    J-GLOBAL

  • 体内時計研究―基礎から社会応用に向けて―1 体内時計を考慮した時間栄養学と時間運動学による健康づくり 栄養と運動による健康増進を時間軸で捉える

    青山晋也, 青山晋也, 柴田重信

    化学と生物   57 ( 1 ) 43‐49  2018.12  [Invited]

    Article, review, commentary, editorial, etc. (scientific journal)  

    J-GLOBAL

  • Relationship Between Clock Gene Expression, MEQ Score, and Exercise Performance

    Karina Ando, Masaki Takahashi, Shigenobu Shibata, Hideyuki Takahashi

    MEDICINE AND SCIENCE IN SPORTS AND EXERCISE   50 ( 5 ) 793 - 793  2018.05

    Research paper, summary (international conference)  

  • マウスを用いたカフェインの抗肥満効果に関する時間栄養学的研究

    山崎 智弘, 原口 敦嗣, 西村 祐太郎, 大久保 仁, 佐々木 裕之, 田村 好, 志賀 一登, 柴田 重信

    日本栄養・食糧学会大会講演要旨集   72回   280 - 280  2018.04

  • 医療のさらなる安心・安全を志向する若手研究者のイノベーティブチャレンジ 時計遺伝子Per2の発現周期やマウス行動周期に対する遠志の影響について

    原口 敦嗣, 中村 文彬, 阿部 真太郎, 中尾 洋一, 柴田 重信

    日本薬学会年会要旨集   138年会 ( 1 ) 336 - 336  2018.03

  • Chrono-Nutritional Study of the Eff ect of Soy Protein Isolate and Soy Isofl avone on Muscle Hypertrophy in Mice

    Shinya Aoyama, Rina Hirooka, Shigenobu Shibata

      39 ( 21 ) 167 - 172  2018

    Meeting report  

  • 社会と共創するスポーツ科学研究の展開 2―時間栄養学からみた運動のあり方

    青山晋也, 柴田重信

    体育の科学   68 ( 1 ) 4‐8  2018.01  [Invited]

    Article, review, commentary, editorial, etc. (scientific journal)  

    J-GLOBAL

  • 「時間栄養学」でメタボを予防 最終回 IoT・スマホ時代の時間栄養学

    青山晋也, 柴田重信, 柴田重信

    安全と健康   68 ( 6 ) 602‐603 - 603  2017.06  [Invited]

    Article, review, commentary, editorial, etc. (trade magazine, newspaper, online media)  

    J-GLOBAL

  • わが国の機能性食品の科学と産業の興隆を目指して〈2〉(前編)時間栄養学・時間運動学と健康増進

    青山晋也, 柴田重信

    生物工学会誌   95 ( 5 ) 248‐251  2017.05  [Invited]

    Article, review, commentary, editorial, etc. (scientific journal)  

    J-GLOBAL

  • フラボノイド系化合物の培養細胞下における体内時計への効果

    篠崎 綾子, 三澤 憲一郎, 池田 祐子, 原口 敦嗣, 鎌形 真世, 田原 優, 柴田 重信

    日本栄養・食糧学会大会講演要旨集   71回   339 - 339  2017.04

  • Potent effects of the flavonoid nobiletin on the amplitude, period, and phase of the circadian clock rhythm in PER2::LUCIFERASE mouse embryonic fibroblasts

    Ayako Shinozaki, Kenichiro Misawa, Yuko Ikeda, Atsushi Haraguchi, Mayo Kamagata, Yu Tahara, Shigenobu Shibata

    JOURNAL OF PHARMACOLOGICAL SCIENCES   133 ( 3 ) S204 - S204  2017.03

    Research paper, summary (international conference)  

  • 「時間栄養学」でメタボを予防 メタボ予防の時間栄養・運動学

    青山 晋也, 柴田 重信

    安全と健康   68 ( 3 ) 290 - 291  2017.03  [Invited]

    Article, review, commentary, editorial, etc. (trade magazine, newspaper, online media)  

  • The Role of Circadian Rhythms in Muscular and Osseous Physiology and Their Regulation by Nutrition and Exercise

    Shinya Aoyama, Shigenobu Shibata

    FRONTIERS IN NEUROSCIENCE   11  2017.02

    Book review, literature introduction, etc.  

     View Summary

    The mammalian circadian clock regulates the day and night cycles of various physiological functions. The circadian clock system consists of a central clock in the suprachiasmatic nucleus (SCN) of the hypothalamus and peripheral clocks in peripheral tissues. According to the results of circadian transcriptomic studies in several tissues, the majority of rhythmic genes are expressed in a tissue-specific manner and are influenced by tissue-specific circadian rhythms. Here we review the diurnal variations of musculoskeletal functions and discuss the impact of the circadian clock on homeostasis in skeletal muscle and bone. Peripheral clocks are controlled by not only photic stimulation from the central clock in the SCN but also by external cues, such as feeding and exercise. In this review, we discuss the effects of feeding and exercise on the circadian clock and diurnal variation of musculoskeletal functions. We also discuss the therapeutic potential of chrono-nutrition and chrono-exercise on circadian disturbances and the failure of homeostasis in skeletal muscle and bone.

    DOI

  • 「時間栄養学」でメタボを予防 時間運動学とは

    青山 晋也, 柴田 重信

    安全と健康   68 ( 2 ) 186 - 187  2017.02  [Invited]

    Article, review, commentary, editorial, etc. (trade magazine, newspaper, online media)  

  • EFFECTS OF TIME-RESTRICTED PROTEIN FEEDING ON SKELETAL MUSCLE HYPERTROPHY IN MICE

    Aoyama Shinya, Tanaka Mizuho, Shimoda Takeru, Kojima Shuichi, Sasaki Keisuke, Shibata Shigenobu

    ANNALS OF NUTRITION AND METABOLISM   71   478  2017  [Refereed]

    Research paper, summary (international conference)  

  • 時間栄養学

    青山晋也, 柴田重信

    月刊内分泌・糖尿病・代謝内科   43 ( 6 ) 514‐517 - 517  2016.12  [Invited]

    Article, review, commentary, editorial, etc. (scientific journal)  

    CiNii J-GLOBAL

  • EGCGの投与時刻の違いが糖負荷後の血糖値変動に及ぼす影響

    高橋 将記, 坪坂 美来, 原口 敦嗣, 池田 祐子, 柴田 重信

    日本栄養・食糧学会大会講演要旨集   70回   165 - 165  2016.04

  • 夜食症候群モデルマウスにおける脳内モノアミン概日リズム変動の特徴

    福澤 雅, 原口 敦嗣, 西村 裕太郎, 岩見 志保, 本橋 弘章, 安田 晋之介, 柴田 重信

    日本栄養・食糧学会大会講演要旨集   70回   177 - 177  2016.04

  • Flavonoids uniquely affect circadian rhythm in vitro MEF from PER 2:: LUCIFERASE mouse

    Ayako Shinozaki, Kenichiro Misawa, Yuko Ikeda, Atsushi Haraguchi, Mayo Kamagata, Yu Tahara, Shigenobu Shibata

    JOURNAL OF PHARMACOLOGICAL SCIENCES   130 ( 3 ) S188 - S188  2016.03

    Research paper, summary (international conference)  

  • Inhibition of IgE-Mediated Allergic Reaction By Pharmacologically Targeting the Circadian Clock

    Yuki Nakamura, Atsuhito Nakao, Shigenobu Shibata

    JOURNAL OF ALLERGY AND CLINICAL IMMUNOLOGY   137 ( 2 ) AB76 - AB76  2016.02

    Research paper, summary (international conference)  

  • Social jetlagの現状と課題 社会的時間と生体リズムの不調和 食事時間帯とsocial jetlag

    原口 敦嗣, 西村 裕太郎, 柴田 重信

    日本睡眠学会定期学術集会プログラム・抄録集   40回   110 - 110  2015.07

  • 人工赤血球製剤の効力と安全性 妊娠高血圧症候群に対する人工赤血球を用いた治療法の開発

    太田 英伸, 李 コウ, 中川 真智子, 若松 永憲, 泉 仁美, 稲垣 真澄, 岡村 州博, 小田切 優樹, 横田 秀夫, 柴田 重信, 酒井 宏水, 八重樫 伸生

    人工臓器   43 ( 2 ) S - 57  2014.09

  • 食行動と概日リズム調節機構 (特集 睡眠・眠りの基礎と臨床)

    柴田 重信, 池田 祐子, 原口 敦嗣

    医薬ジャーナル   50 ( 6 ) 69 - 73  2014.06

    CiNii

  • 【睡眠・眠りの基礎と臨床】 食行動と概日リズム調節機構

    柴田 重信, 池田 祐子, 原口 敦嗣

    医薬ジャーナル   50 ( 6 ) 1555 - 1559  2014.06

     View Summary

    視交叉上核(SCN)は、哺乳類における体内の生理現象を約24時間周期に調節している。食行動ホルモンと体内時計の研究は、マウスやラットを用いて盛んに行われている。オレキシンやグレリン等の食欲増進ホルモンや、レプチン等の食欲抑制ホルモンも、SCNの支配を受けて日内リズムを刻むことで、食行動にもリズム性が生じることが分かってきた。さらにヒトを対象とした研究により、食事の摂り方によって食欲ホルモン分泌が変化し、肥満に関与していることが明らかとなった。近年、ヒトの体内時計の測定手法が確立されたことから、ヒトを対象とした食欲ホルモンと体内時計の研究の発展が期待される。(著者抄録)

  • Hippocampal neurogenesis disorder and mood disorder-like behavior in clock mutant mice

    Jun Takoda, Hiromu Fukuzawa, Tokiko Suzuki, Shigenobu Shibata, Takahiro Moriya

    JOURNAL OF PHARMACOLOGICAL SCIENCES   124   128P - 128P  2014

    Research paper, summary (international conference)  

  • Restraint stress strongly disturbs the mouse circadian clock systems

    Yu Tahara, Takuya Shiraishi, Yosuke Kikuchi, Nobuaki Onishi, Shun-ya Yamada, Atsushi Haraguchi, Daisuke Kuriki, Shigenobu Shibata

    JOURNAL OF PHARMACOLOGICAL SCIENCES   124   206P - 206P  2014

    Research paper, summary (international conference)  

  • Circadian rhythm of inflammatory reaction and chrono-pharmacological treatment to the ulcerative colitis in mice

    Yosuke Kikuchi, Yu Tahara, Takuya Shiraishi, Shigenobu Shibata

    JOURNAL OF PHARMACOLOGICAL SCIENCES   124   206P - 206P  2014

    Research paper, summary (international conference)  

  • Study of evaluation method of natural killer cell activity in mice with flow cytometry

    Yosuke Kikuchi, Yu Tahara, Takuya Shiraishi, Shigenobu Shibata

    JOURNAL OF PHARMACOLOGICAL SCIENCES   124   206P - 206P  2014

    Research paper, summary (international conference)  

  • 拘束ストレスは強い末梢時計同調効果を持つ

    白石卓也, 田原優, 菊池耀介, 大西信明, 原口敦嗣, 栗城大輔, 柴田重信

    時間生物学   19 ( 2 ) 164  2013.10

    J-GLOBAL

  • chrono-nutrition and psychological health

    Food style 21   17 ( 8 ) 42 - 44  2013.08

    CiNii

  • 高脂肪食の摂取時刻や摂取タイミングによる体重や脂肪率、末梢時計への影響

    原口 敦嗣, 青木 菜摘, 大津 定治, 柴田 重信

    日本栄養・食糧学会大会講演要旨集   67回   213 - 213  2013.04

  • マウスを用いた時間栄養学視点からの塩分摂取と排泄機構の研究

    柴田重信, 青木菜摘, 濱口雄太郎, 原口敦嗣

    ソルト・サイエンス研究財団助成研究報告集 2 医学 食品科学編   2011   229 - 236  2013.03

    J-GLOBAL

  • Combination food of carbohydrate and protein entrains mouse peripheral clocks through insulin secretion

    Yuko Ikeda, Teiji Ohtsu, Daisuke Kuriki, Akiko Hirao, Shigenobu Shibata

    JOURNAL OF PHYSIOLOGICAL SCIENCES   63   S282 - S282  2013

    Research paper, summary (international conference)  

  • High fat diet feeding duration and time dependent impairment of energy homeostasis and peripheral clock phase

    Atsushi Haraguchi, Natsumi Aoki, Teiji Ohtsu, Shigenobu Shibata

    JOURNAL OF PHYSIOLOGICAL SCIENCES   63   S282 - S282  2013

    Research paper, summary (international conference)  

  • The influence of soyaflavone HG on the period of mouse clock gene expression in vitro and in vivo

    Mari Kuwahara, Keisuke Saito, Seira Narishige, Shigenobu Shibata

    JOURNAL OF PHARMACOLOGICAL SCIENCES   121   201P - 201P  2013

    Research paper, summary (international conference)  

  • シフトワークモデルマウスの様態観察と時間栄養学的治療法の確立

    田原優, 濱口雄太郎, 一杉真史, 黒田大暁, 原口敦嗣, 柴田重信

    日本分子生物学会年会プログラム・要旨集(Web)   35th   3W11III-3 (WEB ONLY)  2012

    J-GLOBAL

  • Stereoselective disposition of S- and R-[6]-gingerol in mice after oral administration

    Mayumi Okamoto, Keigo Kamikihara, Hiroyuki Irii, Yu Tahara, Akiko Hirao, Shigenobu Shibata, Isao Shimizu

