Updated on 2026/09/21

写真a

 
KUBO, Takeo
 
Affiliation
Faculty of Science and Engineering, Graduate School of Advanced Science and Engineering
Job title
Professor(non-tenure-track)
Degree
博士（薬学）

Research Experience

  • 2026.04
    -
    Now

    Waseda University   Department of Integrative Bioscience and Biomedical Engineering, Graduate School of Advanced Scienece and Engineering   Professor (Fixed-term)

  • 2001.04
    -
    2026.03

    The University of Tokyo   Graduate School of Science, Department of Biological Sciences

  • 1997.04
    -
    2001.03

    The University of Tokyo   Graduate School of Pharmaceutical Sciences

  • 1995.07
    -
    1997.03

    The University of Tokyo   Faculty of Pharmaceutical Sciences

  • 1992.12
    -
    1995.06

    The University of Tokyo   Faculty of Pharmaceutical Sciences

  • 1987.11
    -
    1992.11

    The University of Tokyo   Faculty of Pharmaceutical Sciences

▼display all

Research Areas

  • Molecular biology / Developmental biology / Neuroscience-general / Animal physiological chemistry, physiology and behavioral biology

Research Interests

  • ケニヨン細胞

  • キノコ体

  • 尻振りダンス

  • 社会性行動

  • ミツバチ

  • アフリカツメガエル

  • 器官再生

▼display all

Awards

  • 日本動物学会賞

    2024   公益社団法人 日本動物学会  

  • Zoological Award （論文賞）

    2017   公益社団法人 日本動物学会  

    Winner： 河野大輝氏他との共同受賞

  • JB論文賞

    1994   公益社団法人 日本生化学会  

    Winner： 南部りか氏他との共同受賞

 

Papers

  • Putative muscle stem cells promote Xenopus tail regeneration by modifying macrophage function via c1qtnf3

    Sumika Kato, Takeo Kubo, Taro Fukazawa

    Proceedings of the National Academy of Sciences   122 ( 47 )  2025.11

    　View Summary

    In Xenopus laevis tadpole tail regeneration, lineage-restricted tissue stem cells produce differentiated cells that form regenerated tail tissues, but the behavioral dynamics of tissue stem cells during tail regeneration remain largely unknown. We previously reported that multiple tissue stem/progenitor cells can be efficiently enriched from regeneration buds using the side population (SP) method. Here, we performed trajectory inference using single-cell RNA sequencing data of the SP fraction to construct differentiation trajectories and identify putative tissue stem cell populations that initiate differentiation pathways. We found that complement c1q tumor necrosis factor-related protein 3 ( c1qtnf3 ) is specifically expressed in putative muscle stem cells (MSC) and, using knockdown (KD; CRISPR/Cas9-based F0 crispants) experiments, demonstrated that c1qtnf3 is necessary for tail regeneration. Furthermore, we found that the impaired tail regeneration by c1qtnf3 KD was accompanied by abrogation of macrophage-like cell accumulation at the amputation site. These phenotypes were rescued by macrophage-like cell-specific forced expression of neutrophil cytosolic factor 1 , a gene related to effector molecule production in myeloid cells, suggesting that the impaired tail regeneration by c1qtnf3 KD is due to macrophage dysregulation. Our findings suggest that, in Xenopus , putative MSC modulate macrophage function via c1qtnf3 expression for successful tail regeneration.

    DOI

  • Analysis of molecular and cellular bases of honey bee mushroom body development.

    Shuichi Kamata, Takeo Kubo, Hiroki Kohno

    Scientific reports   15 ( 1 ) 21462 - 21462  2025.07  [International journal]

    　View Summary

    In the honey bee, mushroom bodies (MBs), a higher-order center of the insect brain, comprise three class I Kenyon cell (KC) subtypes (lKC, mKC, and sKC) with distinct somata sizes and locations and gene expression profiles. While these KC subtypes have been suggested to function in different behavioral regulations, the molecular and cellular basis of their development remains obscure. Here, we showed that lKCs, mKCs, and sKCs are produced in that order at different pupal stages by labeling proliferating MB cells with 5-ethynil-2'-deoxyuridine at various pupal stages. RNA-sequencing analysis of FACS-sorted pupal MB cells identified genes that were upregulated in proliferating and non-proliferating MB cells, respectively. Furthermore, in situ hybridization of some of these genes labeled the proliferating cells or immature KCs in the MBs at pupal stages producing each subtype. We found that the expression patterns of SoxNeuro, optix, and asense were consistent with those in Drosophila MBs, while odd-paired, which functions in neuroblasts in Drosophila, was preferentially expressed in immature KCs in honey bees. Our findings revealed the basic scheme of the molecular and cellular processes of honey bee MB development and suggested that they are at least partially different from those of Drosophila MB development.

    DOI PubMed

  • Unique spatially and temporary-regulated/sex-specific expression of a long ncRNA, Nb-1, suggesting its pleiotropic functions associated with honey bee lifecycle.

    Hiroto Tadano, Hiroki Kohno, Hideaki Takeuchi, Takeo Kubo

    Scientific reports   14 ( 1 ) 8701 - 8701  2024.04  [International journal]

    　View Summary

    Honey bees are social insects, and each colony member has unique morphological and physiological traits associated with their social tasks. Previously, we identified a long non-coding RNA from honey bees, termed Nb-1, whose expression in the brain decreases associated with the age-polyethism of workers and is detected in some neurosecretory cells and octopaminergic neurons, suggesting its role in the regulation of worker labor transition. Herein, we investigated its spatially and temporary-regulated/sex-specific expression. Nb-1 was expressed as an abundant maternal RNA during oogenesis and embryogenesis in both sexes. In addition, Nb-1 was expressed preferentially in the proliferating neuroblasts of the mushroom bodies (a higher-order center of the insect brain) in the pupal brains, suggesting its role in embryogenesis and mushroom body development. On the contrary, Nb-1 was expressed in a drone-specific manner in the pupal and adult retina, suggesting its role in the drone visual development and/or sense. Subcellular localization of Nb-1 in the brain during development differed depending on the cell type. Considering that Nb-1 is conserved only in Apidae, our findings suggest that Nb-1 potentially has pleiotropic functions in the expression of multiple developmental, behavioral, and physiological traits, which are closely associated with the honey bee lifecycle.

    DOI PubMed

  • Analysis of Antennal Responses to Motion Stimuli in the Honey Bee by Automated Tracking Using DeepLabCut

    Hiroki Kohno, Shuichi Kamata, Takeo Kubo

    Journal of Insect Behavior    2024.01

    　View Summary

    Abstract

    Considering recent developments in gene manipulation methods for honey bees, establishing simple and robust assay systems which can analyze behavioral components in detail inside a laboratory is important for the rise of behavioral genetics in the honey bee. We focused on the antennal movements of the honey bee and developed an experimental system for analyzing the antennal responses (ARs) of the honey bee using DeepLabCut, a markerless posture-tracking tool using deep learning. The tracking of antennal movements using DeepLabCut during the presentation of vertical (downward and upward) motion stimuli successfully detected the direction-specific ARs in the transverse plane, which has been reported in the previous studies where bees tilted their antennae in the direction opposite to the motion stimuli. In addition, we found that honey bees also exhibited direction-specific ARs in the coronal plane in response to horizontal (forward and backward) motion stimuli. Furthermore, an investigation of the developmental maturation of honey bee ARs showed that ARs to motion stimuli were not detected in bees immediately after emergence but became detectable through post-emergence development in an experience-independent manner. Finally, unsupervised clustering analysis using multidimensional data created by processing tracking data using DeepLabCut classified antennal movements into different clusters, suggesting that data-driven behavioral classification can apply to AR paradigms. In summary, our results revealed direction-specific ARs even in the coronal plane to horizontal motion stimuli and developmental maturation of ARs for the first time, and suggest the efficacy of data-driven analysis for behavioral classification in behavioral studies of the honey bee.

    DOI

  • Possible functions of ecdysone signaling reiteratively used in the adult honey bee brain

    Yasuhiro Matsumura, Hiroki Kohno, Takeo Kubo

    Frontiers in Bee Science   1  2023.09  [Refereed]  [Invited]

    　View Summary

    The European honey bee is a model organism for investigating the molecular and neural bases of the brain underlying social behaviors. Mushroom bodies (MBs) are a higher-order center of memory, learning, and sensory integration in insect brains, and honey bee MBs are a model to study adult neuronal plasticity. In the honey bee, MBs comprise three Class I Kenyon cell (KC) subtypes: large-, middle-, and small-type KCs, which are distinguished based on the size and localization of their somata, and gene expression profiles. One of the unique characteristics of honey bee MBs is that genes for ecdysone signaling are expressed in a spatially and temporarily regulated manner in the adult brain, suggesting that they play a role in the functional specialization of each KC subtype and behavioral control. A recent study reported that the transcription factor Mblk-1/E93, which functions downstream of ecdysone signaling during metamorphosis, targets genes involved in synaptic plasticity underlying memory and learning ability in the adult honey bee brain. On the other hand, the ecdysone receptor (EcR), which is expressed in small-type KCs in the MBs, was reported to target genes involved in lipid metabolism in the brain during foraging flight. The target genes for Mblk-1 and EcR in the adult brains differed from those during metamorphosis, implying that the reiterative use of some transcription factors involved in ecdysone signaling, such as EcR and Mblk-1, has contributed to the acquisition of novel MB functions in Aculeata species, including the honey bee.

    DOI

  • regeneration factors expressed on myeloid expression in macrophage-like cells is required for tail regeneration in Xenopus laevis tadpoles

    Momoko Deguchi, Taro Fukazawa, Takeo Kubo

    Development   150 ( 15 )  2023.07

    　View Summary

    ABSTRACT

    Xenopus laevis tadpoles can regenerate whole tails after amputation. We have previously reported that interleukin 11 (il11) is required for tail regeneration. In this study, we have screened for genes that support tail regeneration under Il11 signaling in a certain cell type and have identified the previously uncharacterized genes Xetrov90002578m.L and Xetrov90002579m.S [referred to hereafter as regeneration factors expressed on myeloid.L (rfem.L) and rfem.S]. Knockdown (KD) of rfem.L and rfem.S causes defects of tail regeneration, indicating that rfem.L and/or rfem.S are required for tail regeneration. Single-cell RNA sequencing (scRNA-seq) revealed that rfem.L and rfem.S are expressed in a subset of leukocytes with a macrophage-like gene expression profile. KD of colony-stimulating factor 1 (csf1), which is essential for macrophage differentiation and survival, reduced rfem.L and rfem.S expression levels and the number of rfem.L- and rfem.S-expressing cells in the regeneration bud. Furthermore, forced expression of rfem.L under control of the mpeg1 promoter, which drives rfem.L in macrophage-like cells, rescues rfem.L and rfem.S KD-induced tail regeneration defects. Our findings suggest that rfem.L or rfem.S expression in macrophage-like cells is required for tail regeneration.

    DOI

  • Identification of ecdysone receptor target genes in the worker honey bee brains during foraging behavior.

    Shiori Iino, Satoyo Oya, Tetsuji Kakutani, Hiroki Kohno, Takeo Kubo

    Scientific reports   13 ( 1 ) 10491 - 10491  2023.06  [International journal]

    　View Summary

    Ecdysone signaling plays central roles in morphogenesis and female ovarian development in holometabolous insects. In the European honey bee (Apis mellifera L.), however, ecdysone receptor (EcR) is expressed in the brains of adult workers, which have already undergone metamorphosis and are sterile with shrunken ovaries, during foraging behavior. Aiming at unveiling the significance of EcR signaling in the worker brain, we performed chromatin-immunoprecipitation sequencing of EcR to search for its target genes using the brains of nurse bees and foragers. The majority of the EcR targets were common between the nurse bee and forager brains and some of them were known ecdysone signaling-related genes. RNA-sequencing analysis revealed that some EcR target genes were upregulated in forager brains during foraging behavior and some were implicated in the repression of metabolic processes. Single-cell RNA-sequencing analysis revealed that EcR and its target genes were expressed mostly in neurons and partly in glial cells in the optic lobes of the forager brain. These findings suggest that in addition to its role during development, EcR transcriptionally represses metabolic processes during foraging behavior in the adult worker honey bee brain.

    DOI PubMed

  • Evolutionary dynamics of mushroom body Kenyon cell types in hymenopteran brains from multifunctional type to functionally specialized types.