    JOURNAL OF PHARMACOLOGICAL SCIENCES   115   74P - 74P  2011

    Research paper, summary (international conference)  

  • 血糖値上昇抑制/低下作用を有する[6]‐gingerol及びその類縁体の合成と生理活性評価

    岡本真由美, 入井啓行, 茂原隆久, 田原優, 平尾彰子, 柴田重信, 清水功雄

    日本糖尿病・肥満動物学会年次学術集会プログラム・講演抄録集   25th   56  2011

    J-GLOBAL

  • The effect of exercise on the phase of circadian rhythm and metabolic-related genes in mice

    Karina Ando, Akiko Hirao, Sunao Uchida, Shigenobu Shibata

    JOURNAL OF PHYSIOLOGICAL SCIENCES   60   S192 - S192  2010

    Research paper, summary (international conference)  

  • Research of mouse circadian rhythm based on chrononutrition approach

    Shigenobu Shibata

    JOURNAL OF PHYSIOLOGICAL SCIENCES   60   S14 - S14  2010

    Research paper, summary (international conference)  

  • Effect of Chinese herb Jyuzen-taiho-to on clock gene and metabolism related genes in the mouse liver

    Goki Koiwa, Yu Tahara, Akiko Hirao, Takeshi Morishima, Shigenobu Shibata

    JOURNAL OF PHARMACOLOGICAL SCIENCES   112   167P - 167P  2010

    Research paper, summary (international conference)  

  • A relationship between circadian rhythm and pathogenesis of atopic dermatitis-like skin lesions in mice

    Eriko Takita, Yuji Kubo, Yu Tahara, Atsuhito Nakao, Shigenobu Shibata

    JOURNAL OF PHYSIOLOGICAL SCIENCES   60   S198 - S198  2010

    Research paper, summary (international conference)  

  • Chronopharmacological and molecular studies of anti-diuretic drugs on Per2 mutant mice

    Naoki Furutani, Yu Tahara, Akiko Hirao, Shigenobu Shibata

    JOURNAL OF PHARMACOLOGICAL SCIENCES   112   154P - 154P  2010

    Research paper, summary (international conference)  

  • BALANCED DIETS WITH HIGH GLYCEMIC INDEX, BUT NOT THOSE WITH SUGAR ALONE, ARE SUITABLE ENTRAINMENT SIGNALS FOR MOUSE LIVER CLOCK

    Shigenobu Shibata, Akiko Hirao, Yu Tahara

    ANNALS OF NUTRITION AND METABOLISM   55   457 - 457  2009

    Research paper, summary (international conference)  

  • The dorsomedial hypothalamus is not necessary for food-associated rhythms of behavior, temperature or clock gene expression in mice

    Takahiro Moriya, Reiko Aida, Takashi Kudo, Masashi Akiyama, Masao Doi, Naomi Hayasaka, Ralph Mistlberger, Hitoshi Okamura, Shigenobu Shibata, Norimichi Nakahata

    NEUROSCIENCE RESEARCH   65   S231 - S231  2009

    Research paper, summary (international conference)  

    DOI

  • Behavioral alterations in prednisolone-treated mice

    Yu Kajiyama, Miyako Furuta, Midori Ninomiya, Shigenobu Shibata, Hiroshi Kunugi

    NEUROSCIENCE RESEARCH   61   S135 - S135  2008

    Research paper, summary (international conference)  

  • Chronic social defeat stress induces depression-like behavior and apoptosis in hippocampus

    Midori Ninomiya, Miyako Furuta, Yu Kajiyama, Shigenobu Shibata, Hiroshi Kunugi

    NEUROSCIENCE RESEARCH   61   S206 - S206  2008

    Research paper, summary (international conference)  

  • Autonomic nerve regulates circadian clock gene expression in the mouse lung with nicotine pretreatment

    Shigenobu Shibata, Yuta Motoyama, Shun Shiota, Takashi Kudo, Kazumasa Horikawa

    JOURNAL OF PHARMACOLOGICAL SCIENCES   103   257P - 257P  2007

    Research paper, summary (international conference)  

  • PPAR alpha is a potential therapeutic drug target of circadian rhythm sleep disorders

    Katsutaka Oishi, Hidenori Shirai, Takashi Kudo, Shigenobu Shibata, Norio Ishida

    NEUROSCIENCE RESEARCH   58   S48 - S48  2007

    Research paper, summary (international conference)  

  • Effect of mood disorder therapeutic drugs on mice circadian clock systems

    Takashi Kudo, Akiko Yoshida, Syoutaro Yamaguchi, Kazumasa Horikawa, Shigenobu Shibata

    JOURNAL OF PHARMACOLOGICAL SCIENCES   103   136P - 136P  2007

    Research paper, summary (international conference)  

  • Neuronal and hormonal control of peripheral clock function through suprachiasmatic nucleus

    Shigenobu Shibata, Naomi Hayasaka, Takashi Kudo, Tsuyoshi Yaita

    NEUROSCIENCE RESEARCH   55   S44 - S44  2006

    Research paper, summary (international conference)  

  • Effects of dexamethasone on emotional behavior and gene expression in mice

    Tomoko Soga, Yu Kajiyama, Shigenobu Shibata, Hiroshi Kunugi

    NEUROSCIENCE RESEARCH   55   S93 - S93  2006

    Research paper, summary (international conference)  

  • Circadian oscillation of Ca2+/calmodulin-dependent protein kinase II activity in the hamster suprachiasmatic nucleus

    S Matsuda, T Hamada, S Shibata, K Fukunaga

    JOURNAL OF PHARMACOLOGICAL SCIENCES   97   91P - 91P  2005

    Research paper, summary (international conference)  

  • Identification of a circadian mechanism for gating photic input.

    T Hamada, R Silver, S Shibata

    JOURNAL OF PHARMACOLOGICAL SCIENCES   91   20P - 20P  2003

    Research paper, summary (international conference)  

  • 短時間作動型睡眠導入薬ブロチゾラムのハムスターの輪回し行動リズムとSCN内での Period mRNA の発現に対する効果

    横田伸一, 堀川和政, 秋山正志, 守屋孝洋, 海老原史樹文, 太田龍朗, 柴田重信

    日本時間生物学会会誌: Journal of Chronobiology   6 ( 2 )  2000.10

    CiNii

  • ラット視交叉上核におけるLTPの情報伝達系とメラトニンによる調節

    福永浩司, 笠原二郎, 柴田重信, 宮本英七

    日本時間生物学会会誌: Journal of Chronobiology   6 ( 2 )  2000.10

    CiNii

  • メタンフェタミンの薬理作用とmPer遺伝子発現

    二階堂隆人, 守屋孝洋, 柴田重信

    日本時間生物学会会誌: Journal of Chronobiology   5 ( 2 )  1999.10

    CiNii

  • ラット、マウスの老齢動物における Period mRNA 発現パターンの加齢による変化

    浅井良, 秋山正志, 南陽一, 二階堂隆人, 守屋孝洋, 柴田重信

    日本時間生物学会会誌: Journal of Chronobiology   5 ( 2 )  1999.10

    CiNii

  • 光刺激による概日リズムの位相変化に伴ったハムスター Period mRNA の発現分布

    堀川和政, 横田伸一, 守屋孝洋, 柴田重信

    日本時間生物学会会誌: Journal of Chronobiology   5 ( 2 )  1999.10

    CiNii

  • 光刺激によってSCN内で発現増強される PeriodmRNA に対するCaMキナーゼII

    横田伸一, 守屋孝洋, 柴田重信

    日本時間生物学会会誌: Journal of Chronobiology   5 ( 2 )  1999.10

    CiNii

  • 制限給餌におけるSCN非依存的給餌性リズムとmPer遺伝子の発現様式の解析

    若松永憲, 原礼子, 藍田礼子, 守屋孝洋, 柴田重信

    日本時間生物学会会誌: Journal of Chronobiology   5 ( 2 )  1999.10

    CiNii

  • マウスにおける周期的制限給餌による肝臓 Period mRNA 発現への影響について

    原礼子, 若松永憲, 守屋孝洋, 秋山正志, 柴田重信

    日本時間生物学会会誌: Journal of Chronobiology   5 ( 2 )  1999.10

    CiNii

  • ストレスによるmPer遺伝子発現調節

    高橋里美, 若松永憲, 守屋孝洋, 柴田重信

    日本時間生物学会会誌: Journal of Chronobiology   5 ( 2 )  1999.10

    CiNii

  • マウスSCNにおけるmCry1 mRNAの発現と行動リズムについて

    須藤元喜, 守屋孝洋, 柴田重信

    日本時間生物学会会誌: Journal of Chronobiology   5 ( 2 )  1999.10

    CiNii

  • 生物時計の分子生物学

    柴田重信, 守屋孝洋

    生物時計の分子生物学     178 - 188  1999

    Article, review, commentary, editorial, etc. (other)  

  • mPer遺伝子アンチセンスによるマウス概日性活動リズムの位相変化の抑制

    秋山正志, 高橋里美, 神津靖子, 若松永憲, 守屋孝洋, 柴田重信

    日本神経精神薬理学雑誌 = Japanese journal of psychopharmacology   18 ( 6 )  1998.12

    CiNii

  • メラトニン、メチルコバラミンの飲水投与による明暗サイクルの連続アドウァンスに対する再同調促進作用 : 行動リズムならびにmPer1 mRNA発現による評価

    浅井 良, 秋山 正志, 柴田 重信

    日本時間生物学会会誌: Journal of Chronobiology   4 ( 2 ) 107 - 107  1998.10

    CiNii

  • 光刺激で誘導される哺乳類per遺伝子ホモログの相互作用解析

    若松 永憲, 高橋 里美, 秋山 正志, 柴田 重信

    日本時間生物学会会誌: Journal of Chronobiology   4 ( 2 ) 126 - 126  1998.10

    CiNii

  • グルタミン酸による視交叉上核神経活動の位相変化に対するmPer1アンチセンスオリゴヌクレオチドの効果

    守屋孝洋, 秋山正志, 前谷幸, 渡辺繁紀, 柴田重信

    日本時間生物学会会誌: Journal of Chronobiology   4 ( 2 )  1998.10

    CiNii

  • メタンフェタミン惹起性予知行動リズムとmPer遺伝子

    二階堂隆人, 守屋孝洋, 秋山正志, 柴田重信

    日本時間生物学会会誌: Journal of Chronobiology   4 ( 2 )  1998.10

    CiNii

  • Ryanodine receptor type 3 (RyR3) ノックアウトマウスの行動リズム解析

    神津靖子, 守屋孝洋, 西美幸, 竹島浩, 柴田重信

    日本時間生物学会会誌: Journal of Chronobiology   4 ( 2 )  1998.10

    CiNii

  • Triazolam により誘発されるハムスターの輪回し行動リズムの位相前進作用に対するMKC-242の効果

    横田伸一, 守屋孝洋, 柴田重信

    日本時間生物学会会誌: Journal of Chronobiology   4 ( 2 )  1998.10

    CiNii

  • カルシウムイメージング法を用いた視交差上核NMDA受容体の機能解析

    池田 真行, 浅井 良, 中村 行宏, 角谷 寛, 中西 重忠, 吉岡 亨, 柴田 重信

    日本時間生物学会会誌: Journal of Chronobiology   3 ( 2 )  1997.10

    Research paper, summary (national, other academic conference)  

    CiNii

  • アストログリア特異的に発現する Glial Fibrillary Acidic Protein (GFAP) の概日リズムにおける役割について

    吉信ゆう子, 川崎美帆, 神津靖子, 守屋孝洋, 池田真行, 吉岡亨, 加藤明, 五味浩司, 糸原重美, 柴田重信

    日本時間生物学会会誌: Journal of Chronobiology   3 ( 2 )  1997.10

    CiNii

  • NMDA受容体サブタイプノックアウト動物に対する制限給餌同調の効果

    神津靖子, 守屋孝洋, 池田真行, 高橋里美, 角谷寛, 中西重忠, 吉岡亨, 柴田重信

    日本時間生物学会会誌: Journal of Chronobiology   3 ( 2 )  1997

    CiNii

  • 体内時計におけるNMDA受容体サブタイプの役割

    高橋里美, 守屋孝洋, 池田真行, 秋山正志, 浅井良, 角谷寛, 中西重忠, 吉岡亨, 柴田重信

    日本時間生物学会会誌: Journal of Chronobiology   3 ( 2 )  1997

    CiNii

  • 虚血性神経細胞障害に対するvasopressinの作用

    越智まゆみ, 小泉修一, 柴田重信, 渡辺繁紀

    日本薬学会年会要旨集   112th ( Pt 3 )  1992

    J-GLOBAL

  • 制限給餌によるラットの時刻認知学習 老化,きゅう球摘出,中枢コリン低下の影響

    柴田重信, 恒吉厚子, 森永孝祐, 小泉修一, 渡辺繁紀, 皆本義基

    日本薬学会年会要旨集   112th ( Pt 3 )  1992

    J-GLOBAL

  • 新しい抗うつ薬Quinupramineの行動薬理学的研究(共著)

    植木昭和, 山本経之, 島添隆雄, 柴田重信, 谷吉弘, 町田幸一, 北条雅一, 吉田洋一, 辰巳ひろし

    日薬理誌   91 ( 6 ) 359 - 369  1988

    DOI PubMed

▼display all

Awards

  • 安藤百福学術大賞

    2016  

Research Projects

  • 時間栄養学を視点とした機能性食品成分の探索

    科学研究費助成事業(早稲田大学)  科学研究費助成事業(基盤研究(A))

  • 時間栄養学を視点とした機能性食品成分の探索と応用研究

    科学研究費助成事業(早稲田大学)  科学研究費助成事業(基盤研究(S))

  • 週間リズムの視点の運動科学・栄養科学による肥満予防

    科学研究費助成事業(早稲田大学)  科学研究費助成事業(挑戦的萌芽研究)

  • The roles of central neurotransmitters in the mechanism of action of psychotropic drugs.

  • Pharmacological studies on dementia

  • Development of effective drugs on impairments of Circardian rhythms and anticipatory behaivior in old rats.

  • Attenuating effect of drugs on an impairment of anticipatory activity in various animal models.

  • Systematic Analysis of Biological Timing

  • Development of animal models for Parkinson's desease and new drugs for this desease by the study of synaptic plasticity

  • 生物振動の発生とその多元的展開:生体機能の時間的組織化と複雑系

    科学研究費助成事業(北海道大学)  科学研究費助成事業(総合研究(B))

  • Molecular and Cellular Mechanisms of Biological Rhythms

  • Circadian clock function as a neuron-glia complex

  • シナップス可塑性の観点から見た体内時計への光入力

    科学研究費助成事業(早稲田大学)  科学研究費助成事業(特定領域研究(A))

  • 体内時計を視点とした情動行動機構解明

    科学研究費助成事業(早稲田大学)  科学研究費助成事業(特定領域研究(A))

  • 時計遺伝子発現による体内時計生理機構の解明

    科学研究費助成事業(早稲田大学)  科学研究費助成事業(特定領域研究(B))

  • 生体時計の分子機構

    科学研究費助成事業(神戸大学)  科学研究費助成事業(特定領域研究(B))

  • 体内時計階層構造システムから見た生体恒常性維持機構

    科学研究費助成事業(早稲田大学)  科学研究費助成事業(特定領域研究(A))

  • Molecular basis of circadian rhythms : function of clock genes

  • 時計遺伝子利用による分子時間薬理学研究の新展開

    科学研究費助成事業(早稲田大学)  科学研究費助成事業(萌芽的研究)

  • Research on molecular mechanism of sleep-wakefulness rhythm and development of new hypnotics

  • 光によるマスキング現象の解明と新規睡眠薬の開発

    科学研究費助成事業(早稲田大学)  科学研究費助成事業(萌芽研究)

  • Control mechanism of biological clocks in relation to mood, behavior and sleep disorders and Their treatments.

  • 体内時計の分子・生理機構

    科学研究費助成事業(早稲田大学)  科学研究費助成事業(基盤研究(C))

  • Studies of association of metabolic syndrome and circadian rhythms

  • 入眠のタイミングを司る物質の同定と新規入眠薬の開発

    科学研究費助成事業(早稲田大学)  科学研究費助成事業(萌芽研究)

  • Clock gene response against skelton type entrainment stimulation in mice

  • マウスの体内時計をリセットする食餌のパターン解析

    科学研究費助成事業(早稲田大学)  科学研究費助成事業(萌芽研究)

  • Role of breakfast in food-induced entrainment of mouse circadian clock

  • 時間栄養学の構築と応用研究

    科学研究費助成事業(早稲田大学)  科学研究費助成事業(挑戦的萌芽研究)

  • Mouse model searching for timing of ombination of feeding and exercise on health science

  • Establishment of chrono-exercise on mouse experiment

  • Development of novel in-vivo screening system for new psychotropic drags using robot technology

  • ニューロンにおけるシグナリング機構

    文部科学省 

▼display all

Presentations

  • 栄養指導に生かせる時間栄養学

    平成24年度メタボリックシンドローム予防のための健康セミナー 

    Presentation date: 2013.02

  • 時間遺伝子と食餌リズム

    味の素㈱東京支社 第19回「食と健康セミナー」 

    Presentation date: 2013.02

  • 「早寝早起き朝ごはん」のサイエンス

    農林水産省「平成24年度食育実践活動推進事業」シニア食育講座「すこやかシニアのための食生活講座」 

    Presentation date: 2013.02

  • 時計遺伝子と食事のリズム

    給食施設栄養管理研修会 

    Presentation date: 2013.01

  • Establishment of Shiftwork Model Mouse and Chrono-nutirional Approach for the Improvement of Shiftwork Induced Deficits

    Presentation date: 2012.12

  • Establishment of Shiftwork Model Mouse and Chrono-nutirional Approach for the Improvement of Shiftwork Induced Deficits

    Presentation date: 2012.12

  • Mouse circadian clock from cell to tissue

    The 1st Single-Cell Surveyor Symposium 

    Presentation date: 2012.11

  • マウス体内時計遺伝子をリセットする食餌内容の研究

    ㈶飯島記念食品科学振興財団 第25回学術講演会 

    Presentation date: 2012.11

  • 時間薬理学・時間栄養学の視点の生活習慣病

    平成24年度メタボリックシンドローム予防のための健康セミナー 

    Presentation date: 2012.11

  • 時間栄養学 −食の内容から食べ方まで−

    第20回日本植物油協会 植物油栄養懇話会 

    Presentation date: 2012.11

  • 体内時計と栄養

    第6回順天堂Kampo先端治療研究会 

    Presentation date: 2012.11

  • 時間栄養学〜免疫・炎症から代謝障害まで〜

    第5回基礎と臨床を結ぶ分子病態研究会 

    Presentation date: 2012.10

  • マウス絶食性脂肪肝はAChE阻害剤により増強する

    第127回日本薬理学会関東部会 

    Presentation date: 2012.10

  • 時間栄養学と肥満・代謝障害

    第33回日本肥満学会 

    Presentation date: 2012.10

  • 時間栄養学 〜早寝早起き朝ごはんを科学する〜

    平成24年度中原区栄養士・養護教諭等研修会 

    Presentation date: 2012.10

  • 食物アレルゲン摂取タイミングの違いがアレルギー症状に及ぼす影響

    第19回日本時間生物学会学術大会 

    Presentation date: 2012.09

  • シフトワークモデルのマウス体重増加及び行動リズムの乱れに与える効果

    第19回日本時間生物学会学術大会 

    Presentation date: 2012.09

  • 食餌による末梢時計同調の臓器間の差に関する研究

    第19回日本時間生物学会学術大会 

    Presentation date: 2012.09

  • マウス絶食性の脂肪肝における絶食の内容ならびにタイミングの効果

    第19回日本時間生物学会学術大会 

    Presentation date: 2012.09

  • 高脂肪食の摂食時刻・持続時間の体重や脂肪および末梢時計リズムへの影響

    第19回日本時間生物学会学術大会 

    Presentation date: 2012.09

  • 視床下部内側基底部において制限給餌に伴って発現量が変化する遺伝子群の検出

    第19回日本時間生物学会学術大会 

    Presentation date: 2012.09

  • 1週間リズム(Infradian rhythm)での光, 運動, 高脂肪食によるマウスの体重に与える影響

    第19回日本時間生物学会学術大会 

    Presentation date: 2012.09

  • 漬物がマウス肝臓体内時計遺伝子に与える影響

    第19回日本時間生物学会学術大会 

    Presentation date: 2012.09

  • 魚油摂取時のマウス肝臓の位相後退作用においてGPR120を介した新しい経路の確立

    第19回日本時間生物学会学術大会 

    Presentation date: 2012.09

  • GPR120遺伝子発現リズムとインスリン分泌に魚油が及ぼす影響

    第19回日本時間生物学会学術大会 

    Presentation date: 2012.09

  • マウス絶食性の脂肪肝における体内時計遺伝子の関与

    第19回日本時間生物学会学術大会 

    Presentation date: 2012.09

  • イン・ビボ イメージングを用いた老齢マウス末梢時計遺伝子発現の様態観測

    第19回日本時間生物学会学術大会 

    Presentation date: 2012.09

  • インビボとインビトロで調べた末梢時計の相違と特徴

    第19回日本時間生物学会学術大会 

    Presentation date: 2012.09

  • Chrononutrition -Interaction between circadian systems and feeding/nutrition-

    Lecture, UCLA 

    Presentation date: 2012.08

  • 生体リズムの応用と時間健康科学

    睡眠健康指導士 草津上級講座 睡眠学教育講座 

    Presentation date: 2012.08

  • 体内時計と栄養や健康・生活習慣病などとの関係について

    平成24年度公益社団法人宮崎県栄養士会生涯学習研修会 

    Presentation date: 2012.07

  • 「時計遺伝子と食事のリズム」

    第42回栄養学連続講義(夏期セミナー) 

    Presentation date: 2012.07

  • 「マウスを用いた時間栄養学視点からの塩分摂取と排泄機構の研究」

    第24回平成23年度助成研究発表会(公益財団法人ソルト・サイエンス研究財団) 

    Presentation date: 2012.07

  • 『喫煙と消化器系「マウス潰瘍性大腸炎に対する喫煙やニコチンの時間薬理学的作用機構解明」』

    第27回平成23年度助成研究発表会(公益財団法人喫煙科学研究財団) 

    Presentation date: 2012.07

  • 『生体リズム研究と時間治療の最前線「時間栄養学」』

    医療薬学フォーラム2012 第20回クリニカルファーマーシーシンポジウム 

    Presentation date: 2012.07

  • 『食と睡眠とリズム「時間栄養学の視点からより良いライフスタイルを考える」』

    日本睡眠学会第37回定期学術集会 

    Presentation date: 2012.06

  • 「時計遺伝子と時計栄養学」

    平成24年度生涯学習研修会 

    Presentation date: 2012.06

  • 「時間栄養学−細胞から動物」

    食品ニューティノロジー研究会(2012年6月例会) 

    Presentation date: 2012.06

  • 『時間栄養セミナー 長寿の秘訣を時計栄養から探る「時間栄養と食品成分の関係を知って健康維持に役立てる」』

    第17回国際食品素材/添加物展・会議 

    Presentation date: 2012.05

  • 「食事と時計遺伝子」

    第32回東京成長ホルモン成長因子セミナー 

    Presentation date: 2012.05

  • In vivo monitoring of peripheral circadian clocks in the mouse

    13th Biennial Meeting Society for Research on biological Rhythms 2012 

    Presentation date: 2012.05

  • Substitution of fish oil containing omega-3 fatty acid augmentsrestricted feeding-induced entrainment of the liver clockthrough GPR120 activation in PER2::LUCIFERASE mouse

    13th Biennial Meeting Society for Research on biological Rhythms 2012 

    Presentation date: 2012.05

  • Role of the Circadian Clock in Immediate- and Delayed-Type Skin Allergic Reaction in Mice

    13th Biennial Meeting Society for Research on biological Rhythms 2012 

    Presentation date: 2012.05

  • 「Clock変異マウスの食塩の嗜好性と含塩食物摂取が代謝機能に及ぼす影響」

    第66回日本栄養・食糧大会 

    Presentation date: 2012.05

  • 「理想的な1週間の生活リズムの追究−マウスの体重及び摂食量による評価−」

    第66回日本栄養・食糧大会 

    Presentation date: 2012.05

  • 「マウスの肝臓脂肪蓄積に対する低アミノ酸食と朝食・夕食の摂餌時刻の効果」

    第66回日本栄養・食糧大会 

    Presentation date: 2012.05

  • 「マウスやMEF細胞の体内時計周期に対する[6]-gingerolの作用」

    第66回日本栄養・食糧大会 

    Presentation date: 2012.05

  • 「魚油がマウス体内時計位相変動に与える新規パスウェイの構築」

    第66回日本栄養・食糧大会 

    Presentation date: 2012.05

  • 「時計遺伝子と食・栄養・エネルギー代謝」

    神戸市医師会学術講演会 

    Presentation date: 2012.05

  • 「インビボイメージングによるマスス給餌性末梢時計同調への給餌回数・間隔・量の要因分析」

    第89回日本生理学会大会 

    Presentation date: 2012.03

  • 「マウスにおける概日リズムと接触性過敏症の関係」

    第89回日本生理学会大会 

    Presentation date: 2012.03

  • 「マウスにおける水、スクロース、食塩入りスクロース飲水の選択行動に対する概日時計障害の影響

    第89回日本生理学会大会 

    Presentation date: 2012.03

  • 「栄養学・食事学の立場から−時間栄養と時間食事」

    第89回日本生理学会大会 

    Presentation date: 2012.03

  • 「体内時計と重金属毒性」

    第89回日本生理学会大会 

    Presentation date: 2012.03

  • 『食品の機能性を時間生物学的視点から考える「マウス体内時計と食・栄養の相互作用」』

    日本農芸化学会2012年度大会 

    Presentation date: 2012.03

  • 「食と時間栄養学」

    第1回機能性食品化学プロゼクトシンポジウム 

    Presentation date: 2012.03

  • 「体内時計と時間栄養(食事)学」

    関東信越国立病院管理栄養士協議会大会 

    Presentation date: 2012.03

  • 「体内時計と健康」

    創立30周年記念講演会(NHKエンジニアリングサービス) 

    Presentation date: 2012.03

  • 「マウス肝臓時計遺伝子リセットに魚油が与える影響」

    日本農芸化学会2012年度大会 

    Presentation date: 2012.03

  • 「時間栄養学から考える朝食の重要性−体内時計の仕組みと食事のリズム−」

    若い世代の食育フォーラム〜朝食の重要性について考えよう〜 

    Presentation date: 2012.02

  • 「睡眠・栄養・体内時計」(特別講演)