    Takayoshi Kuwabara, Hiroki Kohno, Masatsugu Hatakeyama, Takeo Kubo

    Science advances   9 ( 18 ) eadd4201  2023.05  [International journal]

    　View Summary

    Evolutionary dynamics of diversification of brain neuronal cell types that have underlain behavioral evolution remain largely unknown. Here, we compared transcriptomes and functions of Kenyon cell (KC) types that compose the mushroom bodies between the honey bee and sawfly, a primitive hymenopteran insect whose KCs likely have the ancestral properties. Transcriptome analyses show that the sawfly KC type shares some of the gene expression profile with each honey bee KC type, although unique gene expression profiles have also been acquired in each honey bee KC type. In addition, functional analysis of two sawfly genes suggested that the functions in learning and memory of the ancestral KC type were heterogeneously inherited among the KC types in the honey bee. Our findings strongly suggest that the functional evolution of KCs in Hymenoptera involved two previously hypothesized processes for evolution of cell function: functional segregation and divergence.

    DOI PubMed

  • Mblk-1/E93, an ecdysone related-transcription factor, targets synaptic plasticity-related genes in the honey bee mushroom bodies.

    Yasuhiro Matsumura, Taiko Kim To, Takekazu Kunieda, Hiroki Kohno, Tetsuji Kakutani, Takeo Kubo

    Scientific reports   12 ( 1 ) 21367 - 21367  2022.12  [International journal]

    　View Summary

    Among hymenopteran insects, aculeate species such as bees, ants, and wasps have enlarged and morphologically elaborate mushroom bodies (MBs), a higher-order brain center in the insect, implying their relationship with the advanced behavioral traits of aculeate species. The molecular bases leading to the acquisition of complicated MB functions, however, remains unclear. We previously reported the constitutive and MB-preferential expression of an ecdysone-signaling related transcription factor, Mblk-1/E93, in the honey bee brain. Here, we searched for target genes of Mblk-1 in the worker honey bee MBs using chromatin immunoprecipitation sequence analyses and found that Mblk-1 targets several genes involved in synaptic plasticity, learning, and memory abilities. We also demonstrated that Mblk-1 expression is self-regulated via Mblk-1-binding sites, which are located upstream of Mblk-1. Furthermore, we showed that the number of the Mblk-1-binding motif located upstream of Mblk-1 homologs increased associated with evolution of hymenopteran insects. Our findings suggest that Mblk-1, which has been focused on as a developmental gene transiently induced by ecdysone, has acquired a novel expression pattern to play a role in synaptic plasticity in honey bee MBs, raising a possibility that molecular evolution of Mblk-1 may have partly contributed to the elaboration of MB function in insects.

    DOI PubMed

  • Cellular responses in the FGF10-mediated improvement of hindlimb regenerative capacity in Xenopus laevis revealed by single-cell transcriptomics.

    Nodoka Yanagi, Sumika Kato, Taro Fukazawa, Takeo Kubo

    Development, growth & differentiation   64 ( 6 ) 266 - 278  2022.08  [Domestic journal]

    　View Summary

    Xenopus laevis tadpoles possess regenerative capacity in their hindlimb buds at early developmental stages (stages ~52-54); they can regenerate complete hindlimbs with digits after limb bud amputation. However, they gradually lose their regenerative capacity as metamorphosis proceeds. Tadpoles in late developmental stages regenerate fewer digits (stage ~56), or only form cartilaginous spike without digits or joints (stage ~58 or later) after amputation. Previous studies have shown that administration of fibroblast growth factor 10 (FGF10) in late-stage (stage 56) tadpole hindlimb buds after amputation can improve their regenerative capacity, which means that the cells responding to FGF10 signaling play an important role in limb bud regeneration. In this study, we performed single-cell RNA sequencing (scRNA-seq) of hindlimb buds that were amputated and administered FGF10 by implanting FGF10-soaked beads at a late stage (stage 56), and explored cell clusters exhibiting a differential gene expression pattern compared with that in controls treated with phosphate-buffered saline. The scRNA-seq data showed expansion of fgf8-expressing cells in the cluster of the apical epidermal cap of FGF10-treated hindlimb buds, which was reported previously, indicating that the administration of FGF10 was successful. On analysis, in addition to the epidermal cluster, a subset of myeloid cells and a newly identified cluster of steap4-expressing cells showed remarkable differences in their gene expression profiles between the FGF10- or phosphate-buffered saline-treatment conditions, suggesting a possible role of these clusters in improving the regenerative capacity of hindlimbs via FGF10 administration.

    DOI PubMed

  • Effective enrichment of stem cells in regenerating Xenopus laevis tadpole tails using the side population method.

    Sumika Kato, Takeo Kubo, Taro Fukazawa

    Development, growth & differentiation   64 ( 6 ) 290 - 296  2022.08  [Domestic journal]

    　View Summary

    Xenopus laevis tadpoles have a strong regenerative ability and can regenerate their whole tails after tail amputation. Lineage-restricted tissue stem cells are thought to provide sources for the regenerating tissues by producing undifferentiated progenitor cells in response to tail amputation. However, elucidating the behavioral dynamics of tissue stem cells during tail regeneration is difficult because of their rarity, and there are few established methods of isolating these cells in amphibians. Here, to detect and analyze rare tissue stem cells, we attempted to enrich tissue stem cells from tail regeneration buds. High Hoechst dye efflux capacity is thought to be a common characteristic of several types of mammalian tissue stem cells; these stem cells, designated as the "side population (SP)," may be enriched by flow cytometry (SP method). To evaluate the effectiveness of stem cell enrichment using the SP method in regenerating X. laevis tadpole tails, we performed single-cell RNA sequencing (scRNA-seq) of SP cells from regeneration buds and analyzed the frequency of satellite cells, which are muscle stem/progenitor cells expressing pax7. The pax7-expressing cells were enriched in the SP compared with whole normal tails and regeneration buds. Furthermore, hes1-expressing cells, which are assumed to be neural stem/progenitor cells, were also enriched in the SP. Our findings suggest that the SP method is efficient for successfully enriching tissue stem cells in regenerating X. laevis tadpole tails, indicating that the combination of the SP method and scRNA-seq is useful for studying tissue stem cells that contribute to tail regeneration.

    DOI PubMed

  • Xenopus laevis il11ra.L is an experimentally proven interleukin-11 receptor component that is required for tadpole tail regeneration.

    Shunya Suzuki, Kayo Sasaki, Taro Fukazawa, Takeo Kubo

    Scientific reports   12 ( 1 ) 1903 - 1903  2022.02  [International journal]

    　View Summary

    Xenopus laevis tadpoles possess high regenerative ability and can regenerate functional tails after amputation. An early event in regeneration is the induction of undifferentiated cells that form the regenerated tail. We previously reported that interleukin-11 (il11) is upregulated immediately after tail amputation to induce undifferentiated cells of different cell lineages, indicating a key role of il11 in initiating tail regeneration. As Il11 is a secretory factor, Il11 receptor-expressing cells are thought to mediate its function. X. laevis has a gene annotated as interleukin 11 receptor subunit alpha on chromosome 1L (il11ra.L), a putative subunit of the Il11 receptor complex, but its function has not been investigated. Here, we show that nuclear localization of phosphorylated Stat3 induced by Il11 is abolished in il11ra.L knocked-out culture cells, strongly suggesting that il11ra.L encodes an Il11 receptor component. Moreover, knockdown of il11ra.L impaired tadpole tail regeneration, suggesting its indispensable role in tail regeneration. We also provide a model showing that Il11 functions via il11ra.L-expressing cells in a non-cell autonomous manner. These results highlight the importance of il11ra.L-expressing cells in tail regeneration.

    DOI PubMed

  • Low-temperature incubation improves both knock-in and knock-down efficiencies by the CRISPR/Cas9 system in Xenopus laevis as revealed by quantitative analysis.

    Sumika Kato, Taro Fukazawa, Takeo Kubo

    Biochemical and biophysical research communications   543   50 - 55  2021.03  [International journal]

    　View Summary

    The recent development of the CRISPR/Cas9-mediated gene editing technique has provided various gene knock-down and knock-in methods for Xenopus laevis. Gene-edited F0 individuals created by these methods, however, are mosaics with both mutated/knocked-in and unedited wild-type cells, and therefore precise determination and higher efficiency of knock-down and knock-in methods are desirable, especially for analyses of F0 individuals. To clarify the ratio of cells that are gene-edited by CRISPR/Cas9 methods to the whole cells in F0 individuals, we subjected Inference of CRISPR Edits analysis for knock-down experiments and flow cytometry for knock-in experiments to the F0 individuals. With these quantitative methods, we showed that low-temperature incubation of X. laevis embryos after microinjection improved the mutation rate in the individuals. Moreover, we applied low-temperature incubation when using a knock-in method with long single-strand DNA and found improved knock-in efficiency. Our results provide a simple and useful way to evaluate and improve the efficiency of gene editing in X. laevis.

    DOI PubMed

  • Developmental stage-specific distribution and phosphorylation of Mblk-1, a transcription factor involved in ecdysteroid-signaling in the honey bee brain

    Hitomi Kumagai, Takekazu Kunieda, Korefumi Nakamura, Yasuhiro Matsumura, Manami Namiki, Hiroki Kohno, Takeo Kubo

    Scientific Reports   10 ( 1 )  2020.12

    DOI PubMed

  • Developmental stage-specific distribution and phosphorylation of Mblk-1, a transcription factor involved in ecdysteroid-signaling in the honey bee brain

    Hitomi Kumagai, Takekazu Kunieda, Korefumi Nakamura, Yasuhiro Matsumura, Manami Namiki, Hiroki Kohno, Takeo Kubo

    Scientific Reports   10 ( 1 ) 11577 - 11577  2020.12  [Refereed]  [International journal]

    DOI PubMed

  • Neural activity mapping of bumble bee (Bombus ignitus) brains during foraging flight using immediate early genes.

    Shiori Iino, Yurika Shiota, Masakazu Nishimura, Shinichi Asada, Masato Ono, Takeo Kubo

    Scientific reports   10 ( 1 ) 7887 - 7887  2020.05  [International journal]

    　View Summary

    Honey bees and bumble bees belong to the same family (Apidae) and their workers exhibit a division of labor, but the style of division of labor differs between species. The molecular and neural bases of the species-specific social behaviors of Apidae workers have not been analyzed. Here, we focused on two immediate early genes, hormone receptor 38 (HR38) and early growth response gene-1 (Egr1), and late-upregulated ecdysone receptor (EcR), all of which are upregulated by foraging flight and expressed preferentially in the small-type Kenyon cells of the mushroom bodies (MBs) in the honey bee brain. Gene expression analyses in Bombus ignitus revealed that HR38 and Egr1, but not EcR, exhibited an immediate early response during awakening from CO2 anesthesia. Both premature mRNA for HR38 and mature mRNA for Egr1 were induced during foraging flight, and mRNAs for HR38 and Egr1 were sparsely detected inside the whole MB calyces. In contrast, EcR expression was higher in forager brains than in nurse bees and was expressed preferentially in the small-type Kenyon cells inside the MBs. Our findings suggest that Kenyon cells are active during foraging flight and that the function of late-upregulated EcR in the brain is conserved among these Apidae species.

    DOI PubMed

  • Genetics in the Honey Bee: Achievements and Prospects toward the Functional Analysis of Molecular and Neural Mechanisms Underlying Social Behaviors.

    Kohno H, Kubo T

    Insects   10 ( 10 )  2019.10  [Refereed]  [International journal]

    　View Summary

    The European honey bee is a model organism for studying social behaviors. Comprehensive analyses focusing on the differential expression profiles of genes between the brains of nurse bees and foragers, or in the mushroom bodies-the brain structure related to learning and memory, and multimodal sensory integration-has identified candidate genes related to honey bee behaviors. Despite accumulating knowledge on the expression profiles of genes related to honey bee behaviors, it remains unclear whether these genes actually regulate social behaviors in the honey bee, in part because of the scarcity of genetic manipulation methods available for application to the honey bee. In this review, we describe the genetic methods applied to studies of the honey bee, ranging from classical forward genetics to recently developed gene modification methods using transposon and CRISPR/Cas9. We then discuss future functional analyses using these genetic methods targeting genes identified by the preceding research. Because no particular genes or neurons unique to social insects have been found yet, further exploration of candidate genes/neurons correlated with sociality through comprehensive analyses of mushroom bodies in the aculeate species can provide intriguing targets for functional analyses, as well as insight into the molecular and neural bases underlying social behaviors.