    第14回睡眠指導士初級養成講座 

    Presentation date: 2012.01

  • 「時間栄養学と生活習慣病の予防」(特別講演)

    平成23年度いばらき食育推進大会 

    Presentation date: 2012.01

  • Chrononutrition and chronodietetics

    Presentation date: 2011.12

  • 『生体リズムと創薬「時間薬理学の到達点」』

    第32回日本臨床薬理学会年会 

    Presentation date: 2011.12

  • 「輝度と色調を変えたテレビの視聴が唾液中のメラトニンとコルチゾールに及ぼす影響」

    第18回日本時間生物学会学術大会 

    Presentation date: 2011.11

  • 「アデノシン・P2Y受容体の体内時計機構における役割解明」

    第18回日本時間生物学会学術大会 

    Presentation date: 2011.11

  • 「カフェインが体内時計周期に与える影響とその機序解明」

    第18回日本時間生物学会学術大会 

    Presentation date: 2011.11

  • 「リモネンとレモン果汁の同時摂取による時計遺伝子発現量への影響」

    第18回日本時間生物学会学術大会 

    Presentation date: 2011.11

  • 「リモネンやリナロールの匂い刺激がマウスの時計遺伝子発現に及ぼす影響」

    第18回日本時間生物学会学術大会 

    Presentation date: 2011.11

  • 「魚油がマウス体内時計位相後退作用に及ぼすメカニズムの解明」

    第18回日本時間生物学会学術大会 

    Presentation date: 2011.11

  • 「時間依存的な摂餌パターンの変化に伴う脂肪肝の発生メカニズム解明」

    第18回日本時間生物学会学術大会 

    Presentation date: 2011.11

  • 「時計遺伝子と食事のリズム−時間栄養学−」

    第33回日本臨床栄養学会総会 

    Presentation date: 2011.10

  • 『精神神経疾患と生体リズム「生体リズムに対する抗精神病薬の影響」』(シンポジウム)

    第41回日本神経精神薬理学会 

    Presentation date: 2011.10

  • 『食事と体内時計「体内時計の仕組みと食事のリズム」』(特別講演)

    群馬県栄養士会研修会 

    Presentation date: 2011.10

  • Potential therapeutic and diagnostic targets for circadian rhythm sleep disorders

    Worldsleep 2011 

    Presentation date: 2011.10

  • 「薬の効き方・使い方を学ぶ薬理学のいろは」

    EWE三月会講演会 

    Presentation date: 2011.09

  • 「時間栄養学の視点から」

    第87回日本栄養・食糧学会関東支部大会シンポジウム 

    Presentation date: 2011.07

  • 「各澱粉含有食がマウス肝臓に及ぼす体内時計リセット効果」

    第65回日本栄養・食糧学会大会 

    Presentation date: 2011.05

  • 「マウス組織において魚油がスギ花粉のアレルギー症状に与える影響」

    第65回日本栄養・食糧学会大会 

    Presentation date: 2011.05

  • Refeeding After Fasting Elicits Insulin-dependent Regulation Of Per2 and Rev-erba with Shifts in liver clock

    3rd World Congress of Chronobiology 

    Presentation date: 2011.05

  • About the relation nature of the day-and-night difference of a blood glucose level and insulin rise of a mouse

    3rd World Congress of Chronobiology 

    Presentation date: 2011.05

  • Role of food components in entrainment of mouse liver clock

    3rd World Congress of Chronobiology 

    Presentation date: 2011.05

  • Food entrainment of peripheral clocks evaluated by in vivo imaging

    3rd World Congress of Chronobiology 

    Presentation date: 2011.05

  • Chrono-nutrition and chrono-dietetics

    XI Latin America Symposium of Chronobiology 

    Presentation date: 2011.05

  • 「食事と体内時計」

    AACC International 日本支部例会 

    Presentation date: 2011.04

  • 低アミノ酸食による脂肪肝と体内時計の関係

    第6回環境生理学プレコングレス 

    Presentation date: 2011.03

  • 「時間栄養学 〜体内時計と食事のリズム〜」

    多摩区栄養士研修会 

    Presentation date: 2011.02

  • 『くらしの中の糖尿病「生活リズムと体内時計」』

    第9回糖尿病市民セミナー・東京 

    Presentation date: 2011.02

  • 「時計遺伝子と食餌リズム」

    第13回脂質・栄養シンポジウム 

    Presentation date: 2011.01

  • 現代生活の「時計」と「休息」・「からだとこころの健康の実践:時間栄養学の考え方」

    早稲田大学教育総合研究所 教育最前線講演会シリーズXI 

    Presentation date: 2010.12

  • インビボ・イメージングによるマウス末梢時計測定法確率と末梢時計の中枢制御

    第17回日本時間生物学会学術大会 

    Presentation date: 2010.11

  • マウスにおけるスギ花粉監査時間の変化がアレルギー反応に与える影響

    第17回日本時間生物学会学術大会 

    Presentation date: 2010.11

  • マウス末梢時計の給餌性リズム同調におけるスターチの影響

    第17回日本時間生物学会学術大会 

    Presentation date: 2010.11

  • アドレナリン受容体の刺激がPer2リズム周期に与える影響

    第17回日本時間生物学会学術大会 

    Presentation date: 2010.11

  • 麻酔薬が時計遺伝子発現リズムに与える影響

    第17回日本時間生物学会学術大会 

    Presentation date: 2010.11

  • マウス体内時計と食・栄養の相互作用

    第17回日本時間生物学会学術大会 

    Presentation date: 2010.11

  • 子どもの脳の発達に不可欠な睡眠とそのリズム

    第2回神戸市民講座「赤ちゃんから、お母さんと社会へのメッセージ」 

    Presentation date: 2010.10

  • Development of Experimental Setup to Create Novel Mental Disorder Model Rats using Small Mobile Robot

    IROS 2010 

    Presentation date: 2010.10

  • 体内時計と食・栄養科学

    第57回日本食品科学工学会 

    Presentation date: 2010.09

  • リズム障害と生活習慣病

    日本睡眠学会第35回定期学術集会 

    Presentation date: 2010.07

  • Fish oil affects clock gene and clock-controlled metabolism-related gene expression in the mouse liver

    WorldPharma2010 

    Presentation date: 2010.07

  • Chinese traditional medicine on clock- and metabolism-related gene expression in mice

    WorldPharma2010 

    Presentation date: 2010.07

  • インビボ・イメージングによるマウス体内時計測定法と視交叉上核破壊による末梢時計への影響

    BMB2010 日本分子生物学会・日本生化学会合同大会 

    Presentation date: 2010.07

  • エイジングとアンチエイジングに対する体内時計の関わり

    私立大学戦略的基盤形成研究支援事業 第2回学術講演会「エイジングに伴う生体機能変化とアンチエイジング戦略」 近畿大学アンチエイジングセンター 

    Presentation date: 2010.06

  • 食餌の成分・タイミングの相違がマウスの体重・体脂肪量ならびに遺伝子発現に及ぼす影響

    第64回日本栄養・食糧学会大会 

    Presentation date: 2010.05

  • 十全大補湯がマウスの時計遺伝子及び代謝関連遺伝子に与える効果

    第64回日本栄養・食糧学会大会 

    Presentation date: 2010.05

  • 高GI食がもたらすマウス肝臓時計遺伝子発現への効果

    第64回日本栄養・食糧学会大会 

    Presentation date: 2010.05

  • Insulin-dependent Per2 gene expression is the first entrainment signal to the peripheral circadian clocks by food or glucose.

    12th Biennial Meeting Society for Research on Biological Rhythms 

    Presentation date: 2010.05

  • Variance of feeding time influences body weight, body fat, and gene expressions in mice.

    12th Biennial Meeting Society for Research on Biological Rhythms 

    Presentation date: 2010.05

  • Attenuated food anticipatory activity and abnormal circadian locomotor rhythms in Rgs16 knockdown mice.

    12th Biennial Meeting Society for Research on Biological Rhythms 

    Presentation date: 2010.05

  • A relationship between circadian rhythm and pathogenesis of atopic dermatitis-like skin lesions in mice.

    The 87th Annual Meeting of the Physiological Society of Japan 

    Presentation date: 2010.05

  • The effect of exercise on the phase of circadian rhythm and metabolic-related genes in mice.

    The 87th Annual Meeting of the Physiological Society of Japan 

    Presentation date: 2010.05

  • Research of mouse circadian rhythm mased on chrononutrition approach

    The 87th Annual Meeting of the Physiological Society of Japan 

    Presentation date: 2010.05

  • 時計遺伝子と時間栄養学

    平成22年度茨城県栄養士会特別講演会 

    Presentation date: 2010.05

  • 体内時計研究の基礎から応用

    山梨大学?早稲田大学連携事業セミナー 

    Presentation date: 2010.04

  • Effect of chinese herb Jyuzen-taiho-to clock gene and metabolism related genes in the mouse liver.

    The 83rd Annual Meeting of The Japanese Pharmacological Society 

    Presentation date: 2010.03

  • Chronopharmacological and molecular studies of anti-diuretic drugs on Per2 mutant mice.

    The 83rd Annual Meeting of The Japanese Pharmacological Society 

    Presentation date: 2010.03

  • Progress in chrononutrition

    The 83rd Annual Meeting of The Japanese Pharmacological Society 

    Presentation date: 2010.03

  • 体内時計オーバービュー

    日本薬学会第130年会シンポジウム「体内時計を利用した創薬から治療戦略まで」 

    Presentation date: 2010.03

  • バランスのとれた高GI食がマウス肝臓時計遺伝子に与える影響

    日本農芸化学会2010年度大会 

    Presentation date: 2010.03

  • Nutrition and Circadian Clock System

    Presentation date: 2010.03

  • 体内時計とメタボリックシンドローム

    朝日生命成人病研究所 126回分泌セミナー 

    Presentation date: 2010.02

  • リズム障害の基礎と応用

    第5回関東睡眠談話会特別講演 

    Presentation date: 2010.02

  • 体内時計と食・栄養

    ATI(新世代研究所) 時を計る研究会 

    Presentation date: 2009.12

  • 体内時計と食・栄養の関わり

    第3回ニュートリズム検討会 

    Presentation date: 2009.09

  • Critical role of dorsomedial hypothalamus (DMH) in light-induced but not fiiding-induced entrainment of liver circadian rhythm in Per2::luc knock-in mouse.

    XI. Congress of the European Biological Rhythms Society 

    Presentation date: 2009.08

  • Balanced nutrient foods with high glycemic index but not sugar alone is suitable entraining signals of mouse liver clock.

    XI. Congress of the European Biological Rhythms Society 

    Presentation date: 2009.08

  • Neural and peripheral mechanisms

    XI. Congress of the European Biological Rhythms Society 

    Presentation date: 2009.08

  • Symposium 8. Interplay between circadian clocks and metabolism

    XI. Congress of the European Biological Rhythms Society 

    Presentation date: 2009.08

  • 給餌性体内時計リズム形成における交感神経作用薬の作用

    第63回日本栄養・食糧学会大会 

    Presentation date: 2009.05

  • 給餌によるマウス肝臓時計遺伝子発現リズムに対する餌の量とタイミングの効果

    第63回日本栄養・食糧学会大会 

    Presentation date: 2009.05

  • 食事の量やタイミングの相違がマウスの体重・脂肪増加に及ぼす影響

    第63回日本栄養・食糧学会大会 

    Presentation date: 2009.05

  • 体内時計が生命科学研究にどのように役立つのか?

    学術シンポジウム:“山梨大学・早稲田大学:国私立大学間連携による医学・理工学に精通した先端生命科学分野の国際的研究者の育成” 

    Presentation date: 2009.02

  • 体内時計と栄養の観点から見た食育〜親が変われば子供が変われる生活リズム〜

    東京都多摩立川保健所 食育研究会 

    Presentation date: 2009.01

  • 子供の脳の発達に不可欠な睡眠とそのリズム

    (財)パブリックヘルスリサーチセンター 市民講座「赤ちゃんからお母さんと社会へのメッセージ」 

    Presentation date: 2008.12

  • 食事や食品成分による時計遺伝子発現リズムの制御機構

    (社)日本食品科学工学会関東支部 平成20年度大会 

    Presentation date: 2008.11

  • 早寝早起き朝ごはんのサイエンス

    早稲田電気工学会 三月会9月定例会 

    Presentation date: 2008.09

  • ニコチン投与性の新規リズム形成と人への応用

    喫煙科学研究財団 第23回助成研究発表会 

    Presentation date: 2008.07

  • 分子モデルによる哺乳類の概日リズムのロバスト性解析

    早稲田電気工学会 三月会9月定例会 

    Presentation date: 2008.05

  • 食事と体内時計

    (社)島根県栄養士会 生涯学習研修会 

    Presentation date: 2008.05

  • 食事と体内時計

    千葉県病院栄養士協議会 平成20年度研修会 

    Presentation date: 2008.04

  • Nutritional analysis of food-entrained oscillation in the mouse liver.

    The 85th Annual Meeting of the Physiological Society of Japan 

    Presentation date: 2008.03

  • Effect of tobacco smoking on respiratory response in OVA-induced asthma model mouse.

    The 85th Annual Meeting of the Physiological Society of Japan 

    Presentation date: 2008.03

  • The neural mechanisms underlying the food-associated rhythms in mice.

    The 85th Annual Meeting of the Physiological Society of Japan 

    Presentation date: 2008.03

  • Effects of PPARα ligand on circadian rhythms and EEG delta power in mice.