    DOI PubMed

  • Detection of Phospholipase C Activity in the Brain Homogenate from the Honeybee.

    Suenami S, Miyazaki R, Kubo T

    Journal of visualized experiments : JoVE   ( 139 )  2018.09  [Refereed]  [International journal]

    　View Summary

    The honeybee is a model organism for evaluating complex behaviors and higher brain function, such as learning, memory, and division of labor. The mushroom body (MB) is a higher brain center proposed to be the neural substrate of complex honeybee behaviors. Although previous studies identified genes and proteins that are differentially expressed in the MBs and other brain regions, the activities of the proteins in each region are not yet fully understood. To reveal the functions of these proteins in the brain, pharmacologic analysis is a feasible approach, but it is first necessary to confirm that pharmacologic manipulations indeed alter the protein activity in these brain regions. We previously identified a higher expression of genes encoding phospholipase C (PLC) in the MBs than in other brain regions, and pharmacologically assessed the involvement of PLC in honeybee behavior. In that study, we biochemically tested two pharmacologic agents and confirmed that they decreased PLC activity in the MBs and other brain regions. Here, we present a detailed description of how to detect PLC activity in honeybee brain homogenate. In this assay system, homogenates derived from different brain regions are reacted with a synthetic fluorogenic substrate, and fluorescence resulting from PLC activity is quantified and compared between brain regions. We also describe our evaluation of the inhibitory effects of certain drugs on PLC activity using the same system. Although this system is likely affected by other endogenous fluorescence compounds and/or the absorbance of the assay components and tissues, the measurement of PLC activity using this system is safer and easier than that using the traditional assay, which requires radiolabeled substrates. The simple procedure and manipulations allow us to examine PLC activity in the brains and other tissues of honeybees involved in different social tasks.

    DOI PubMed

  • mKast is dispensable for normal development and sexual maturation of the male European honeybee.

    Kohno H, Kubo T

    Scientific reports   8 ( 1 ) 11877 - 11877  2018.08  [Refereed]  [International journal]

    　View Summary

    The European honeybee (Apis mellifera L.) exhibits various social behaviors. The molecular and neural mechanisms underlying these behaviors have long been explored, but causal relations between genes or neurons and behaviors remain to be elucidated because effective gene manipulation methods in the honeybee have not been available until recently. We recently established a basic technology to produce mutant honeybee drones using CRISPR/Cas9. Here we produced mutant drones using CRISPR/Cas9 targeting mKast, which is preferentially expressed in a certain subtype of class I Kenyon cells that comprise the mushroom bodies in the honeybee brain. By immunoblot analysis, we showed that mKast protein expression was completely lost in the mutant drone heads. In addition, during the production process of homozygous mutant workers, we demonstrated that heterozygous mutant workers could be produced by artificial insemination of wild-type queens with the sperm of mutant drones, indicating that mKast mutant drones were sexually mature. These results demonstrate that mKast is dispensable for normal development and sexual maturation in drone honeybees, and allow us to proceed with the production of homozygous mutant workers for the analysis of a particular gene by gene knockout in the future.

    DOI PubMed

  • Shotgun proteomics deciphered age/division of labor-related functional specification of three honeybee (Apis mellifera L.) exocrine glands

    Toshiyuki Fujita, Hiroko Kozuka-Hata, Yutaro Hori, Jun Takeuchi, Takeo Kubo, Masaaki Oyama

    PLoS ONE   13 ( 2 ) e0191344  2018.02  [Refereed]  [International journal]

    DOI PubMed

  • Pharmacologic inhibition of phospholipase C in the brain attenuates early memory formation in the honeybee (Apis mellifera L.)

    Shota Suenami, Shiori Iino, Takeo Kubo

    Biology Open   7 ( 1 )  2018  [Refereed]  [International journal]

    DOI PubMed

  • Kenyon Cell Subtypes/Populations in the Honeybee Mushroom Bodies: Possible Function Based on Their Gene Expression Profiles, Differentiation, Possible Evolution, and Application of Genome Editing.

    Suenami S, Oya S, Kohno H, Kubo T

    Frontiers in psychology   9   1717 - 1717  2018  [Refereed]  [International journal]

    　View Summary

    Mushroom bodies (MBs), a higher-order center in the honeybee brain, comprise some subtypes/populations of interneurons termed as Kenyon cells (KCs), which are distinguished by their cell body size and location in the MBs, as well as their gene expression profiles. Although the role of MBs in learning ability has been studied extensively in the honeybee, the roles of each KC subtype and their evolution in hymenopteran insects remain mostly unknown. This mini-review describes recent progress in the analysis of gene/protein expression profiles and possible functions of KC subtypes/populations in the honeybee. Especially, the discovery of novel KC subtypes/populations, the "middle-type KCs" and "KC population expressing FoxP," necessitated a redefinition of the KC subtype/population. Analysis of the effects of inhibiting gene function in a KC subtype-preferential manner revealed the function of the gene product as well as of the KC subtype where it is expressed. Genes expressed in a KC subtype/population-preferential manner can be used to trace the differentiation of KC subtypes during the honeybee ontogeny and the possible evolution of KC subtypes in hymenopteran insects. Current findings suggest that the three KC subtypes are unique characteristics to the aculeate hymenopteran insects. Finally, prospects regarding future application of genome editing for the study of KC subtype functions in the honeybee are described. Genes expressed in a KC subtype-preferential manner can be good candidate target genes for genome editing, because they are likely related to highly advanced brain functions and some of them are dispensable for normal development and sexual maturation in honeybees.

    DOI PubMed

  • Increased complexity of mushroom body Kenyon cell subtypes in the brain is associated with behavioral evolution in hymenopteran insects

    Satoyo Oya, Hiroki Kohno, Yooichi Kainoh, Masato Ono, Takeo Kubo

    SCIENTIFIC REPORTS   7 ( 1 ) 13785 - 13785  2017.10  [Refereed]  [International journal]

    DOI PubMed

  • interleukin-11 induces and maintains progenitors of different cell lineages during Xenopus tadpole tail regeneration

    Hiroshi Tsujioka, Takekazu Kunieda, Yuki Katou, Katsuhiko Shirahige, Taro Fukazawa, Takeo Kubo

    NATURE COMMUNICATIONS   8 ( 1 ) 495  2017.09  [Refereed]

    DOI PubMed

  • Gene expression and immunohistochemical analyses of mKast suggest its late pupal and adult-specific functions in the honeybee brain (vol 12, e0176809, 2017)

    Atsuhiro Yamane, Hiroki Kohno, Tsubomi Ikeda, Kumi Kaneko, Atsushi Ugajin, Toshiyuki Fujita, Takekazu Kunieda, Takeo Kubo

    PLOS ONE   12 ( 8 ) e0176809  2017.08  [Refereed]

    DOI PubMed

  • Gene expression and immunohistochemical analyses of mKast suggest its late pupal and adult-specific functions in the honeybee brain (vol 12, e0176809, 2017)

    Atsuhiro Yamane, Hiroki Kohno, Tsubomi Ikeda, Kumi Kaneko, Atsushi Ugajin, Toshiyuki Fujita, Takekazu Kunieda, Takeo Kubo

    PLOS ONE   12 ( 8 ) e0183522  2017.08  [Refereed]

    DOI PubMed

  • Gene expression and immunohistochemical analyses of mKast suggest its late pupal and adult-specific functions in the honeybee brain (vol 12, e0176809, 2017)

    Atsuhiro Yamane, Hiroki Kohno, Tsubomi Ikeda, Kumi Kaneko, Atsushi Ugajin, Toshiyuki Fujita, Takekazu Kunieda, Takeo Kubo

    PLOS ONE   12 ( 8 ) e0186041  2017.08  [Refereed]

    DOI PubMed

  • Clonal analysis of construction mechanism of the adult telencephalon mediated by post-hatch neurogenesiss in medaka fish

    Yasuko Isoe, Ryohei Nakamura, Yasuhiro Kamei, Shigenori Nonaka, Teruhiro Okuyama, Atsushi Shimizu, Takeo Kubo, Hiroyuki Takeda, Hideaki Takeuchi

    MECHANISMS OF DEVELOPMENT   145   S118 - S119  2017.07

    DOI

  • Gene expression and immunohistochemical analyses of mKast suggest its late pupal and adult-specific functions in the honeybee brain

    Atsuhiro Yamane, Hiroki Kohno, Tsubomi Ikeda, Kumi Kaneko, Atsushi Ugajin, Toshiyuki Fujita, Takekazu Kunieda, Takeo Kubo

    PLOS ONE   12 ( 5 ) e0176809  2017.05  [Refereed]

    DOI PubMed

  • Mblk-1 Transcription Factor Family: Its Roles in Various Animals and Regulation by NOL4 Splice Variants in Mammals

    Seika Takayanagi-Kiya, Taketoshi Kiya, Takekazu Kunieda, Takeo Kubo

    INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES   18 ( 2 )  2017.02  [Refereed]

    DOI PubMed

  • メダカにおいてバソトシン系は配偶者防衛行動を制御する(Vasotocin system regulates mate-guarding behavior in medaka fish)

    Yokoi Saori, Okuyama Teruhiro, Ansai Satoshi, Kamei Yasuhiro, Taniguchi Yoshihito, Kinoshita Masato, Young Larry J., Takemori Nobuaki, Kubo Takeo, Naruse Kiyoshi, Takeuchi Hideaki

    比較生理生化学   33 ( 4 ) 149 - 149  2016.12

  • Establishment of an isogenic strain of the desiccation-sensitive tardigrade Isohypsibius myrops (Parachela, Eutardigrada) and its life history traits

    Makiko Ito, Tokiko Saigo, Wataru Abe, Takeo Kubo, Takekazu Kunieda

    ZOOLOGICAL JOURNAL OF THE LINNEAN SOCIETY   178 ( 4 ) 863 - 870  2016.12

    DOI

  • Acute phase response in amputated tail stumps and neural tissue-preferential expression in tail bud embryos of the Xenopus neuronal pentraxin I gene

    Yuko Hatta-Kobayashi, Mie Toyama-Shirai, Takehiro Yamanaka, Mayuko Takamori, Yoko Wakabayashi, Yuko Naora, Takekazu Kunieda, Taro Fukazawa, Takeo Kubo

    DEVELOPMENT GROWTH & DIFFERENTIATION   58 ( 9 ) 688 - 701  2016.12  [Refereed]

    DOI PubMed

  • Production of Knockout Mutants by CRISPR/Cas9 in the European Honeybee, Apis mellifera L.

    Hiroki Kohno, Shota Suenami, Hideaki Takeuchi, Tetsuhiko Sasaki, Takeo Kubo

    ZOOLOGICAL SCIENCE   33 ( 5 ) 505 - 512  2016.10  [Refereed]

    DOI PubMed

  • Extremotolerant tardigrade genome and improved radiotolerance of human cultured cells by tardigrade-unique protein

    Takuma Hashimoto, Daiki D. Horikawa, Yuki Saito, Hirokazu Kuwahara, Hiroko Kozuka-Hata, Tadasu Shin-, Yohei Minakuchi, Kazuko Ohishi, Ayuko Motoyama, Tomoyuki Aizu, Atsushi Enomoto, Koyuki Kondo, Sae Tanaka, Yuichiro Hara, Shigeyuki Koshikawa, Hiroshi Sagara, Toru Miura, Shin-ichi Yokobori, Kiyoshi Miyagawa, Yutaka Suzuki, Takeo Kubo, Masaaki Oyama, Yuji Kohara, Asao Fujiyama, Kazuharu Arakawa, Toshiaki Katayama, Atsushi Toyoda, Takekazu Kunieda

    NATURE COMMUNICATIONS   7   12808  2016.09  [Refereed]

    DOI PubMed

  • Preparation of Single-cohort Colonies and Hormone Treatment of Worker Honeybees to Analyze Physiology Associated with Role and/or Endocrine System

    Takayuki Ueno, Kiyoshi Kawasaki, Takeo Kubo

    JOVE-JOURNAL OF VISUALIZED EXPERIMENTS   ( 115 )  2016.09  [Refereed]

    DOI PubMed

  • Ontogeny and Sexual Differences in Swimming Proximity to Conspecifics in Response to Visual Cues in Medaka Fish

    Yasuko Isoe, Yumi Konagaya, Saori Yokoi, Takeo Kubo, Hideaki Takeuchi

    ZOOLOGICAL SCIENCE   33 ( 3 ) 246 - 254  2016.06  [Refereed]

    DOI PubMed

  • Analysis of the Differentiation of Kenyon Cell Subtypes Using Three Mushroom Body-Preferential Genes during Metamorphosis in the Honeybee (Apis mellifera L.)

    Shota Suenami, Rajib Kumar Paul, Hideaki Takeuchi, Genta Okude, Tomoko Fujiyuki, Kenichi Shirai, Takeo Kubo

    PLOS ONE   11 ( 6 ) e0157841  2016.06  [Refereed]

    DOI PubMed

  • Phyhd1, an XPhyH-like homologue, is induced in mouse T cells upon T cell stimulation

    Yuri Furusawa, Takeo Kubo, Taro Fukazawa

    BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS   472 ( 3 ) 551 - 556  2016.04  [Refereed]

    DOI PubMed

  • Gene expression profiles and neural activities of Kenyon cell subtypes in the honeybee brain: identification of novel 'middle-type' Kenyon cells.

    Kaneko K, Suenami S, Kubo T

    Zoological letters   2   14 - 14  2016  [Refereed]  [International journal]

    　View Summary

    In the honeybee (Apis mellifera L.), it has long been thought that the mushroom bodies, a higher-order center in the insect brain, comprise three distinct subtypes of intrinsic neurons called Kenyon cells. In class-I large-type Kenyon cells and class-I small-type Kenyon cells, the somata are localized at the edges and in the inner core of the mushroom body calyces, respectively. In class-II Kenyon cells, the somata are localized at the outer surface of the mushroom body calyces. The gene expression profiles of the large- and small-type Kenyon cells are distinct, suggesting that each exhibits distinct cellular characteristics. We recently identified a novel gene, mKast (middle-type Kenyon cell-preferential arrestin-related gene-1), which has a distinctive expression pattern in the Kenyon cells. Detailed expression analyses of mKast led to the discovery of novel 'middle-type' Kenyon cells characterized by their preferential mKast-expression in the mushroom bodies. The somata of the middle-type Kenyon cells are localized between the large- and small-type Kenyon cells, and the size of the middle-type Kenyon cell somata is intermediate between that of large- and small-type Kenyon cells. Middle-type Kenyon cells appear to differentiate from the large- and/or small-type Kenyon cell lineage(s). Neural activity mapping using an immediate early gene, kakusei, suggests that the small-type and some middle-type Kenyon cells are prominently active in the forager brain, suggesting a potential role in processing information during foraging flight. Our findings indicate that honeybee mushroom bodies in fact comprise four types of Kenyon cells with different molecular and cellular characteristics: the previously known class-I large- and small-type Kenyon cells, class-II Kenyon cells, and the newly identified middle-type Kenyon cells described in this review. As the cellular characteristics of the middle-type Kenyon cells are distinct from those of the large- and small-type Kenyon cells, their careful discrimination will be required in future studies of honeybee Kenyon cell subtypes. In this review, we summarize recent progress in analyzing the gene expression profiles and neural activities of the honeybee Kenyon cell subtypes, and discuss possible roles of each Kenyon cell subtype in the honeybee brain.

    DOI PubMed

  • Suggested Involvement of PP1/PP2A Activity and De Novo Gene Expression in Anhydrobiotic Survival in a Tardigrade, Hypsibius dujardini, by Chemical Genetic Approach

    Koyuki Kondo, Takeo Kubo, Takekazu Kunieda

    PLOS ONE   10 ( 12 ) e0144803  2015.12  [Refereed]

    DOI PubMed

  • Changes in the Gene Expression Profiles of the Hypopharyngeal Gland of Worker Honeybees in Association with Worker Behavior and Hormonal Factors

    Takayuki Ueno, Hideaki Takeuchi, Kiyoshi Kawasaki, Takeo Kubo

    PLOS ONE   10 ( 6 ) e0130206  2015.06  [Refereed]

    DOI PubMed

  • Unique Gene Expression Profile of the Proliferating Xenopus Tadpole Tail Blastema Cells Deciphered by RNA-Sequencing Analysis

    Hiroshi Tsujioka, Takekazu Kunieda, Yuki Katou, Katsuhiko Shirahige, Takeo Kubo

    PLOS ONE   10 ( 3 ) e0111655  2015.03  [Refereed]

    DOI PubMed

  • Novel Mitochondria-Targeted Heat-Soluble Proteins Identified in the Anhydrobiotic Tardigrade Improve Osmotic Tolerance of Human Cells

    Sae Tanaka, Junko Tanaka, Yoshihiro Miwa, Daiki D. Horikawa, Toshiaki Katayama, Kazuharu Arakawa, Atsushi Toyoda, Takeo Kubo, Takekazu Kunieda

    PLOS ONE   10 ( 2 ) e0118272  2015.02  [Refereed]

    DOI PubMed

  • A new antigenic marker specifically labels a subpopulation of the class II Kenyon cells in the brain of the European honeybee Apis mellifera

    Takayuki Watanabe, Takeo Kubo

    Biophysics (Japan)   11   73 - 77  2015  [Refereed]

    DOI PubMed

  • An essential role of the arginine vasotocin system in mate-guarding behaviors in triadic relationships of medaka fish (Oryzias latipes).

    Saori Yokoi, Teruhiro Okuyama, Yasuhiro Kamei, Kiyoshi Naruse, Yoshihito Taniguchi, Satoshi Ansai, Masato Kinoshita, Larry J Young, Nobuaki Takemori, Takeo Kubo, Hideaki Takeuchi

    PLoS genetics   11 ( 2 ) e1005009  2015  [Refereed]  [International journal]

    　View Summary

    To increase individual male fitness, males of various species remain near a (potential) mating partner and repel their rivals (mate-guarding). Mate-guarding is assumed to be mediated by two different types of motivation: sexual motivation toward the opposite sex and competitive motivation toward the same sex. The genetic/molecular mechanisms underlying how mate presence affects male competitive motivation in a triadic relationship has remained largely unknown. Here we showed that male medaka fish prominently exhibit mate-guarding behavior. The presence of a female robustly triggers male-male competition for the female in a triadic relationship (2 males and 1 female). The male-male competition resulted in one male occupying a dominant position near the female while interfering with the other male's approach of the female. Paternity testing revealed that the dominant male had a significantly higher mating success rate than the other male in a triadic relationship. We next generated medaka mutants of arginine-vasotocin (avt) and its receptors (V1a1, V1a2) and revealed that two genes, avt and V1a2, are required for normal mate-guarding behavior. In addition, behavioral analysis of courtship behaviors in a dyadic relationship and aggressive behaviors within a male group revealed that avt mutant males displayed decreased sexual motivation but showed normal aggression. In contrast, heterozygote V1a2 mutant males displayed decreased aggression, but normal mate-guarding and courtship behavior. Thus, impaired mate-guarding in avt and V1a2 homozygote mutants may be due to the loss of sexual motivation toward the opposite sex, and not to the loss of competitive motivation toward rival males. The different behavioral phenotypes between avt, V1a2 heterozygote, and V1a2 homozygote mutants suggest that there are redundant systems to activate V1a2 and that endogenous ligands activating the receptor may differ according to the social context.

    DOI PubMed

  • Splicing Variants of NOL4 Differentially Regulate the Transcription Activity of Mlr1 and Mlr2 in Cultured Cells

    Seika Takayanagi-Kiya, Kayo Misawa-Hojo, Taketoshi Kiya, Takekazu Kunieda, Takeo Kubo

    ZOOLOGICAL SCIENCE   31 ( 11 ) 735 - 740  2014.11  [Refereed]

    DOI PubMed

  • A neural mechanism underlying mating preferences for familiar individuals in medaka fish.

    Teruhiro Okuyama, Saori Yokoi, Hideki Abe, Yasuko Isoe, Yuji Suehiro, Haruka Imada, Minoru Tanaka, Takashi Kawasaki, Shunsuke Yuba, Yoshihito Taniguchi, Yasuhiro Kamei, Kataaki Okubo, Atsuko Shimada, Kiyoshi Naruse, Hiroyuki Takeda, Yoshitaka Oka, Takeo Kubo, Hideaki Takeuchi

    Science (New York, N.Y.)   343 ( 6166 ) 91 - 4  2014.01  [Refereed]  [International journal]

    　View Summary

    Social familiarity affects mating preference among various vertebrates. Here, we show that visual contact of a potential mating partner before mating (visual familiarization) enhances female preference for the familiarized male, but not for an unfamiliarized male, in medaka fish. Terminal-nerve gonadotropin-releasing hormone 3 (TN-GnRH3) neurons, an extrahypothalamic neuromodulatory system, function as a gate for activating mating preferences based on familiarity. Basal levels of TN-GnRH3 neuronal activity suppress female receptivity for any male (default mode). Visual familiarization facilitates TN-GnRH3 neuron activity (preference mode), which correlates with female preference for the familiarized male. GnRH3 peptides, which are synthesized specifically in TN-GnRH3 neurons, are required for the mode-switching via self-facilitation. Our study demonstrates the central neural mechanisms underlying the regulation of medaka female mating preference based on visual social familiarity.

    DOI PubMed

  • Genetic control of startle behavior in medaka fish.

    Satomi Tsuboko, Tetsuaki Kimura, Minori Shinya, Yuji Suehiro, Teruhiro Okuyama, Atsuko Shimada, Hiroyuki Takeda, Kiyoshi Naruse, Takeo Kubo, Hideaki Takeuchi

    PloS one   9 ( 11 ) e112527  2014  [Refereed]  [International journal]

    　View Summary

    Genetic polymorphisms are thought to generate intraspecific behavioral diversities, both within and among populations. The mechanisms underlying genetic control of behavioral properties, however, remain unclear in wild-type vertebrates, including humans. To explore this issue, we used diverse inbred strains of medaka fish (Oryzias latipes) established from the same and different local populations. Medaka exhibit a startle response to a visual stimulus (extinction of illumination) by rapidly bending their bodies (C-start) 20-ms after the stimulus presentation. We measured the rates of the response to repeated stimuli (1-s interval, 40 times) among four inbred strains, HNI-I, HNI-II, HO5, and Hd-rR-II1, and quantified two properties of the startle response: sensitivity (response rate to the first stimulus) and attenuation of the response probability with repeated stimulus presentation. Among the four strains, the greatest differences in these properties were detected between HNI-II and Hd-rR-II1. HNI-II exhibited high sensitivity (approximately 80%) and no attenuation, while Hd-rR-II1 exhibited low sensitivity (approximately 50%) and almost complete attenuation after only five stimulus presentations. Our findings suggested behavioral diversity of the startle response within a local population as well as among different populations. Linkage analysis with F2 progeny between HNI-II and Hd-rR-II1 detected quantitative trait loci (QTL) highly related to attenuation, but not to sensitivity, with a maximum logarithm of odds score of 11.82 on linkage group 16. The three genotypes (homozygous for HNI-II and Hd-rR-II1 alleles, and heterozygous) at the marker nearest the QTL correlated with attenuation. Our findings are the first to suggest that a single genomic region might be sufficient to generate individual differences in startle behavior between wild-type strains. Further identification of genetic polymorphisms that define the behavioral trait will contribute to our understanding of the neural mechanisms underlying behavioral diversity, allowing us to investigate the adaptive significance of intraspecific behavioral polymorphisms of the startle response.

    DOI PubMed

  • Analysis of the molecular and neural bases underlying the honeybee 'waggle dance'

    Kumi Kaneko, Takeo Kubo

    Seikagaku   86 ( 5 ) 588 - 594  2014  [Refereed]

    PubMed

  • Correction: A New Data-Mining Method to Search for Behavioral Properties That Induce Alignment and Their Involvement in Social Learning in Medaka Fish (Oryzias Latipes).

    Ochiai T, Suehiro Y, Nishinari K, Kubo T, Takeuchi H

    PloS one   8  2013.11  [Refereed]

    DOI PubMed

  • Correction: Novel Middle-Type Kenyon Cells in the Honeybee Brain Revealed by Area-Preferential Gene Expression Analysis.

    Kaneko K, Ikeda T, Nagai M, Hori S, Umatani C, Tadano H, Ugajin A, Nakaoka T, Paul RK, Fujiyuki T, Shirai K, Kunieda T, Takeuchi H, Kubo T

    PloS one   8  2013.11  [Refereed]

    DOI PubMed

  • Identification and characterization of an Egr ortholog as a neural immediate early gene in the European honeybee (Apis mellifera L.)