    The 85th Annual Meeting of the Physiological Society of Japan 

    Presentation date: 2008.03

  • Effect of glycine on light-induced phase shift and c-fos expression in the suprachiasmatic nucleus and retina of mice.

    The 85th Annual Meeting of the Physiological Society of Japan 

    Presentation date: 2008.03

  • Circadian and ultradian change of body temperature rhythm in ovarectmized menopausal model mice

    The 85th Annual Meeting of the Physiological Society of Japan 

    Presentation date: 2008.03

  • A role of E and/or M oscillator in clock gene expression rhythm of mouse liver

    The 85th Annual Meeting of the Physiological Society of Japan 

    Presentation date: 2008.03

  • Meaning of breakfast on food-entrained oscillation

    The 85th Annual Meeting of the Physiological Society of Japan 

    Presentation date: 2008.03

  • システム生命科学による医・理・工学の先端研究

    東京女子医科大学・早稲田大学ジョイントシンポジウム「医・理・工融合研究の新しい連携のかたち」 

    Presentation date: 2008.03

  • 給餌性リズム形成における朝食の意味

    第85回日本生理学会大会シンポジウム 

    Presentation date: 2008.03

  • 食事と体内時計

    (社)広島県栄養士会 平成20年度研修会 

    Presentation date: 2008.02

  • 体内時計と食と栄養

    講演会「早寝早起き朝ごはん」で心と体の活力を高めよう! 

    Presentation date: 2008.01

  • 時間生物学から見た向精神薬の薬理作用

    日本睡眠学会第32回定期学術集会・第14回日本時間生物学会学術大会 合同大会 

    Presentation date: 2007.11

  • 朝食と時間生物学

    APACPHケロッグランチセッション(日本ケロッグ) 

    Presentation date: 2007.11

  • 体内時計を活用した健康法−マウスからヒトまで

    第8回日本健康支援学会年次学術集会 

    Presentation date: 2007.02

  • 時計遺伝子の概日リズムに対する光と食事の作用

    講演会「早寝早起き朝ごはん」はなぜ健康によいのか? 

    Presentation date: 2007.01

  • 制限給餌誘発性リズムの振動体としての視床下部背内側核の役割

    第57回日本薬理学会北部会 

    Presentation date: 2006.09

  • 外から見た薬学

    平成18年度九大薬友会関東支部総会 

    Presentation date: 2006.05

  • 高脂血症・肥満における体内時計の役割

    第79回日本薬理学会年会シンポジウム「体内時計遺伝子が持つ多彩な機能—疾病発現から治療薬まで」 

    Presentation date: 2006.03

  • 生体リズムに対する運動の効果

    第83回日本生理学会大会 

    Presentation date: 2006.03

  • 高脂肪食におけるICR系クロックミュータントマウス脂肪組織への影響

    第83回日本生理学会大会 

    Presentation date: 2006.03

  • 発達期の光環境条件によるClock遺伝子一部欠損マウスのDSPS(delayed sleep phase syndrome)発現・特別講演

    第83回日本生理学会大会サテライト企画 第2回環境生理学プレコングレス 

    Presentation date: 2006.03

  • 体内時計変異マウスの子育て行動

    早稲田大学人間総合研究センター学術フロンティア「脳機能の性差形成機構」公開シンポジウム 

    Presentation date: 2006.01

  • 体内時計研究 −時刻認知から薬物治療−

    脳の医学・生物学研究会 第39回研究会 

    Presentation date: 2005.07

  • 老化と睡眠・リズム

    早稲田大学人間総合研究センター「生活の質」土曜講座 高齢者のこころとからだ 第243回 

    Presentation date: 2005.01

  • 体内時計の分子から生理・薬理機能解明へ

    第76回日本薬理学会第80回日本生理学会(共同開催)シンポジウム 

    Presentation date: 2003.03

  • マウス視交叉上核スライス標本の発火リズムに対するグレリンの効果

    第80回日本生理学会年会 

    Presentation date: 2003.03

  • 時間薬理をめざした体内時計研究

    第23回日本臨床薬理学会イブニングセミナー「第3回 時間薬理・治療を考える会」 

    Presentation date: 2002.12

  • ストレプトゾトシン投与マウスの肝臓におけるPER2発現リズム

    第9回日本時間生物学会学術大会 

    Presentation date: 2002.11

  • 概日リズム発振におけるClock遺伝子変異の影響

    第9回日本時間生物学会学術大会 

    Presentation date: 2002.11

  • 感情障害治療薬と体内時計遺伝子発現

    第9回日本時間生物学会学術大会シンポジウム 

    Presentation date: 2002.11

  • 遺伝子発現と治療薬からみた体内時計研究

    第9回日本時間生物学会学術大会シンポジウム 

    Presentation date: 2002.11

  • オレキシンおよびオレキシン受容体発現に対する制限給餌の影響

    第8回日本時間生物学会学術大会 

    Presentation date: 2001.11

  • リズム同調の分子・薬理機構

    日本疾患モデル学会第18回総会 シンポジウム「時計遺伝子とその異常」 

    Presentation date: 2001.11

  • 生体時計リズム

    第24回日本神経科学・第44回日本神経化学合同大会シンポジウム「リズムとゆらぎの神経科学」 

    Presentation date: 2001.09

  • 記憶・学習関連行動の神経科学的基盤

    第74回日本薬理学会年会 

    Presentation date: 2001.03

  • 生体リズム階層性と同調機構で

    第6回慢性疲労症候群(CFS)研究会 

    Presentation date: 2001.02

  • 体内時計階層構造システムから見た生体恒常性維持

    [総合脳]公募班研究成果報告会 

    Presentation date: 2000.12

  • Non-photic entrainment of Per gene expression in mouse brain and liver.

    U.S.-Japan Symposium on Molecular Mechanism for Circadian Clocks: Functions of Clock Genes. 

    Presentation date: 2000.12

  • 脳のリズム

    シンポジウム“ニューロシグナリングから知識工学への展開” 

    Presentation date: 2000.11

  • 短時間作動型睡眠導入薬ブロチゾラムのハムスターの輪回し行動リズムとSNC内での Period mRNA の発現に対する効果

    第7回日本時間生物学会学術大会 

    Presentation date: 2000.11

  • ラット視交叉上核におけるLTPの情報伝達系とメラトニンによる調節

    第7回日本時間生物学会学術大会 

    Presentation date: 2000.11

  • 時計遺伝子発現からみた給餌性リズム形成におけるグルコースの関与

    第7回日本時間生物学会学術大会 

    Presentation date: 2000.11

  • 時刻学習と体内時計機構

    第7回日本時間生物学会学術大会 

    Presentation date: 2000.11

  • A combined study of TRACT-TRACING and electrophysiological mapping in area te and area 36 of macaque monkeys

    SOCIETY FOR NEUROSCIENCE, 30TH ANNUAL MEETING 

    Presentation date: 2000.11

  • 体内時計をリセットする光と光以外の同調因子の作用機構

    第73回日本生化学会 

    Presentation date: 2000.10

  • GRP受容体遺伝子欠損マウスの体内時計の光同調低下

    第23回日本神経科学大会・第10回日本神経回路学会合同大会 

    Presentation date: 2000.09

  • PACAPのマウス体内時計の同調機構に及ぼす影響

    第23回日本神経科学大会・第10回日本神経回路学会合同大会 

    Presentation date: 2000.09

  • マウス視交叉上核におけるPeriod遺伝子発現リズムの胎生期・生後発達

    第23回日本神経科学大会・第10回日本神経回路学会合同大会 

    Presentation date: 2000.09

  • マウスにおける恒明状態での行動リズム異常と時計遺伝子の発現リズムについて

    第23回日本神経科学大会・第10回日本神経回路学会合同大会 

    Presentation date: 2000.09

  • Effect of PACAP on Entrainment of Mice Circadian Rhythm.

    Neuroscience Research-Yokohama 

    Presentation date: 2000.09

  • Differential Daily Expression of Per1 and per2 mRNA in the Suprachiasmatic Nucleus of Fetal and Early Postnatal Mice

    Neuroscience Research-Yokohama 

    Presentation date: 2000.09

  • Behavioral free-running and rhythmic expression of the clock genes under constant lighting conditions in mice.

    Neuroscience Research-Yokohama 

    Presentation date: 2000.09

  • マウスSCNにおけるmCry1 mRNAの発現と行動リズムについて

    第6回日本時間生物学会学術大会 

    Presentation date: 1999.11

  • マウス小脳顆粒細胞培養系におけるmPer遺伝子発現調節

    第6回日本時間生物学会学術大会 

    Presentation date: 1999.11

  • ストレスによるmPer遺伝子発現調節

    第6回日本時間生物学会学術大会 

    Presentation date: 1999.11

  • マウスにおける周期的制限給餌による肝臓Period mRNA発現への影響について

    第6回日本時間生物学会学術大会 

    Presentation date: 1999.11

  • 制限給餌におけるSCN非依存的給餌性リズムと 遺伝子の発現様式の解析

    第6回日本時間生物学会学術大会 

    Presentation date: 1999.11

  • 光刺激によってSCN内で発現増強されるPeriod mRNAに対するCaMキナーゼII

    第6回日本時間生物学会学術大会 

    Presentation date: 1999.11

  • 光刺激による概日リズムの位相変化に伴ったハムスター Period mRNAの発現分布

    第6回日本時間生物学会学術大会 

    Presentation date: 1999.11

  • ラット、マウスの老齢動物における Period mRNA発現パターンの加齢による変化

    第6回日本時間生物学会学術大会 

    Presentation date: 1999.11

  • メタンフェタミンの薬理作用とmPer遺伝子発現

    第6回日本時間生物学会学術大会 

    Presentation date: 1999.11

  • ラット視交叉上核へのPer antisenseoligonucleotide投与

    第6回日本時間生物学会学術大会 

    Presentation date: 1999.11

  • 薬物による概日リズムの位相変化に 伴ったげっ歯類mRNAの発現動態

    第5回日本時間生物学会学術大会 

    Presentation date: 1998.11

  • 光刺激で誘導される哺乳類per遺伝子ホモログの相互作用解析

    第5回日本時間生物学会学術大会 

    Presentation date: 1998.11

  • マウス小脳に発現するmPer遺伝子に対する抑制薬の作用

    第5回日本時間生物学会学術大会 

    Presentation date: 1998.11

  • 周期的制限給餌性リズムにおけるmPer遺伝子群の関与

    第5回日本時間生物学会学術大会 

    Presentation date: 1998.11

  • メラトニン、メチルコバラミンの飲水投与による明暗サイクルの連続アドヴァンスに対する再同調促進作用 −行動リズムならびにmPer1 mRNA発現による評価−

    第5回日本時間生物学会学術大会 

    Presentation date: 1998.11

  • mPer1-3 antisenseoligonucleotideのマウスサーカディアンリズムに対する作用

    第5回日本時間生物学会学術大会 

    Presentation date: 1998.11

  • Triazolamにより誘発されるハムスターの輪回し行動リズムの 位相前進作用に対するMKC-242の効果

    第5回日本時間生物学会学術大会 

    Presentation date: 1998.11

  • Ryanodine receptor type3(RyR3) ノックアウトマウスの行動リズム解析

    第5回日本時間生物学会学術大会 

    Presentation date: 1998.11

  • ラットの行動リズムにおけるrat per1 antisense oligonucleotide投与の影響

    第5回日本時間生物学会学術大会 

    Presentation date: 1998.11

  • メタンフェタミン惹起性予知行動リズムと mPer遺伝子

    第5回日本時間生物学会学術大会 

    Presentation date: 1998.11

  • グルタミン酸による視交叉上核神経活動の位相変化に対するmPer1アンチセンスオリゴヌクレオチドの効果

    第5回日本時間生物学会学術大会 

    Presentation date: 1998.11

  • 糖尿病モデルOtsuka Long Evans Tokushima Fatty(OLETF)ラットの体内時計機能異常

    第5回日本時間生物学会学術大会 

    Presentation date: 1998.11

  • 恒暗条件化のハムスター輪回しリズムに対するR(+)8-OHDPATのarousal型位相変化

    第4回日本時間生物学会学術大会 

    Presentation date: 1997.11

  • 体内時計におけるNMDA受容体サブタイプの役割

    第4回日本時間生物学会学術大会 

    Presentation date: 1997.11

  • NMDA受容体サブタイプノックアウト動物に対する制限給餌同調の効果

    第4回日本時間生物学会学術大会 

    Presentation date: 1997.11

  • カルシウムイメージング法を用いた視交叉上核NMDA受容体の機能解析

    第4回日本時間生物学会学術大会 

    Presentation date: 1997.11

  • アストログリア特異的に発現するGlial Fibrillary Acidic Protein(GFAP)の概日リズムにおける役割について

    第4回日本時間生物学会学術大会 

    Presentation date: 1997.11

  • ラット新生仔概日リズム形成に対するD1-DopamineとMelatoninの同調因子としての影響

    第4回日本時間生物学会学術大会 

    Presentation date: 1997.11

  • サーカディアンリズムの研究:創薬から治療薬へ

    第4回日本時間生物学会学術大会シンポジウム 

    Presentation date: 1997.11

  • Glial Fibrillary Acidic Protein(GFAP)遺伝子欠損マウスの行動リズム研究

    第3回日本時間生物学会学術大会 

    Presentation date: 1996.11

  • 老化促進マウス(SMAP-8)の自発運動のサーカディアンリズムに対する薬物の作用

    第3回日本時間生物学会学術大会 

    Presentation date: 1996.11

  • 視交叉上核における2つの長期増強現象

    第3回日本時間生物学会学術大会 

    Presentation date: 1996.11

  • 老齢ラットにおけるSCNからのVIP遊離に対するメラトニンの影響

    第3回日本時間生物学会学術大会 

    Presentation date: 1996.11

  • 視交叉上核における長期増強現象

    第2回日本時間生物学会学術大会 

    Presentation date: 1995.11

  • 視神経刺激及び高濃度KClによる視交叉上核からのVIP遊離と加齢による影響

    第2回日本時間生物学会学術大会 

    Presentation date: 1995.11

  • ラット視交叉上核への光入力に対するメタンフェタミンの抑制作用

    日本時間生物学会第1回設立記念学術集会 

    Presentation date: 1994.10

  • 視交叉上核(SCN)への光情報入力に於ける一酸化窒素(NO)合成の関与について

    日本時間生物学会第1回設立記念学術集会 

    Presentation date: 1994.10

  • 光同調入力経路の網膜−視床下部路におけるGABA神経の役割

    日本時間生物学会第1回設立記念学術集会 

    Presentation date: 1994.10

  • 神経伝達物質と老化

    日本時間生物学会第1回設立記念学術集会 

    Presentation date: 1994.10

  • ラット視交叉上核のVIP遊離における興奮性神経入力の役割

    第10回生物リズム研究会 

    Presentation date: 1993.11

  • ラット視交叉上核神経活動のサーカディアンリズムに対するNMDAの作用−NOの関与について−

    第10回生物リズム研究会 

    Presentation date: 1993.11

  • 制限給餌によるラットの予知行動−老化、NMDA受容体拮抗薬の影響

    第9回生物リズム研究会 

    Presentation date: 1992.09

  • ラット網膜視床下部路の伝達物質:培養神経組織を用いた薬物位相反応曲線による解析

    第9回生物リズム研究会・臨床時間生物学研究会・合同シンポジウム 

    Presentation date: 1992.09

  • NMDA受容体と時刻認知学習

    総合研究大学院大学研究会発表原稿集、 生体の時間秩序発現機構 

  • 生体の時計機構とその異常に関する研究

    ヒューマンサイエンス基礎研究事業、 官民共同プロジェクト研究報告 

  • Fear conditioning behavior in GFAP mutant mice.

    Japan Journal of Pharmacology 

  • Drinking administration of melatonin normalizes an impairment of re-entrainment to new light-dark cycle in senescence accelerated mouse.

    Japan Journal of Pharmacology 

  • Occurrence of sensitization to methamphetamine-induced disorganization of locomotor activity rhythm in rats.

    Japan Journal of Pharmacology 

  • Effect of foot-shock stress on light-induced entrainment of circadian wheel-running rhythm in hamsters.

    Japan Journal of Pharmacology 

  • Potentiating action of MKC-242, a selective 5-HT1A receptor agonist, on photic entrainment of circadian activity rhythm in rats.

    Japan Journal of Pharmacology 

  • Involvement of presynaptic 5-HT1A receptor agonistic action in the MKC-242-induced potentiation of photic entrainment in circadian rhythm.

    Japan Journal of Pharmacology 

  • SAMP8のサーカディアンリズムの特徴とメラトニン飲水投与の効果

    老化促進モデルマウス研究協議会抄録集 

  • k-opioid receptor agonistのサーカディアンリズム光同調に対する抑制作用

    日本薬理学雑誌 

  • 活動促進状態時のメラトニンの抑制作用

    日本薬理学雑誌 

  • 体内時計の光同調に対する新規セロトニン1A受容体アゴニストMKC242の増強作用

    日本薬理学雑誌 

  • Role of presynaptic serotonin receptors on rodent circadian rhythms, studies from hamsters to mutant mice.

    Japan/USA conference on Molecular Chronobiology 

  • 遺伝子アンチセンスによる マウス概日性活動リズムの位相変化の抑制

    第28回日本神経精神薬理学会プログラム集 

  • 海馬や小脳に発現する体内時計遺伝子mperに対する 覚醒薬ならびに抑制薬の作用

    第28回日本神経精神薬理学会プログラム集 

  • 哺乳類体内時計の光同調経路におけるカルモヂュリンキナーゼIIの役割について

    第21回日本神経科学プログラム集 

  • mper1遺伝子とmper2遺伝子のアンチセンスは光によるマウスの概日性運動リズムの位相変化を抑制する

    第21回日本神経科学プログラム集 

  • 音恐怖条件づけ関連神経核におけるCa2+応答

    第21回日本神経科学プログラム集 

  • NMDAレセプターノックアウトマウスを用いた恐怖条件学習と扁桃体におけるCaMKIIの燐酸化

    第21回日本神経科学プログラム集 

  • Fear conditioning behavior in GFAP mutant mice.

    Japan Journal of Pharmacology 

  • Drinking administration of melatonin normalizes an impairment of re-entrainment to new light-dark cycle in senescence accelerated mouse.

    Japan Journal of Pharmacology 

  • Occurrence of sensitization to methamphetamine-induced disorganization of locomotor activity rhythm in rats.

    Japan Journal of Pharmacology 

  • Effect of foot-shock stress on light-induced entrainment of circadian wheel-running rhythm in hamsters.

    Japan Journal of Pharmacology 

  • Potentiating action of MKC-242, a selective 5-HT1A receptor agonist, on photic entrainment of circadian activity rhythm.

    Japan Journal of Pharmacology 

  • Involvement of presynaptic 5-HT1A receptor agonistic action in the MKC-242-induced potentiation of photic entrainment in circadian rhythm.

    Japan Journal of Pharmacology 

  • ラット行動リズムにおけるrPer1 antisenseoligonucleotide投与の影響

    第22回日本神経科学プログラム集 

  • マウス小脳顆粒細胞培養系におけるmPer遺伝子発現の調節

    第22回日本神経科学プログラム集 

  • 体内時計リセットの学習/記憶から見た側面

    第22回日本神経科学プログラム集 

  • 体内時計の光同調シグナル系に対するAMPA受容体活性促進薬アニラセタムの増強効果

    第22回日本神経科学プログラム集 

  • NMDAR2欠損による扁桃体ニューロンのアゴニスト感受性の変化

    第22回日本神経科学プログラム集 

  • 体内時計遺伝子と高次脳機能

    日本組織細胞化学会第40回記念総会・学術集会 

  • 社会的ストレス誘発性マウス高体温に対するDR4004の効果

    第72回日本薬理学会年会 

  • 室傍核や扁桃体の時計遺伝子mPer1,mPer2 mRNAは ストレス負荷により誘発される

    第72回日本薬理学会年会 

  • げっ歯類体内時計の光同調に対するアニラセタムの増強作用

    第72回日本薬理学会年会 

  • NMDA受容体遺伝子欠損マウスにおける音恐怖条件づけ学習とCaMKIIの活性化

    第72回日本薬理学会年会 

  • 生体リズムと病気:創薬の視点に立った生体リズム

    日本薬学会第120年会 

  • グルタミン酸受容体を介した光による時計遺伝子ピリオドの発現上昇と体内時計のリセットについて

    第73回日本薬理学会年会 

▼display all

Specific Research

  • 脳ドパミン神経を制御する腸内細菌の仕組解明

    2020  

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    マウスに普通食と高脂肪食を選択的に与えると、マウスは好んで高脂肪食を食べるようになる。いわゆる依存的な食欲の亢進である。高脂肪食に対する依存状態が観察されたマウスの線条体や側坐核のドパミン神経を調べると、ドパミン量が有意に低下することが分かった。一方、申請者は高脂肪食を与えたマウスの腸内細菌は多様性の低下など、dysbiosis (腸内菌共生バランス失調)が起こること事を見出した。次に、このように腸内細菌の不調が、脳のドパミン量の低下に関連するのではないかと考え、dysbiosisを改善させるべく、水溶性食物繊維を同時に与えた。イヌリンを高脂肪食に添加することで、ドパミンの低下が改善された。

  • 社会的時差ボケマウスモデルの作成とヒトへの応用研究

    2017  

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    ヒトの場合、平日は起床が早いが休日は起床が遅く、睡眠負債を解消しようとしている。これに合わせて、マウスでは2日間(ヒトの土日)は明暗周期の位相を3あるいは6時間遅くし、次の5日間(ヒトの月曜から金曜)は明暗環境を元の位相に戻した。その結果、活動リズムの位相は2日間の明暗後退で大きく後退し、次の位相前進刺激に対しては反応が弱く、十分に元の位置に戻すことはできなかった。また、中枢の視交叉上核の時計はより速やかに位相が戻る傾向が見られたが、末梢時計の時計遺伝子発現リズムの位相は遅れ、乖離状態が見られた。今回の研究により、ヒトに見られる特徴を有した社会的時差ボケモデルマウスが完成した。

  • 週間リズムの視点の運動科学・栄養科学研究

    2014  

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    体内時計の研究成果の出口として、日内リズムと食・栄養の関係がわかり、メタボリックシンドロームの予防に「時間栄養学」の研究が進んできた。そこで、マウスを用いて週間リズムに焦点を当てた研究を展開した。先進国では、weekday(平日) とweekend(週末)の過ごし方が異なり、平日は短睡眠になり、週末は長睡眠となり、社会的時差ボケ状態になっている。マウスを用いて週間リズムが、生体のエネルギー代謝に、どのように影響を及ぼすかについて調べた結果、平日と週末が6時間異なるモデルでは、夜型化が進み、体内時計が遅れ、かつ肥満になった。現在3時間異なるモデルの作成を行ったり、これを改善する食パターンを提案する予定である。

  • 食欲と脂肪蓄積の時間栄養学的アプローチ研究

    2013  

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    脂肪代謝の時間栄養学的研究ということで、まず、脂肪の血中動態を指標に、時間栄養学的な研究を、次に特定の脂肪酸を含有する脂肪が体内時計の位相に形成に影響する可能性について調べた。脂質としては、DHA,EPAの含量が高い魚油、脂肪の吸収が速やかで燃焼効果が高いとされている中鎖脂肪酸の脂質、およびラードをそれぞれ0.1ml経口投与した。投与時刻のタイミングは、マウスにとっての朝、昼、夕、夜中とした。また投与後、0.5,  1, 1.5, および3時間後に血中の脂肪量を測定した。その結果、中鎖脂肪酸の中性脂肪(TG)は投与後いずれの時間でも血中濃度は低いままであった。また、投与タイミングでも、中鎖脂肪酸のTGはいずれの時刻でも血中濃度が大きく変動することはなかった。一方で、ラードの投与群は、1時間後をピークとするカーブが得られた。時間軸に対する中濃度下面積(AUC)を求めると、投与タイミングで大きく異なっていた。夜中から朝にかけてのラードの投与はAUCを大きくはしなかった。一方、昼、夕方投与は、AUCの値が大きくなることがわかった。この違いは、体内時計影響を強く受けていることに起因すると考えられる。夜中や朝は、マウスの摂食活動は低いところであるので、摂食した脂肪はエネルギーとして優先的に使われ、そのために血中のTGは低かったものと考えられる。一方、昼や夕方はマウスの摂食行動は盛んな時期なので、追加で投与した脂肪は使われないために血中に多く残った可能性が考えられる。また、魚油のAUC変化はラードと類似していることがわかった。以上のことをヒトの食生活あてはめて考察すると、朝食時の脂質は代謝される可能性があり、肥満になりにくいが、夕食時の脂質は代謝されにくく、脂肪蓄積回され、肥満を助長する可能性が示唆された。次にDHAとEPAを多く含む魚油の体内時計促進効果について調べた。その結果、大豆油に比較して魚油は体内時計のリセット効果が強いことがわかった。また、この作用にはGLP-1の分泌促進が関わっている可能性が示唆された。

  • 週間リズムの視点での運動科学・栄養科学

    2013  

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    食事・栄養と体内時計の関係を調べる学問を時間栄養学と称している。ところで、ヒトは1週間を単位として生活している。つまり、平日と週末という考え方、5日と2日の組み合わせによる行動とも捉えられる。このようリズムを週間リズムと呼ぶことにする。最近研究では平日と週末で睡眠時間の差が大きいほど、つまり、週末に寝だめする傾向の人は肥満症状を呈しやすいこと、また大学生を対象とした研究では成績が振るわないことがわかってきた。このように週間リズムの不調は社会的な時差ボケ状態になることが予想される。今回、マウスでこのようなモデルを作成した。高脂肪食を与える実験を行った。5日高脂肪食+2日普通食、2日高脂肪食+5日普通食、7日高脂肪食、7日普通食とした。その結果、5日高脂肪+2日普通食群では摂食パターンや体重変化に顕著な影響が見られた。5日の高脂肪食時には体重が増加し、次の2日の普通食日には摂食低下と体重減少が認められ、次の月曜日にリバンドの摂食増大が見られた。したがって、体重変化は7日高脂肪食群と差が見られなかった。次に自発的な輪回し運動において、7日運動なし、5日運動+2日なし、2日運動+5日なし、および7日なしの4群を作製した。高脂肪食による肥満に対して運動の抑制効果は弱かった。次に、先に認められた、5日高脂肪食+2日普通食のグループで、5日の高脂肪食時にのみ、輪回し運動を組み合わせる実験を行った。すなわち、運動の高肥満効果が期待できることと、運動が摂食抑制を引き起こす可能性が考えられた。高脂肪食と運動の組み合わせは、リバンド性の体重増加を顕著に抑制したが、摂食そのものリバンドは抑制できなかった。以上、食事性の週間リズム形成はマウスの系でも可能であること、また、週間リズムにはリバンド性の影響が強く出ることがわかった。運動はリバンド性の食欲は抑制できなかったが、体重増加は抑制できたので、ヒトの食生活改善に部分的役立つ知見であると考えられた。