    Atsushi Ugajin, Takekazu Kunieda, Takeo Kubo

    FEBS LETTERS   587 ( 19 ) 3224 - 3230  2013.10  [Refereed]

    DOI PubMed

  • A New Data-Mining Method to Search for Behavioral Properties That Induce Alignment and Their Involvement in Social Learning in Medaka Fish (Oryzias Latipes)

    Takashi Ochiai, Yuji Suehiro, Katsuhiro Nishinari, Takeo Kubo, Hideaki Takeuchi

    PLOS ONE   8 ( 9 ) e71685  2013.09  [Refereed]

    DOI PubMed

  • Novel Middle-Type Kenyon Cells in the Honeybee Brain Revealed by Area-Preferential Gene Expression Analysis

    Kumi Kaneko, Tsubomi Ikeda, Mirai Nagai, Sayaka Hori, Chie Umatani, Hiroto Tadano, Atsushi Ugajin, Takayoshi Nakaoka, Rajib Kumar Paul, Tomoko Fujiyuki, Kenichi Shirai, Takekazu Kunieda, Hideaki Takeuchi, Takeo Kubo

    PLOS ONE   8 ( 8 ) e71732  2013.08  [Refereed]

    DOI PubMed

  • Ecdysteroids and honeybee social behaviors

    Yurika Shiota, Takeo Kubo

    Hormones and Behavior     135 - 156  2013.04

  • Expression analysis of XPhyH-like during development and tail regeneration in Xenopus tadpoles: Possible role of XPhyH-like expressing immune cells in impaired tail regenerative ability

    Yuko Naora, Yuko Hishida, Taro Fukazawa, Takekazu Kunieda, Takeo Kubo

    BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS   431 ( 2 ) 152 - 157  2013.02  [Refereed]

    DOI PubMed

  • Mushroom body-preferential expression of proteins/genes involved in endoplasmic reticulum Ca2+-transport in the worker honeybee (Apis mellifera L.) brain

    Y. Uno, T. Fujiyuki, M. Morioka, T. Kubo

    INSECT MOLECULAR BIOLOGY   22 ( 1 ) 52 - 61  2013.02  [Refereed]

    DOI PubMed

  • Proteomic Analysis of the Royal Jelly and Characterization of the Functions of its Derivation Glands in the Honeybee

    Toshiyuki Fujita, Hiroko Kozuka-Hata, Hiroko Ao-Kondo, Takekazu Kunieda, Masaaki Oyama, Takeo Kubo

    JOURNAL OF PROTEOME RESEARCH   12 ( 1 ) 404 - 411  2013.01  [Refereed]

    DOI PubMed

  • Controlled Cre/loxP site-specific recombination in the developing brain in medaka fish, Oryzias latipes.

    Teruhiro Okuyama, Yasuko Isoe, Masahito Hoki, Yuji Suehiro, Genki Yamagishi, Kiyoshi Naruse, Masato Kinoshita, Yasuhiro Kamei, Atushi Shimizu, Takeo Kubo, Hideaki Takeuchi

    PloS one   8 ( 6 ) e66597  2013  [Refereed]  [International journal]

    　View Summary

    BACKGROUND: Genetic mosaic techniques have been used to visualize and/or genetically modify a neuronal subpopulation within complex neural circuits in various animals. Neural populations available for mosaic analysis, however, are limited in the vertebrate brain. METHODOLOGY/PRINCIPAL FINDINGS: To establish methodology to genetically manipulate neural circuits in medaka, we first created two transgenic (Tg) medaka lines, Tg (HSP:Cre) and Tg (HuC:loxP-DsRed-loxP-GFP). We confirmed medaka HuC promoter-derived expression of the reporter gene in juvenile medaka whole brain, and in neuronal precursor cells in the adult brain. We then demonstrated that stochastic recombination can be induced by micro-injection of Cre mRNA into Tg (HuC:loxP-DsRed-loxP-GFP) embryos at the 1-cell stage, which allowed us to visualize some subpopulations of GFP-positive cells in compartmentalized regions of the telencephalon in the adult medaka brain. This finding suggested that the distribution of clonally-related cells derived from single or a few progenitor cells was restricted to a compartmentalized region. Heat treatment of Tg(HSP:Cre x HuC:loxP-DsRed-loxP-GFP) embryos (0-1 day post fertilization [dpf]) in a thermalcycler (39°C) led to Cre/loxP recombination in the whole brain. The recombination efficiency was notably low when using 2-3 dpf embyos compared with 0-1 dpf embryos, indicating the possibility of stage-dependent sensitivity of heat-inducible recombination. Finally, using an infrared laser-evoked gene operator (IR-LEGO) system, heat shock induced in a micro area in the developing brains led to visualization of clonally-related cells in both juvenile and adult medaka fish. CONCLUSIONS/SIGNIFICANCE: We established a noninvasive method to control Cre/loxP site-specific recombination in the developing nervous system in medaka fish. This method will broaden the neural population available for mosaic analyses and allow for lineage tracing of the vertebrate nervous system in both juvenile and adult stages.

    DOI PubMed

  • Identification of kakusei, a Nuclear Non-Coding RNA, as an Immediate Early Gene from the Honeybee, and Its Application for Neuroethological Study

    Taketoshi Kiya, Atsushi Ugajin, Takekazu Kunieda, Takeo Kubo

    INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES   13 ( 12 ) 15496 - 15509  2012.12  [Refereed]

    DOI PubMed

  • Two Novel Heat-Soluble Protein Families Abundantly Expressed in an Anhydrobiotic Tardigrade

    Ayami Yamaguchi, Sae Tanaka, Shiho Yamaguchi, Hirokazu Kuwahara, Chizuko Takamura, Shinobu Imajoh-Ohmi, Daiki D. Horikawa, Atsushi Toyoda, Toshiaki Katayama, Kazuharu Arakawa, Asao Fujiyama, Takeo Kubo, Takekazu Kunieda

    PLOS ONE   7 ( 8 ) e44209  2012.08  [Refereed]

    DOI PubMed

  • p53 Mutation suppresses adult neurogenesis in medaka fish (Oryzias latipes).

    Yasuko Isoe, Teruhiro Okuyama, Yoshihito Taniguchi, Takeo Kubo, Hideaki Takeuchi

    Biochemical and biophysical research communications   423 ( 4 ) 627 - 31  2012.07  [Refereed]  [International journal]

    　View Summary

    Tumor suppressor p53 negatively regulates self-renewal of neural stem cells in the adult murine brain. Here, we report that the p53 null mutation in medaka fish (Oryzias latipes) suppressed neurogenesis in the telencephalon, independent of cell death. By using 5-bromo-29-deoxyuridine (BrdU) immunohistochemistry, we identified 18 proliferation zones in the brains of young medaka fish; in situ hybridization showed that p53 was expressed selectively in at least 12 proliferation zones. We also compared the number of BrdU-positive cells present in the whole telencephalon of wild-type (WT) and p53 mutant fish. Immediately after BrdU exposure, the number of BrdU-positive cells did not differ significantly between them. One week after BrdU-exposure, the BrdU-positive cells migrated from the proliferation zone, which was accompanied by an increased number in the WT brain. In contrast, no significant increase was observed in the p53 mutant brain. Terminal deoxynucleotidyl transferase (dUTP) nick end-labeling revealed that there was no significant difference in the number of apoptotic cells in the telencephalon of p53 mutant and WT medaka, suggesting that the decreased number of BrdU-positive cells in the mutant may be due to the suppression of proliferation rather than the enhancement of neural cell death. These results suggest that p53 positively regulates neurogenesis via cell proliferation.

    DOI PubMed

  • Detection of Neural Activity in the Brains of Japanese Honeybee Workers during the Formation of a "Hot Defensive Bee Ball"

    Atsushi Ugajin, Taketoshi Kiya, Takekazu Kunieda, Masato Ono, Tadaharu Yoshida, Takeo Kubo

    PLOS ONE   7 ( 3 ) e32902  2012.03  [Refereed]

    DOI PubMed

  • Ecdysteroid biosynthesis in workers of the European honeybee Apis mellifera L.

    Yurika Yamazaki, Makoto Kiuchi, Hideaki Takeuchi, Takeo Kubo

    INSECT BIOCHEMISTRY AND MOLECULAR BIOLOGY   41 ( 5 ) 283 - 293  2011.05  [Refereed]

    DOI PubMed CiNii

  • Dance Type and Flight Parameters Are Associated with Different Mushroom Body Neural Activities in Worker Honeybee Brains

    Taketoshi Kiya, Takeo Kubo

    PLOS ONE   6 ( 4 ) e19301  2011.04  [Refereed]

    DOI PubMed

  • Induction of c-fos transcription in the medaka brain (Oryzias latipes) in response to mating stimuli.

    Teruhiro Okuyama, Yuji Suehiro, Haruka Imada, Atsuko Shimada, Kiyoshi Naruse, Hiroyuki Takeda, Takeo Kubo, Hideaki Takeuchi

    Biochemical and biophysical research communications   404 ( 1 ) 453 - 7  2011.01  [Refereed]  [International journal]

    　View Summary

    Immediate-early genes (IEGs) are useful for mapping active brain regions in various vertebrates. Here we identified a c-fos homologue gene in medaka and demonstrated that the amounts of c-fos transcripts and proteins in the medaka brain increased in relation to an artificially evoked seizure, suggesting that the homologue gene has the characteristics of IEGs, which are used as markers of neural activity. Next, quantitative reverse-transcription-polymerase chain reaction revealed that female mating behaviors upregulated c-fos transcription in some brain regions including the telencephalon, optic tectum, and cerebellum. In addition, we performed in situ hybridization with a c-fos intron probe to detect the de novo synthesis of c-fos transcripts and confirmed induction of c-fos transcription in these brain regions after mating. This is the first report of IEG induction in response to mating stimuli in teleost fish. Our results indicated that c-fos expression was induced in response to behavioral stimuli in the medaka brain and that medaka c-fos could be a useful marker of neural activity.

    DOI PubMed

  • Expression of two microRNAs, ame-mir-276 and-1000, in the adult honeybee (Apis mellifera) brain

    Sayaka Hori, Kumi Kaneko, Takeshi H. Saito, Hideaki Takeuchi, Takeo Kubo

    APIDOLOGIE   42 ( 1 ) 89 - 102  2011.01

    DOI

  • p53 mutation caused suppression of neurogenesis in the juvenile medaka fish brain

    Yasuko Isoe, Teruhiro Okuyama, Yoshihito Taniguchi, Takeo Kubo, Hideaki Takeuchi

    NEUROSCIENCE RESEARCH   71   E239 - E240  2011

    DOI

  • Neural mechanism for female mating preference of medaka mediated by visual information

    Teruhiro Okuyama, Yuji Suehiro, Haruka Imada, Hideki Abe, Atsuko Shimada, Minoru Tanaka, Kiyoshi Naruse, Hiroyuki Takeda, Yoshitaka Oka, Takeo Kubo, Hideaki Takeuchi

    NEUROSCIENCE RESEARCH   71   E267 - E267  2011

    DOI

  • Neural basis underlying sexual selection in medaka fish

    Hideaki Takeuchi, Saori Yokoi, Yuji Suehiro, Haruka Imada, Minoru Tanaka, Takashi Kawasaki, Atsuko Shimada, Hideki Abe, Kiyoshi Naruse, Hiroyuki Takeda, Takeo Kubo, Teruhiro Okuyama

    NEUROSCIENCE RESEARCH   71   E24 - E24  2011

    DOI

  • Analysis of neural mechanism for mate-guarding behavior, using small fish medaka

    Saori Yokoi, Teruhiro Okuyama, Hideaki Takeuchi, Takeo Kubo

    NEUROSCIENCE RESEARCH   71   E265 - E265  2011

    DOI

  • Transient and permanent gene transfer into the brain of the teleost fish medaka (Oryzias latipes) using human adenovirus and the Cre-loxP system.

    Yuji Suehiro, Masato Kinoshita, Teruhiro Okuyama, Atsuko Shimada, Kiyoshi Naruse, Hiroyuki Takeda, Takeo Kubo, Mitsuhiro Hashimoto, Hideaki Takeuchi

    FEBS letters   584 ( 16 ) 3545 - 9  2010.08  [Refereed]  [International journal]

    　View Summary

    In this study, we demonstrated that human type-5 adenovirus infected the brain of the teleost fish, medaka (Oryzias latipes), in vivo. Injection of adenoviral vector into the mesencephalic ventricle of medaka larvae induced the expression of reporter genes in some parts of the telencephalon, the periventricular area of the mesencephalon and diencephalon, and the cerebellum. Additionally, the Cre-loxP system works in medaka brains using transgenic medaka carrying a vector containing DsRed2, flanked by loxP sites under control of the beta-actin promoter and downstream promoterless enhanced green fluorescent protein (EGFP). We demonstrated that the presence of green fluorescence depended on injection of adenoviral vector expressing the Cre gene and confirmed that EGFP mRNA was transcribed in the virus-injected larvae.

    DOI PubMed

  • Analysis of the mechanisms that determine tail regenerative ability in Xenopus laevis tadpoles

    Yuko Naora, Kota Kaneko, Yuko Hishida, Taro Fukazawa, Takekazu Kunieda, Takeo Kubo

    DEVELOPMENTAL BIOLOGY   344 ( 1 ) 519 - 519  2010.08

    DOI

  • Functional analysis of the honeybee (Apis mellzfera L.) salivary system using proteomics

    Toshiyuki Fujita, Hiroko Kozuka-Hata, Yuko Uno, Kenji Nishikori, Mizue Morioka, Masaaki Oyama, Takeo Kubo

    BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS   397 ( 4 ) 740 - 744  2010.07  [Refereed]

    DOI PubMed

  • Coordinated and cohesive movement of two small conspecific fish induced by eliciting a simultaneous optomotor response.

    Haruka Imada, Masahito Hoki, Yuji Suehiro, Teruhiro Okuyama, Daisuke Kurabayashi, Atsuko Shimada, Kiyoshi Naruse, Hiroyuki Takeda, Takeo Kubo, Hideaki Takeuchi

    PloS one   5 ( 6 ) e11248  2010.06  [Refereed]  [International journal]

    　View Summary

    BACKGROUND: In animal groups such as herds, schools, and flocks, a certain distance is maintained between adjacent individuals, allowing them to move as a cohesive unit. Proximate causations of the cohesive and coordinated movement under dynamic conditions, however, have been poorly understood. METHODOLOGY/PRINCIPAL FINDINGS: We established a novel and simple behavioral assay using pairs of small fish (medaka and dwarf pufferfish) by eliciting a simultaneous optomotor response (OMR). We demonstrated that two homospecific fish began to move cohesively and maintained a distance of 2 to 4 cm between them when an OMR was elicited simultaneously in the fish. The coordinated and cohesive movement was not exhibited under a static condition. During the cohesive movement, the relative position of the two fish was not stable. Furthermore, adult medaka exhibited the cohesive movement but larvae did not, despite the fact that an OMR could be elicited in larvae, indicating that this ability to coordinate movement develops during maturation. The cohesive movement was detected in homospecific pairs irrespective of body-color, sex, or albino mutation, but was not detected between heterospecific pairs, suggesting that coordinated movement is based on a conspecific interaction. CONCLUSIONS/SIGNIFICANCE: Our findings demonstrate that coordinated behavior between a pair of animals was elicited by a simultaneous OMR in two small fish. This is the first report to demonstrate induction of a schooling-like movement in a pair of fish by an OMR and to investigate the effect of age, sex, body color, and species on coordination between animals under a dynamic condition.

    DOI PubMed

  • Proliferation zones in adult medaka (Oryzias latipes) brain.

    Yoshiko Kuroyanagi, Teruhiro Okuyama, Yuji Suehiro, Haruka Imada, Atsuko Shimada, Kiyoshi Naruse, Hiroyuki Takeda, Takeo Kubo, Hideaki Takeuchi

    Brain research   1323   33 - 40  2010.04  [Refereed]  [International journal]

    　View Summary

    Cell proliferation in the adult mammalian brain is maintained at a low rate, but cell proliferation in the adult fish brain is prominent. To compare the distribution of proliferating cells among fish species, mutants, and under different growing environments, we mapped the zones of cell proliferation in the adult medaka (Oryzias latipes) brain and identified 17 proliferation zones in both male and female brains. These zones were distributed in the telencephalon (4 zones), preoptic area (2 zones), pineal body (1 zone), hypophysis (1 zone), habenular nucleus (1 zone), optic tectum (2 zones), third ventricular zone (1 zone), ventromedial nucleus (1 zone), hypothalamus (1 zone), and cerebellum (3 zones). Of the 17 zones, 16 corresponded to brain regions where cells proliferate in the zebrafish brain, suggesting that the persistence of the generation of new cells, at least in these zones, might be conserved among some fish species. We then compared the distribution of proliferation zones using two body-color mutant medaka, the T5 and Quintet, the latter of which is an albino mutant that completely lacks pigmentation. There was no apparent difference in the distribution pattern among these mutant strains. Finally, we compared these proliferation zones in the brains of isolated- and group-reared fish and detected no significant difference between the two groups. These findings demonstrate that there is persistent cell proliferation in at least these 16 zones of the adult medaka brain, irrespective of sex, body-color, and growth environment, suggesting that proliferation capacity in the 16 zones is maintained robustly in the adult medaka brain.

    DOI PubMed

  • Structural diversity and evolution of the N-terminal isoform-specific region of ecdysone receptor-A and-B1 isoforms in insects

    Takayuki Watanabe, Hideaki Takeuchi, Takeo Kubo

    BMC EVOLUTIONARY BIOLOGY   10   40  2010.02  [Refereed]

    DOI PubMed

  • In Situ Hybridization Analysis of the Expression of Futsch, Tau, and MESK2 Homologues in the Brain of the European Honeybee (Apis mellifera L.)

    Kumi Kaneko, Sayaka Hori, Mai M. Morimoto, Takayoshi Nakaoka, Rajib Kumar Paul, Tomoko Fujiyuki, Kenichi Shirai, Akiko Wakamoto, Satomi Tsuboko, Hideaki Takeuchi, Takeo Kubo

    PLOS ONE   5 ( 2 ) e9213  2010.02  [Refereed]

    DOI PubMed

  • Analysis of GABAergic and Non-GABAergic Neuron Activity in the Optic Lobes of the Forager and Re-Orienting Worker Honeybee (Apis mellifera L.)

    Taketoshi Kiya, Takeo Kubo

    PLOS ONE   5 ( 1 ) e8833  2010.01  [Refereed]

    DOI PubMed

  • Age- and division-of-labour-dependent differential expression of a novel non-coding RNA, Nb-1, in the brain of worker honeybees, Apis mellifera L.

    H. Tadano, Y. Yamazaki, H. Takeuchi, T. Kubo

    INSECT MOLECULAR BIOLOGY   18 ( 6 ) 715 - 726  2009.12  [Refereed]

    DOI PubMed

  • Distribution of Kakugo Virus and Its Effects on the Gene Expression Profile in the Brain of the Worker Honeybee Apis mellifera L.

    Tomoko Fujiyuki, Emiko Matsuzaka, Takayoshi Nakaoka, Hideaki Takeuchi, Akiko Wakamoto, Seii Ohka, Kazuhisa Sekimizu, Akio Nomoto, Takeo Kubo

    JOURNAL OF VIROLOGY   83 ( 22 ) 11560 - 11568  2009.11  [Refereed]

    DOI PubMed

  • Differential gene expression in the mandibular glands of queen and worker honeybees, Apis mellifera L.: Implications for caste-selective aldehyde and fatty acid metabolism

    Makoto Hasegawa, Satomi Asanuma, Tomoko Fujiyuki, Taketoshi Kiya, Tetsuhiko Sasaki, Daisuke Endo, Mizue Morioka, Takeo Kubo

    INSECT BIOCHEMISTRY AND MOLECULAR BIOLOGY   39 ( 10 ) 661 - 667  2009.10  [Refereed]

    DOI PubMed CiNii

  • A trophic role for Wnt-Ror kinase signaling during developmental pruning in Caenorhabditis elegans

    Yu Hayashi, Takaaki Hirotsu, Ryo Iwata, Eriko Kage-Nakadai, Hirofumi Kunitomo, Takeshi Ishihara, Yuichi Iino, Takeo Kubo

    NATURE NEUROSCIENCE   12 ( 8 ) 981 - U43  2009.08  [Refereed]

    DOI PubMed CiNii

  • Differential Gene Expression in the Hypopharyngeal Glands of Worker Honeybees (Apis mellifera L.) Associated with an Age-Dependent Role Change

    Takayuki Ueno, Takayoshi Nakaoka, Hideaki Takeuchi, Takeo Kubo

    ZOOLOGICAL SCIENCE   26 ( 8 ) 557 - 563  2009.08  [Refereed]

    DOI PubMed CiNii

  • Suppression of the immune response potentiates tadpole tail regeneration during the refractory period

    Taro Fukazawa, Yuko Naora, Takekazu Kunieda, Takeo Kubo

    DEVELOPMENT   136 ( 14 ) 2323 - 2327  2009.07  [Refereed]

    DOI PubMed

  • Morph-Dependent Expression and Subcellular Localization of Host Serine Carboxypeptidase in Bacteriocytes of the Pea Aphid Associated with Degradation of the Endosymbiotic Bacterium Buchnera

    Kenji Nishikori, Takeo Kubo, Mizue Morioka

    ZOOLOGICAL SCIENCE   26 ( 6 ) 415 - 420  2009.06  [Refereed]

    DOI PubMed CiNii

  • Age- and morph-dependent activation of the lysosomal system and Buchnera degradation in aphid endosymbiosis

    Kenji Nishikori, Kiyokazu Morioka, Takeo Kubo, Mizue Morioka

    JOURNAL OF INSECT PHYSIOLOGY   55 ( 4 ) 351 - 357  2009.04  [Refereed]

    DOI PubMed

  • Mass spectrometric map of neuropeptide expression and analysis of the gamma-prepro-tachykinin gene expression in the medaka (Oryzias latipes) brain.

    Yuji Suehiro, Akikazu Yasuda, Teruhiro Okuyama, Haruka Imada, Yoshiko Kuroyanagi, Takeo Kubo, Hideaki Takeuchi

    General and comparative endocrinology   161 ( 1 ) 138 - 45  2009.03  [Refereed]  [International journal]

    DOI PubMed

  • Inducible- and constitutive-type transcript variants of kakusei, a novel non-coding immediate early gene, in the honeybee brain

    Taketoshi Kiya, Takekazu Kunieda, Takeo Kubo

    INSECT MOLECULAR BIOLOGY   17 ( 5 ) 531 - 536  2008.10  [Refereed]

    DOI PubMed

  • [New approach toward understanding of neural basis of the honeybee "dance communication"]

    Kiya T, Hori S, Takeuchi H, Kubo T

    Tanpakushitsu kakusan koso. Protein, nucleic acid, enzyme   53 ( 11 ) 1368 - 1374  2008.09  [Refereed]  [Domestic journal]

    PubMed

  • Laying workers in queenless honeybee (Apis mellifera L.) colonies have physiological states similar to that of nurse bees but opposite that of foragers

    Takayoshi Nakaoka, Hideaki Takeuchi, Takeo Kubo

    JOURNAL OF INSECT PHYSIOLOGY   54 ( 5 ) 806 - 812  2008.05  [Refereed]

    DOI PubMed

  • Expression analysis of the FoxP homologue in the brain of the honeybee, Apis mellifera

    T. Kiya, Y. Itoh, T. Kubo

    INSECT MOLECULAR BIOLOGY   17 ( 1 ) 53 - 60  2008.02  [Refereed]

    DOI PubMed

  • The extracellular adenosine deaminase growth factor, ADGF/CECR1, plays a role in Xenopus embryogenesis via the adenosine/P1 receptor

    Ryoko Iijima, Takekazu Kunieda, Shinji Yamaguchi, Hiroko Kamigaki, Ikuko Fujii-Taira, Kazuhisa Sekimizu, Takeo Kubo, Shunji Natori, Koichi J. Homma

    JOURNAL OF BIOLOGICAL CHEMISTRY   283 ( 4 ) 2255 - 2264  2008.01  [Refereed]

    DOI PubMed CiNii

  • Analysis of the relationship between dance behavior and brain neural activity in the worker honeybee

    Taketoshi Kiya, Takeo Kubo

    NEUROSCIENCE RESEARCH   61   S112 - S112  2008

  • Molecular dissection of the brain of the honeybee social insect

    Takeo Kubo

    NEUROSCIENCE RESEARCH   61   S34 - S34  2008

  • メダカの視運動反応の情報処理過程とその神経基盤の解析

    末廣勇司, 奥山輝大, 今田はるか, 成瀬清, 島田敦子, 武田洋幸, 久保健雄, 竹内秀明

    第18回 インテリジェントシステムシンポジウム講演論文集     501 - 504  2008

  • Associative learning and discrimination of motion cues in the harnessed honeybee Apis mellifera L.