  • 食物アレルギー発症・寛容におけるマウス体内時計の役割解明

    2012  

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    [背景・目的]日本をはじめとする先進国では,食物アレルギーを発症する人が急増しており,食物アレルギーは食品安全上重要な課題となっている。アレルギーと体内時計には深い関わりがあると示唆されており,例えば喘息では,夜から早朝にかけて副交感神経の亢進、交感神経の低下により,症状が悪化しやすいという時間生物学的特徴がある。しかし,食物アレルギーと時間との関連についてはあまり分かっていない。そこでマウスを用いて,食物抗原摂取時間の違いによりアレルギー症状に差が出るのかを検証した。また,症状の程度が時間により異なる原因について,体内時計が腸管防御機構にどのような影響を与えているのかを調べた。[方法] マウスの腹腔内にOVAを暴露し, ELISAによりOVA特異的IgEが産生され,感作されていることを確認した。6時間の絶食を経て,早朝に相当するZT9もしくは夕方に相当するZT21に,OVA80mgを含む水溶液を経口投与し,30-60分後に症状を観察した。症状は便と,経口投与前後での体重変化で評価した。3日後に2度目の投与をし,症状を観察した。観察後,腸間膜リンパ節を採取し培養を行った。細胞数を揃えOVAで再刺激をし,72時間後培養液を回収した。その後,上清中のIL-5およびIL-13をELISAにより測定した。もう一つの実験として,抗原の血中移入量が時間により異なるのかを調べた。抗原は腸から血液,リンパ管を経てリンパ器官へ移入し,免疫反応を起こす。そのため,血中の抗原量を測定するのは免疫反応の起こりやすさを推測するのに有効だと考えた。感作をしていないマウスに,先の実験同様6時間の絶食を経て,ZT9かZT21にOVA80mgを含む水溶液を経口投与し,30分後採血した。血清中のOVA量はELISAにより測定した。[結果]便は状態に応じてスコア化した。異常がない・便をしない場合は0,軟便である場合は1,下痢便である場合は2,下痢便で排便量も多くお尻が汚れている場合は3でスコア化した。ZT21群に比べてZT9群で便スコアが有意に高くなった。排便量が多い程体重は減少するはずであるが,便の状態と相関して体重が減少していることが確認できた。体重変化においてもZT9群で有意に体重が減少する結果となり,症状はZT9で悪化することが分かった。培養液中のIL-5とIL-13は,どちらもZT9群で多いという結果が得られた。抗原の血中移入量に関しても,ZT9群で有意に多いという結果が得られた。[考察]ZT9で症状が悪化した原因として,腸間膜リンパ節の培養実験から,IL-5やIL-13を分泌するTh2細胞がZT9で活性化するため,ということが示唆された。便スコアとの関係を見ると,分泌量は便スコアに依存していることが分かる。このことから,Th2細胞の働き具合は症状の程度を左右する要因の1つであると考えた。 また,抗原血中移入量がZT9で多いため,免疫反応が起こりやすくなり,これも症状が悪化する原因の1つだと考えられる。

  • アレルギー発症モデルマウスにおける体内時計の役割解明

    2010  

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    アレルギー疾患に体内時計がどのように関わっているかを明らかにする研究を行った。アレルギー症状としての花粉症や喘息は早朝から午前中にかけて症状が悪化する事が知られている。また、アレルギー性の痒みは夜間の寝る前に起こりやすいことも知られている。そこで、マウスを用いて、種々のアレルギー反応に体内時計がどのように関わっているかを、ヒスタミンによる皮膚透過性反応を指標にアレルギー反応について調べた。皮膚透過性反応にはリズムが見られ、夜間の初めは反応が小さく、夜間の後半から昼間にかけては強いことが分かった。次に時計遺伝子であるPer2遺伝子の変異マウスを用いて同様の実験を行ったところ、いずれの時間帯でも、透過性反応が強かった。血中のコーチコステロンのリズムならびに含量を測定したところ、ワイルドマウスでは、夜間の始めに血中のコーチコステロンの濃度が増大する一方、変異マウスはそのような増大が見られず、1日中同じ値であった。一方、IgE受容体感受性や発現量を調べた結果、両者に差は見られなかった。また、ヒスタミン分泌細胞である肥満細胞を培養し、両者のリズム性のヒスタミン分泌能を調べたところ、両者に差は見られなかった。以上の結果から、痒みのリズムを引き起こす原因は副腎皮質ホルモン分泌リズムに依存し、体内時計の異常は、コーチコステロンリズムの医異常を引き起こし、痒みを増大させる可能性が指摘された。 卵白アルブミン(OVA)の感作により、鼻炎モデルや喘息モデルが作られている。そこで、感作時間が異なることで、アレルギー反応が増大するか否かについて調べた。1日を4等分して、朝、昼、夕、夜にOVAを注射し、血中のIgE量を測定した。その結果、夕方感作が一番IgEの増大が小さいことが分かった。先に述べたようにコーチコステロンの可能性を検証するために、副腎摘出やデキサメサゾン投与下のIgEを測定すると、仮説通り、感作にも副腎皮質を介したリズムがあることが分かった。

  • マウス皮膚の時計遺伝子発現に対する環境刺激応答

    2008  

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    時計遺伝子の発現パターンの解析により、時計遺伝子は、中心時計である視交叉上核のみならず脳や末梢臓器に発現することがわかった。すなわち体内時計はシステムとして成り立っており、それぞれの臓器に発現している時計遺伝子が、ローカル時計として機能している可能性が指摘された。今回、マウスの皮膚に発現する時計遺伝子の機能を探る目的で、皮膚に対する温度刺激の応答性について、サーカディアンリズムの観点から調べた。(1)まず、皮膚に発現するPer1とPer2遺伝子発現リズムを調べた。その結果、顕著なリズムを示し、昼間低く、夜間高いという、これまでに知られていた肝臓のリズムパターンと類似したパターンを示すことが判明した。(2)次にマウスを37℃と41℃に30分間温浴させ、その後皮膚の時計遺伝子発現を調べた。Per1遺伝子発現は温度依存的に低下したが、Per2遺伝子発現には変化はみられなかった。したがって、Per1遺伝子発現の一過性の低下が同調刺激になる可能性が指摘された。そこで次に毎日の一定時刻の温度刺激が体内時計を同調させるか否かについて調べた。(3)Per2::luciferaseノックインマウスあるいはBmal1::luciferaseのトランスゼニックマウスを用いて、皮膚や肝臓のluciferaseによる生物発光リズムの位相を測定することで、温度刺激が位相変容作用を示すか否かについて調べた。1日を4等分して、朝、昼、夕方、夜間のいずれかに、マウスを41℃で温浴を30分行わせ、この操作を5日間行った。その後、マウスの皮膚や肝臓を取り出し、ルミサイクルという装置を用いて、生物発光リズムを調べた。その結果、朝や夜間に温浴させたマウスの時計遺伝子発現リズム位相が前進していた。すなわち人に外挿すると、夕方から就寝にかけて風呂に入る習慣がついている人は、夜型から朝型に移せる可能性が示唆された。

  • マウス中枢神経時計遺伝子発現に対するドパミン制御機構

    2007  

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    体内時計に関わる時計遺伝子は脳に広く発現することが知られている。ドパミン神経の起始核である黒質や中脳被蓋野にも時計遺伝子が発現し、ドパミンの時間機能に関わっていることが知られている。ドパミン神経は報酬系に関わっているため、覚醒剤やコカインの依存性さらにアルコールやニコチンの依存性にも時計機構が関与する可能性がある。そこで今回、時計遺伝子が変異を起こしているPer2ミュータントとClockミュータントを用いて、覚醒剤のメタンフェタミンに対する作用を調べた。その結果、Per2ミュータントのメタンフェタミンに対する行動量増加作用や、依存性に関わるsensitizationについてはワイルドマウスと同程度であり、Per2変異の影響は小さかった。つぎに同様の実験をClock変異マウスを用いて行った。その結果Per2変異マウスと同様にメタンフェタミンの作用にClock変異の影響は見られなかった。次にドパミン神経の活性に関わるドパミントランスポーターの遺伝子発現について調べた。その結果、トランスポーターの遺伝子発現には顕著なリズムが認められ、Per2変異マウスではリズム性が減弱していた。そこでメタンフェタミンのドパミントランスポーターの作用発現に対する作用を日内リズムの観点から調べた。このトランスポーターの発現が低い昼間にメタンフェタミンを投与すると顕著な遺伝子発現の増大が認められ、逆にトランスポーターの基礎発現量が高い時刻にメタンフェタミンを投与しても殆ど影響されなかった。以上の結果、メタンフェタミンの作用には投与時刻による作用の違いが出る可能性が強く示唆された。そこで、Per2遺伝子欠損マウスとワイルドマウスに昼間と夜間にメタンフェタミンを投与すると、ワイルドマウスでは明らかに昼間の投与が顕著であったが、夜間の活動期では作用が弱く、このような昼夜差は欠損マウスでは認められなかった。以上、体内時計はドパミン神経に作用する薬物の昼夜差に積極的に関与することが明らかとなった。

  • マウス体内時計遺伝子発現リズムの組織・器官特異的同調刺激の解明

    2006  

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    体内時計遺伝子は、中心時計の視交差上核のみに発現するのみならず、大脳皮質などの脳部位さらに、末梢臓器の肺や肝臓にも発現することが知られている。ところで視交差上核の主時計、大脳皮質の脳時計、末梢臓器の末梢時計が、同一の情報シグナルで同調されるのか、あるいは部位特異的な同調機構で同調されるのかを明らかにすることは、リズム不調の疾病やその薬物開発に重要な意味となる。主時計の視交差上核は外界の光刺激で同調されることは良く知られている。しかしながら、光照射を行っても直ちに大脳皮質や肝臓などの体内時計発振は同調されないことがわかった。すなわち、主時計以外の時計では、どのような生体シグナルが同調として働くか不明なままである。本研究から大脳皮質はメタンフェタミンやニコチンなどの覚醒刺激を起こす薬剤で同調されやすいことがわかった。実際、毎日一定時刻にメタンフェタミンを腹腔内投与したり、ニコチンを噴霧して与えた場合、大脳皮質や線条体の時計遺伝子発現リズムは同調され、腹腔内投与や噴霧投与の時刻依存的なピーク時刻が出現した。一方、末梢臓器に発現するリズムは大きく2つの因子で同調されることがわかった。すなわち、食事のシグナルとステロイドホルモンのシグナルでの同調である。食事時刻を変えると、視交差上核を破壊していても、肝臓や肺ではその時刻依存的リズムのピーク位相が出現した。また、肝臓や肺では、副腎皮質ステロイドホルモンを毎日1回噴霧投与すると、これでも同調されることがわかった。一方、大脳皮質ではステロイドホルモンでは同調されなかった。以上の実験成績をまとめると、大脳皮質の時計は覚醒刺激が同調因子となり、末梢臓器では食事やそれにともない変化する副腎皮質ステロイドホルモンが同調因子となることが判明した。

  • マウス時計遺伝子群のドパミン神経制御機構

    2006  

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    本研究は2つの視点から成り立っている。ひとつは体内時計がドパミン神経の発現機構に影響を及ぼすかという視点である。もうひとつはドパミン神経の活性化が体内時計の同調因子となりうるかという視点である。まず前者の例として、ドパミン活性の指標としてドパミントランスポーター(DT)の発現リズムを調べた。その結果、ドパミン神経の豊富な線条体領域で、顕著なDTのmRNAリズムが認められた。特に夜間の活動期に高値を示した。体内時計のミュータント動物であるClock-/-, mPer2-/-のマウスでは、このようなリズムが消失した。以上の結果は、体内時計がドパミンの神経活性にリズム性の影響を及ぼすことを示している。次にドパミン神経の活性化や不活性化が体内時計に及ぼす影響について調べた。メタンフェタミンを飲水投与して出現するメタンフェタミン性リズムは線条体の6-OHDAによるドパミン破壊や熱凝固破壊により消失した。このことはこのリズム形成はドパミン神経が重要な役割を演じていることを意味する。次に線条体のドパミン神経細胞の起始核である黒質の破壊によってもメタンフェタミン性のリズムは消失した。このことは黒質―線条体のドパミン神経が正常でないと、メタンフェタミン性のリズム形成が起こらないことを意味した。次にオレキシン神経が破壊され、覚醒レベルが低下したマウスを用いてメタンフェタミン性のリズム形成を調べると、リズム形成が低下していた。このマウスの線条体のDTの発現を調べると低下し、メタンフェタミン投与によっても影響されなくなってしまった。以上の結果をまとめると、体内時計とドパミン神経は非常に密接な関係があり、メタンフェタミン性リズムがヒトの睡眠覚醒リズムを反映していることを考えると、黒質―線条体ドパミン神経系の睡眠リズムの形成に重要な脳部位といえる。