    Sayaka Hori, Hideaki Takeuchi, Takeo Kubo

    JOURNAL OF COMPARATIVE PHYSIOLOGY A-NEUROETHOLOGY SENSORY NEURAL AND BEHAVIORAL PHYSIOLOGY   193 ( 8 ) 825 - 833  2007.08  [Refereed]

    DOI PubMed

  • EcR-A expression in the brain and ovary of the honeybee (Apis mellifera L.)

    Hideaki Takeuchi, Rajib Kumar Paul, Emiko Matsuzaka, Takeo Kubo

    ZOOLOGICAL SCIENCE   24 ( 6 ) 596 - 603  2007.06  [Refereed]

    DOI PubMed CiNii

  • Identification of novel members of the xenopus Ca2+-dependent lectin family and analysis of their gene expression during tail regeneration and development

    Tomoko Ishino, Takekazu Kunieda, Shunji Natori, Kazuhisa Sekimizu, Takeo Kubo

    JOURNAL OF BIOCHEMISTRY   141 ( 4 ) 479 - 488  2007.04  [Refereed]

    DOI PubMed CiNii

  • Increased Neural Activity of a Mushroom Body Neuron Subtype in the Brains of Forager Honeybees

    Taketoshi Kiya, Takekazu Kunieda, Takeo Kubo

    PLOS ONE   2 ( 4 ) e371  2007.04  [Refereed]

    DOI PubMed

  • In vivo gene transfer into the honeybee using a nucleopolyhedrovirus vector

    Toshiya Ando, Tomoko Fujiyuki, Tomoko Kawashima, Mizue Morioka, Takeo Kubo, Haruhiko Fujiwara

    BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS   352 ( 2 ) 335 - 340  2007.01  [Refereed]

    DOI PubMed

  • Identification of proteins whose expression is up- or down-regulated in the mushroom bodies in the honeybee brain using proteomics

    Yuko Uno, Tomoko Fujiyuki, Mizue Morioka, Hideaki Takeuchi, Takeo Kubo

    FEBS LETTERS   581 ( 1 ) 97 - 101  2007.01  [Refereed]

    DOI PubMed CiNii

  • Physiological transition of honeybee workers induced by loss of the queen

    Takayoshi Nakaoka, Hideaki Takeuchi, Kenichi Shirai, Rajib Paul, Tomoko Fujiyuki, Akiko Wakamoto, Takeo Kubo

    ZOOLOGICAL SCIENCE   23 ( 12 ) 1225 - 1226  2006.12  [Refereed]

  • Prevalence and phylogeny of Kakugo virus, a novel insect picorna-like virus that infects the honeybee (Apis mellifera L.), under various colony conditions

    Tornoko Fujiyuki, Seii Ohka, Hideaki Takeuchi, Masato Ono, Akio Nomoto, Takeo Kubo

    JOURNAL OF VIROLOGY   80 ( 23 ) 11528 - 11538  2006.12  [Refereed]

    DOI PubMed

  • Expression of two ecdysteroid-regulated genes, Broad-Complex and E75, in the brain and ovary of the honeybee (Apis mellifera L.)

    Rajib Kumar Paul, Hideaki Takeuchi, Takeo Kubo

    ZOOLOGICAL SCIENCE   23 ( 12 ) 1085 - 1092  2006.12  [Refereed]

    DOI CiNii

  • Quantitative analysis of developmental processes of following behaviors in medaka using a newly established assay system

    Haruka Imada, Kenichi Shirai, Yuuji Suehiro, Kazuki Horikawa, Atsuko Shimada, Kiyoshi Naruse, Hiroyuki Takeda, Takeo Kubo, Hideaki Takeuchi

    ZOOLOGICAL SCIENCE   23 ( 12 ) 1187 - 1187  2006.12

  • Expression analysis of AmfoxP in the honeybee brain

    Taketoshi Kiya, Yuzuru Ito, Takeo Kubo

    ZOOLOGICAL SCIENCE   23 ( 12 ) 1191 - 1191  2006.12

  • Analysis of trehalose metabolism genes during anhydrobiosis in tardigrades, Milnesium tardigradum

    Takekazu Kunieda, Takeo Kubo

    ZOOLOGICAL SCIENCE   23 ( 12 ) 1197 - 1197  2006.12

  • Identification of factors that interact with Mlr1 and Mlr2: Mouse homologues of Mblk-1, a transcription factor expressed in the honeybee brain

    Kayo Misawa, Takekazu Kunieda, Takeo Kubo

    ZOOLOGICAL SCIENCE   23 ( 12 ) 1191 - 1191  2006.12

  • Analysis of mathematical principle of information processing and the molecular/neural basis that underlie optomotor response of small fishes

    Yuuji Suehiro, Haruka Imada, Kazuki Horikawa, Atsuko Shimada, Kiyoshi Naruse, Hiroyuki Takeda, Takeo Kubo, Hideaki Takeuchi

    ZOOLOGICAL SCIENCE   23 ( 12 ) 1187 - 1187  2006.12

  • Insights into social insects from the genome of the honeybee Apis mellifera

    George M. Weinstock, Gene E. Robinson, Richard A. Gibbs, Kim C. Worley, Jay D. Evans, Ryszard Maleszka, Hugh M. Robertson, Daniel B. Weaver, Martin Beye, Peer Bork, Christine G. Elsik, Klaus Hartfelder, Greg J. Hunt, Evgeny M. Zdobnov, Gro V. Amdam, Marcia M. G. Bitondi, Anita M. Collins, Alexandre S. Cristino, H. Michael G. Lattorff, Carlos H. Lobo, Robin F. A. Moritz, Francis M. F. Nunes, Robert E. Page, Zila L. P. Simoes, Diana Wheeler, Piero Carninci, Shiro Fukuda, Yoshihide Hayashizaki, Chikatoshi Kai, Jun Kawai, Naoko Sakazume, Daisuke Sasaki, Michihira Tagami, Stefan Albert, Geert Baggerman, Kyle T. Beggs, Guy Bloch, Giuseppe Cazzamali, Mira Cohen, Mark David Drapeau, Dorothea Eisenhardt, Christine Emore, Michael A. Ewing, Susan E. Fahrbach, Sylvain Foret, Cornelis J. P. Grimmelikhuijzen, Frank Hauser, Amanda B. Hummon, Jurgen Huybrechts, Andrew K. Jones, Tatsuhiko Kadowaki, Noam Kaplan, Robert Kucharski, Gerard Leboulle, Michal Linial, J. Troy Littleton, Alison R. Mercer, Timothy A. Richmond, Sandra L. Rodriguez-Zas, Elad B. Rubin, David B. Sattelle, David Schlipalius, Liliane Schoofs, Yair Shemesh, Jonathan V. Sweedler, Rodrigo Velarde, Peter Verleyen, Evy Vierstraete, Michael R. Williamson, Seth A. Ament, Susan J. Brown, Miguel Corona, Peter K. Dearden, W. Augustine Dunn, Michelle M. Elekonich, Tomoko Fujiyuki, Irene Gattermeier, Tanja Gempe, Martin Hasselmann, Tatsuhiko Kadowaki, Eriko Kage, Azusa Kamikouchi, Takeo Kubo, Robert Kucharski, Takekazu Kunieda, Marce D. Lorenzen, Natalia V. Milshina, Mizue Morioka, Kazuaki Ohashi, Ross Overbeek, Christian A. Ross, Morten Schioett, Teresa Shippy, Hideaki Takeuchi, Amy L. Toth, Judith H. Willis, Megan J. Wilson, Karl H. J. Gordon, Ivica Letunic, Kevin Hackett, Jane Peterson, Adam Felsenfeld, Mark Guyer, Michel Solignac, Richa Agarwala, Jean Marie Cornuet, Monique Monnerot, Florence Mougel, Justin T. Reese, Dominique Vautrin, Joseph J. Gillespie, Jamie J. Cannone, Robin R. Gutell, J. Spencer Johnston, Michael B. Eisen, Venky N. Iyer, Vivek Iyer, Peter Kosarev, Aaron J. Mackey, Victor Solovyev, Alexandre Souvorov, Katherine A. Aronstein, Katarina Bilikova, Yan Ping Chen, Andrew G. Clark, Laura I. Decanini, William M. Gelbart, Charles Hetru, Dan Hultmark, Jean-Luc Imler, Haobo Jiang, Michael Kanost, Kiyoshi Kimura, Brian P. Lazzaro, Dawn L. Lopez, Jozef Simuth, Graham J. Thompson, Zhen Zou, Pieter De Jong, Erica Sodergren, Miklos Csuroes, Aleksandar Milosavljevic, Kazutoyo Osoegawa, Stephen Richards, Chung-Li Shu, Laurent Duret, Eran Elhaik, Dan Graur, Juan M. Anzola, Kathryn S. Campbell, Kevin L. Childs, Derek Collinge, Madeline A. Crosby, C. Michael Dickens, L. Sian Grametes, Christina M. Grozinger, Peter L. Jones, Mireia Jorda, Xu Ling, Beverly B. Matthews, Jonathan Miller, Craig Mizzen, Miguel A. Peinado, Jeffrey G. Reid, Susan M. Russo, Andrew J. Schroeder, Susan E. St Pierre, Ying Wang, Pinglei Zhou, Huaiyang Jiang, Paul Kitts, Barbara Ruef, Anand Venkatraman, Lan Zhang, Gildardo Aquino-Perez, Charles W. Whitfield, Susanta K. Behura, Stewart H. Berlocher, Walter S. Sheppard, Deborah R. Smith, Andrew V. Suarez, Neil D. Tsutsui, Xuehong Wei, David Wheeler, Paul Havlak, Bingshan Li, Yue Liu, Erica Sodergren, Angela Jolivet, Sandra Lee, Lynne V. Nazareth, Ling-Ling Pu, Rachel Thorn, Viktor Stolc, Thomas Newman, Manoj Samanta, Waraporn A. Tongprasit, Charles Claudianos, May R. Berenbaum, Sunita Biswas, Dirk C. de Graaf, Rene Feyereisen, Reed M. Johnson, John G. Oakeshott, Hilary Ranson, Mary A. Schuler, Donna Muzny, Joseph Chacko, Clay Davis, Huyen Dinh, Rachel Gill, Judith Hernandez, Sandra Hines, Jennifer Hume, LaRonda Jackson, Christie Kovar, Lora Lewis, George Miner, Margaret Morgan, Ngoc Nguyen, Geoffrey Okwuonu, Heidi Paul, Jireh Santibanez, Glenford Savery, Amanda Svatek, Donna Villasana, Rita Wright

    NATURE   443 ( 7114 ) 931 - 949  2006.10

    DOI

  • Carbohydrate metabolism genes and pathways in insects: insights from the honey bee genome

    T. Kunieda, T. Fujiyuki, R. Kucharski, S. Foret, S. A. Ament, A. L. Toth, K. Ohashi, H. Takeuchi, A. Kamikouchi, E. Kage, M. Morioka, M. Beye, T. Kubo, G. E. Robinson, R. Maleszka

    INSECT MOLECULAR BIOLOGY   15 ( 5 ) 563 - 576  2006.10  [Refereed]

    DOI PubMed

  • Hundreds of flagellar basal bodies cover the cell surface of the endosymbiotic bacterium Buchnera aphidicola sp strain APS

    Kazuki Maezawa, Shuji Shigenobu, Hisaaki Taniguchi, Takeo Kubo, Shin-ichi Aizawa, Mizue Morioka

    JOURNAL OF BACTERIOLOGY   188 ( 18 ) 6539 - 6543  2006.09  [Refereed]

    DOI PubMed CiNii

  • Associative visual learning, color discrimination, and chromatic adaptation in the harnessed honeybee Apis mellifera L.