  • 時刻認知の学習・記憶形成における時計遺伝子の働き解明

    2005  

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    本研究では時遺伝子が予知行動形成にどのようにかかわるかを明らかにすることを研究の第一目標に掲げた。 Cry1/Cry2(無周期)、mPer2(短周期)、Clock(長周期)の時計遺伝子ミュータントマウスが、それぞれ、給餌サイクルの周期(22-28時間)を変えた刺激に反応して予知行動を形成するか否か調べこととした。面白いことに、それぞれのミュータントのフリーラン周期に応じて、mPer2ミュータントでは短周期の給餌リズムに同調し、Clockミュータントでは長周期に同調する傾向が見られた。そこで、Clockミュータントで26時間周期に対して予知行動形成が完成した動物を用いて、この動物の脳を6時間間隔で摘出し、mPer2やClock以外のBmal1, mPer1,などの遺伝子発現変動リズムを調べた。さらに、体内時計の下流遺伝子である、Dbp ,E4bp4などの遺伝子発現についても調べた。その結果、Clockミュータントで予知行動に同調したマウスでは、肝臓や、大脳皮質の時計遺伝子の位相が、26時間周期に合っていた。一方、Wildマウスでは24時間周期の給餌提示に合いやすく、その結果、肝臓や大脳皮質の時計遺伝子発現リズムも24時間周期を示した。一方、Cry1/Cry2のミュータントマウスでも、予知行動が出現するようであれば、予知行動リズム形成にかかわる時計遺伝子は先に述べた従来の時計遺伝子とは異なった遺伝子発現を伴う可能性がある。実験結果では、Cry1/Cry2のミュータントマウスでは予知行動が減弱され、また不安定になる傾向にあったが、阻害されなかった。以上の結果、従来の時計遺伝子も予知行動リズム形成にかかわっていることが明らかになったが、その関与は弱いものと思われる。なぜなら、ほとんど無周期を示すCry1/Cry2のミュータントマウスでもリズム形成は可能であったからである。

  • 内外環境変化に対するマウス体内時計の適応応答

    2005  

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    不登校や自閉症といった疾病に体内時計異常に基づく睡眠障害が出ることがよく知られている。時計遺伝子ミュータントマウスの母親と子供を用いて、内外環境変動が体内時計のリズム形成にどのように働くかについて調べた。本研究では特にリズムの発達障害に対する内的環境変動として、(1)母親の体内時計異常が子供の体内時計発達にどのような影響を及ぼすか点と、外的環境変動として、(2)子供の飼育環境の光環境を恒常明や恒常暗にすることが子供の体内時計の発達にどのような影響を及ぼすかを調べた。WildとClockミュータントの母親を用い、仔マウスの入れ替え実験を行い、その後、成長した後これらのマウスの行動リズム障害を調べた。その結果、母親の体内時計に異常があると、母性行動や授乳行動リズムに異常が生じ、その結果、仔マウスの発達やリズム形成に障害が出てくることが明らかとなった。また、母性行動形成にかかわるホルモンとして知られている、プロラクチンの分泌リズムを調べたところ、Clockの母親は分泌量の低下とリズムの平坦化が認められた。このように母親のリズムが不調だと子育てに影響することが明白になった。つぎに、仔マウスを恒常明や恒常暗で飼育した。その結果、Wildマウスではいずれの飼育条件下でも、行動リズム形成にはなんらの影響も受けなかった、Clockミュータントの仔マウスは、離乳までの時期に恒常明飼育を受けると、行動開始位相に遅れが出ることが判明した。つまり、Clock遺伝子の異常に外的要因として発達期の光環境が影響すると、睡眠相後退症候群(DSPS)のマウスモデルができることがわかった。このようなマウスに臨床的にも使用されているメラトニンを投与するするとDSPS状態が改善された。

  • 情動制御におけるオレキシン神経の役割解明

    2005  

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    オレキシン神経は覚醒・睡眠や摂食行動にかかわっていることが広く知られている。先の研究で、オレキシン神経欠損マウスは、給餌制限に伴う予知行動リズム形成が減弱することが報告されている。予知行動リズムは視交差上核を破壊しても影響されないことから、視交差上核の機能を伴わないリズムとして知られている。このようなリズムの他の一つとして、メタンフェタミン(MA)性リズムが知られている。MAを飲水中に添加し与えると、マウスやラットは明暗環境下にもかかわらず、24時間周期より長いリズム示すことが知られている。先に述べたようにオレキシン神経は覚醒に深く関与する分子であるので、MAのリズム形成に影響を及ぼす可能性が考えられる。オレキシン神経欠損マウスにMAを0.005%で飲水投与を行なったところ、対照マウスに比較して、MAリズム形成が顕著に障害された。また、オレキシン欠損マウスは、MAの急性投与による行動量の上昇も低下していた。さらにMAを飲水投与したときの行動量の増大も減弱していた。ところで、オレキシン神経が、DA神経の起始核である中脳被蓋野に促進性に投射していることがわかっているので、オレキシン神経欠損マウスのMAリズム形成障害はDA神経の機能低下に基づく可能性が指摘できる。そこで、中脳被蓋野のDA神経を6-OHDAで破壊し、MAを飲水投与させた。その結果、破壊マウスは、対照群に比較して、リズム形成が非常に悪かった。MAによるリズム形成を確認した後、マウスの線条体や側座核のTH(DA産生の律速酵素)の発現量を免疫組織化学で調べた。6-OHDA破壊マウスの側座核のTHの発現量は著明に低下していた。以上の結果をまとめると、オレキシン神経は中脳被蓋野―側座核のDA神経の活性化を通してMA性リズム形成に関与していることが明らかとなった。

  • 時刻認知の予知行動形成における時計遺伝子の働き解明

    2004  

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    マウスやラットに毎日一定時刻の給餌を行うと、動物は餌を得られる時刻に先立って、2-3時間前より活動量が高くなる。このような行動は毎日決まった時間に出現することから、予知行動リズムという。予知行動リズムは給餌時間を24時間よりずれた周期で行うと形成できないことから、このリズム形成には体内時計がかかわっていることが想定される。ところで、最近の研究から数多くの体内時計遺伝子が見出されてきているが、特にCry1とCry2遺伝子の同時ノックアウトは、恒暗条件下に飼育するとリズム性を失うことが知られている。このようにリズム性を失った状態でも予知行動リズムが形成されるか否かについては興味が持たれる。なぜなら、給餌性リズム形成に従来から知られている時計遺伝子が関与するのか、あるいは違った分子がかかわっているのかを明らかにすることができるからである。Cry1/Cry2のダブルノックアウト動物を、正常明暗状態出飼育し、もしくは、恒暗条件にし、視交叉上核に基づくリズム性をなくした状態で、あるいは、視交叉上核を電気破壊し、視交叉上核性のリズムを完全に止めた状態で、給餌制限した。給餌制限による餌の獲得量には、Wild群と差は見られなかった。ところが、上の3条件のいずれの実験においても、強弱は存在するものの、給餌制限による予知行動リズムが形成できた。したがって、このリズム形成には本質的にはCry分子を中心とする負の制御機構は必要ないことがわかった。つまり、給餌性の予知行動リズム形成には視交叉上核を中心とした体内時計システムとは異なったシステムが存在する可能性が指摘できた。しかしながら、Wildに比較すると形成が遅くかつ、不安定なものであった。したがって、正常な予知行動リズム形成には、Cryの時計遺伝子も少なからず、寄与していると結論した。

  • 時計遺伝子利用による分子時間薬理学研究の新展開

    2001  

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    臨床的に心筋梗塞の発症時間が午前中に多いことが知られているが、その詳細なメカニズムについては分かっていない。ところで、血液凝固―線溶系にプラスミンが深く関わっているが、プラスミノーゲンアクチベータインヒビター1(PAI-1)の活性化はプラスミンの産生を低下させ血液の粘性を高めると考えられる。PAI-1の遺伝子発現には明瞭な日内リズムがあり、人の場合明け方に、夜行性動物の場合夕方に発現量が増大する。このような研究から、PAI-1遺伝子発現が体内時計遺伝子の制御下にある可能性が強く示唆される。(1)正常マウスならびに時計遺伝子clockの欠損マウスを用いて、心臓におけるPAI-1発現のサーカディアンリズムについて調べた。(2)制限給餌時のPAI-1遺伝子発現リズムの変化を調べた。さらに、(3)時間薬理学的見地から、アドレナリン投与や不動、絶食ストレスさらにトレッドミル、強制水泳などをPAI-1遺伝子発現が高い時刻と低い時刻に処置し、PAI-1発現にどのような影響を及ぼすかについて調べた。(1)PAI-1遺伝子発現は夜間の始めをピークとする日内リズムが見られ、clockのミュータント動物ではそのリズムの振幅が消失していた。(2)給餌時刻を昼間に設定することにより心臓のmPer遺伝子発現のピークも昼間に移行したが、それと同時にPAI-1遺伝子発現も昼間にピークが来た。(3)アドレナリンの投与はいずれの時間帯もとくに発現が低い時間帯でPAI-1遺伝子発現を増大させたが、既に発現が高い時刻でも、それ以上に発現を増大させた。また、不動ストレス、絶食ストレスでも同様な結果を得たが、運動刺激はなんら影響を及ぼさなかった。PAI-1遺伝子発現は一部体内時計の支配下にある可能性が示唆された。また臨床的に、心筋梗塞のリスクファクターが高い時刻においては交感神経の活性化やストレスに対して注意する必要性が強く示唆された。

  • 動物サーカディアンリズムに対するストレス負荷の影響

    1997  

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    ストレスが蔓延する現代社会では、心身症、うつ病、神経症などストレス性疾患が増大している。これらの疾患には睡眠障害を始めとするリズム性の異常も多い。したがって、本研究はこのようなリズム異常に関わる疾患の発症の原因を探る目的の一つとして、ストレスの光同調に対する影響について調べた。ハムスターを恒暗条件飼育し、フリーランリズムが出現してきたらサーカディアンタイム(CT)(CT12を輪回し行動の開始時刻とする)の13や20に光照射を行った。光照射のみをCT13やCT20行ったハムスターの位相後退や前進はそれぞれ74分と149分であった。拘束ハムスターに光照射すると位相後退も前進もいづれもその大きさが有意に低下した。 以前フットショックストレスを経験したハムスターは対照群に比較してCT13やCT20の光照射によるそれぞれ位相後退や前進が少なかった。フットショックストレスのみを負荷した群(stress)とフットショックストレスも光照射も与えなかった群(control)の視交差上核内FOS免疫陽性細胞数はそれぞれ平均19と21で有り、両群間に全く差は認められなかった。すなわち、フットショックストレスは視交差上核の細胞にFOSを出現させないことがわかった。一方、光照射した群では光照射しない群より、FOS免疫陽性細胞数(45)が有意に増大した。この光照射によるFOS免疫陽性細胞数の増加はフットショックストレスの併用により有意に低下し(19.7)、無処置群やストレスのみの処置群の値に近づいた。 本研究において、物理的な拘束ストレスでも、また無条件恐怖刺激や条件恐怖刺激といったフットショックストレスでも、光照射によるハムスターの輪回しリズムの位相変化を抑制することが明らかとなった。また、フットショックストレスは光刺激による視交差上核内FOS免疫陽性細胞数の増大を抑制した。光同調時にストレスによるセロトニン神経の活性化が起こり、結果として光同調を低下させたものと考えられる。発表論文:柴田重信、光とメラトニン、ファルマシア34、45、1998池田真行、柴田重信、概日リズムとセロトニン神経系、Molecular Medicine,34,286,1997

  • 老化による体内時計への光同調低下の神経機構解明

    1996  

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     外界の光刺激は動物の体内時計をリセットするが、光同調が老化により減弱している可能性がある。視交叉上核シナプスの長期増強現象をはじめとする可塑的変化に老化の影響があるか否かについて研究することは、外界の光刺激による同調の神経機構そのものを理解する上にも、また老化による同調性の低下のメカニズムを理解するにも重要なことだと考えられる。  光刺激の情報は視交叉上核のVIP ニューロンに伝達されると考えられている。そこで視交叉上核からのVIP遊離を指標として、視神経刺激がこの遊離を増大させるか否かについて調べた。また、VIP遊離に対する老化の影響について調べ、視交叉上核シナプス伝達の老化による低下をVIP遊離という物質的背景についても調べた。視神経刺激により視交叉上核からのVIP遊離は増大した。VIP遊離の増大は視神経刺激の強度と周波数に比例して増大した。すなわち、0.9mA、20Hzの電気刺激によりVIPの遊離量は基礎遊離量に対して約130-150%に増大した。老齢動物の視交叉上核からのVIPの基礎遊離は若齢群とほぼ同程度であった。しかしながら、視神経刺激や高濃度KClによるVIPの遊離増強は減弱した。視交叉上核のシナプス機構を反映すると考えられる長期増強現象に対する老化の影響を調べた。視神経刺激により視交叉上核から記録される電場電位の大きさは時刻依存性に高頻度刺激(100Hz、1秒間)により増大した。明期に高頻度刺激を行うと電場電位の大きさは刺激後60-120分後には刺激前の約200%に増大した。またこの増大現象は刺激直後には現れず、ゆるやかなカーブを示した。以上の結果、シナプス伝達機構が老化により低下することが明らかとなりこの低下は老齢動物の光同調性の低下に関連するものと考えられた。このようなシナプス伝達の低下を改善する薬物が見つかれば、これは老化による光同調の低下を軽減させうるものと考えられる。

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  • 体内時計の研究 時間栄養学と時間運動学の研究の研究動向調査

    2015.06
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    2015.08

    フランス   ニース大学

 

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