    Sayaka Hori, Hideaki Takeuchi, Kentaro Arikawa, Michiyo Kinoshita, Naoko Ichikawa, Masami Sasaki, Takeo Kubo

    JOURNAL OF COMPARATIVE PHYSIOLOGY A-NEUROETHOLOGY SENSORY NEURAL AND BEHAVIORAL PHYSIOLOGY   192 ( 7 ) 691 - 700  2006.07  [Refereed]

    DOI PubMed

  • Differential expression of HR38 in the mushroom bodies of the honeybee brain depends on the caste and division of labor

    Y Yamazaki, K Shirai, RK Paul, T Fujiyuki, A Wakamoto, H Takeuchi, T Kubo

    FEBS LETTERS   580 ( 11 ) 2667 - 2670  2006.05  [Refereed]

    DOI PubMed CiNii

  • Analysis of the significance and mechanism of neurite elimination using C-elegans as a model

    Yu Hayashi, Takaaki Hirotsu, Eriko Kage, Hideaki Takeuchi, Hirofumi Kunitomo, Yuichi Iino, Takeo Kubo

    NEUROSCIENCE RESEARCH   55   S185 - S185  2006

  • Analysis of the brain regions associated with the dance language of the honeybees

    Taketoshi Kiya, Takekazu Kunieda, Takeo Kubo

    NEUROSCIENCE RESEARCH   55   S95 - S95  2006

  • Analysis of the molecular mechanism of proliferating cell induction in regenerating Xenopus laevis tadpole tails

    Taro Fukazawa, Takekazu Kunieda, Takeo Kubo

    ZOOLOGICAL SCIENCE   22 ( 12 ) 1470 - 1470  2005.12

  • The small-type Kenyon cells are active in the brain of dancer honeybees

    Taketoshi Kiya, Takekazu Kunieda, Takeo Kubo

    ZOOLOGICAL SCIENCE   22 ( 12 ) 1488 - 1488  2005.12

  • Identification and analysis of trehalase from tardigrades, Milnesium tardigradum

    Takekazu Kunieda, Takeo Kubo

    ZOOLOGICAL SCIENCE   22 ( 12 ) 1493 - 1493  2005.12

  • Functional analysis of MbLK-1, a transcription factor preferentially expressed in the mushroom body of the honeybee brain

    Korefumi Nakamura, Takekazu Kunieda, Hideaki Takeuchi, Akiko Wakamoto, Takeo Kubo

    ZOOLOGICAL SCIENCE   22 ( 12 ) 1482 - 1482  2005.12

  • Identification and analysis of genes whose expression change in the brains of the workers in queenless colonies

    Takayoshi Nakaoka, Hideaki Takeuchi, Takeo Kubo

    ZOOLOGICAL SCIENCE   22 ( 12 ) 1514 - 1514  2005.12

  • Acetylcholinesterase is queen-selectively transcribed in honeybee brain and ovary

    Yuichi Takeuchi, Hideaki Takeuchi, Maya Ohara, Kenta Asahina, Takeo Kubo

    ZOOLOGICAL SCIENCE   22 ( 12 ) 1510 - 1510  2005.12

  • MBR-1, a novel helix-turn-helix transcription factor, is required for pruning excessive neurites in Caenorhabditis elegans

    E Kage, Y Hayashi, H Takeuchi, T Hirotsu, H Kunitomo, T Inoue, H Arai, Y Iino, T Kubo

    CURRENT BIOLOGY   15 ( 17 ) 1554 - 1559  2005.09  [Refereed]

    DOI PubMed

  • Kakugo virus from brains of aggressive worker honeybees

    T Fujiyuki, H Takeuchi, M Ono, S Ohka, T Sasaki, A Nomoto, T Kubo

    ADVANCES IN VIRUS RESEARCH, VOL 65   65   1 - +  2005  [Refereed]

    DOI PubMed

  • Wolbachia variant that induces two distinct reproductive phenotypes in different hosts

    Sasaki, T., Massaki, N., Kubo, T.

    Heredity   95 ( 5 )  2005

    DOI

  • Gene expression of ecdysteroid-regulated gene E74 of the honeybee in ovary and brain

    RK Paul, H Takeuchi, Y Matsuo, T Kubo

    INSECT MOLECULAR BIOLOGY   14 ( 1 ) 9 - 15  2005.01  [Refereed]

    DOI

  • [Molecular ethology using the honeybee as a model animal]

    Takeuchi H, Kunieda T, Tokuhiro-Sawata M, Park JM, Fujiyuki T, Kubo T

    Tanpakushitsu kakusan koso. Protein, nucleic acid, enzyme   49 ( 16 ) 2542 - 2548  2004.12  [Refereed]  [Domestic journal]

    PubMed

  • A novel C-elegans transcription factor MBR-1 is involved in olfactory plasticity and axon pruning

    Yu Hayashi, Eriko Kage, Takaaki Hirotsu, Hirofumi Kunitomo, Hideaki Takeuchi, Yuichi Iino, Takeo Kubo

    ZOOLOGICAL SCIENCE   21 ( 12 ) 1318 - 1318  2004.12

  • Analysis of molecular basis of associative visual learning in the honeybee Apis mellifera L.

    Sayaka Hori, Hideaki Takeuchi, Kentaro Arikawa, Michiyo Kinoshita, Masami Sasaki, Naoko Ichikawa, Takeo Kubo

    ZOOLOGICAL SCIENCE   21 ( 12 ) 1315 - 1316  2004.12

  • Attempt to map the honeybee brain regions involved in the dance language by using the immediate early gene

    Taketoshi Kiya, Takekazu Kun, Takeo Kubo

    ZOOLOGICAL SCIENCE   21 ( 12 ) 1322 - 1322  2004.12

  • Signaling in the mushroom body of the honeybee brain

    Takekazu Kunieda, Takeo Kubo

    ZOOLOGICAL SCIENCE   21 ( 12 ) 1253 - 1253  2004.12

  • Identification and characterization of a novel gene, QM1, expressed preferentially in the brain of the queen honeybee (Apis mellifera L.)

    Kenichi Shirai, Maya Ohara, Hideaki Takeuchi, Takeo Kubo

    ZOOLOGICAL SCIENCE   21 ( 12 ) 1343 - 1343  2004.12

  • Identification and analysis of the minimal promoter activity of a novel noncoding nuclear RNA gene, AncR-1, from the honeybee (Apis mellifera L.)

    M Sawata, H Takeuchi, T Kubo

    RNA-A PUBLICATION OF THE RNA SOCIETY   10 ( 7 ) 1047 - 1058  2004.07  [Refereed]

    DOI PubMed

  • Prepro-tachykinin gene expression in the brain of the honeybee Apis mellifera

    H Takeuchi, A Yasuda, Y Yasuda-Kamatani, M Sawata, Y Matsuo, A Kato, A Tsujimoto, T Nakajima, T Kubo

    CELL AND TISSUE RESEARCH   316 ( 2 ) 281 - 293  2004.05  [Refereed]

    DOI PubMed

  • Isolation and characterization of the bacteriophage WO from Wolbachia, an arthropod endosymbiont

    Y Fujii, T Kubo, H Ishikawa, T Sasaki

    BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS   317 ( 4 ) 1183 - 1188  2004.05  [Refereed]

    DOI PubMed CiNii

  • In vivo gene transfer into the adult honeybee brain by using electroporation

    T Kunieda, T Kubo

    BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS   318 ( 1 ) 25 - 31  2004.05  [Refereed]

    DOI PubMed CiNii

  • Novel insect picorna-like virus identified in the brains of aggressive worker honeybees

    T Fujiyuki, H Takeuchi, M Ono, S Ohka, T Sasaki, A Nomoto, T Kubo

    JOURNAL OF VIROLOGY   78 ( 3 ) 1093 - 1100  2004.02  [Refereed]

    DOI PubMed CiNii

  • Identification of honeybee antennal proteins/genes expressed in a sex- and/or caste selective manner

    A Kamikouchi, M Morioka, T Kubo

    ZOOLOGICAL SCIENCE   21 ( 1 ) 53 - 62  2004.01  [Refereed]

    DOI PubMed CiNii

  • Identification of a tachykinin-related neuropeptide from the honeybee brain using direct MALDI-TOF MS and its gene expression in worker, queen and drone heads

    H Takeuchi, A Yasuda, Y Yasuda-Kamatani, T Kubo, T Nakajima

    INSECT MOLECULAR BIOLOGY   12 ( 3 ) 291 - 298  2003.06  [Refereed]

    DOI

  • The activity of Mblk-1, a mushroom body-selective transcription factor from the honeybee, is modulated by the Ras/MAPK pathway

    JM Park, T Kunieda, T Kubo

    JOURNAL OF BIOLOGICAL CHEMISTRY   278 ( 20 ) 18689 - 18694  2003.05  [Refereed]

    DOI PubMed CiNii

  • Identification of genes induced in regenerating Xenopus tadpole tails by using the differential display method

    T Ishino, M Shirai, T Kunieda, K Sekimizu, S Natori, T Kubo

    DEVELOPMENTAL DYNAMICS   226 ( 2 ) 317 - 325  2003.02  [Refereed]

    DOI PubMed

  • Identification and characterization of Mlr1,2: two mouse homologues of Mblk-1, a transcription factor from the honeybee brain

    T Kunieda, JM Park, H Takeuchi, T Kubo

    FEBS LETTERS   535 ( 1-3 ) 61 - 65  2003.01  [Refereed]

    DOI PubMed CiNii

  • Interspecific transfer of Wolbachia between two lepidopteran insects expressing cytoplasmic incompatibility: A Wolbachia variant naturally infecting Cadra cautella causes male killing in Ephestia kuehniella

    T Sasaki, T Kubo, H Ishikawa

    GENETICS   162 ( 3 ) 1313 - 1319  2002.11  [Refereed]

    PubMed

  • Identification and punctate nuclear localization of a novel noncoding RNA, Ks-1, from the honeybee brain

    M Sawata, D Yoshino, H Takeuchi, A Kamikouchi, K Ohashi, T Kubo

    RNA-A PUBLICATION OF THE RNA SOCIETY   8 ( 6 ) 772 - 785  2002.06  [Refereed]

    DOI PubMed

  • DNA-binding properties of Mblk-1, a putative transcription factor from the honeybee

    JM Park, T Kunieda, H Takeuchi, T Kubo

    BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS   291 ( 1 ) 23 - 28  2002.02  [Refereed]

    DOI PubMed CiNii

  • Identification and characterization of a Drosophila homologue of ATBP

    N Aozasa, T Kubo, K Sekimizu, S Natori

    INSECT MOLECULAR BIOLOGY   11 ( 1 ) 31 - 36  2002.02  [Refereed]

    DOI PubMed

  • Identification of genes expressed preferentially in the honeybee mushroom bodies by combination of differential display and cDNA microarray

    H Takeuchi, T Fujiyuki, K Shirai, Y Matsuo, A Kamikouchi, Y Fujinawa, A Kato, A Tsujimoto, T Kubo

    FEBS LETTERS   513 ( 2-3 ) 230 - 234  2002.02  [Refereed]

    DOI PubMed CiNii

  • Identification of a novel gene, Mblk-1, that encodes a putative transcription factor expressed preferentially in the large-type Kenyon cells of the honeybee brain

    H Takeuchi, E Kage, M Sawata, A Kamikouchi, K Ohashi, M Ohara, T Fujiyuki, T Kunieda, K Sekimizu, S Natori, T Kubo

    INSECT MOLECULAR BIOLOGY   10 ( 5 ) 487 - 494  2001.10  [Refereed]

    DOI PubMed

  • Activation of the Sarcophaga lectin gene promoter by (A + T)-stretch binding protein

    Aozasa, N., Shiraishi, H., Nakanishi-Matsui, M., Kobayashi, A., Sekimizu, K., Kubo, T., Natori, S.

    European Journal of Biochemistry   268 ( 8 )  2001

    DOI

  • Interaction between ATBP and DmUbc9 in the expression of the Sarcophaga lectin gene

    Aozasa, N., Shiraishi, H., Kobayashi, A., Sekimizu, K., Kubo, T., Natori, S.

    Biochemical and Biophysical Research Communications   286 ( 5 )  2001

    DOI

  • Functional flexibility of the honey bee hypopharyngeal gland in a dequeened colony

    K Ohashi, M Sasaki, H Sasagawa, J Nakamura, S Natori, T Kubo

    ZOOLOGICAL SCIENCE   17 ( 8 ) 1089 - 1094  2000.11

    DOI PubMed CiNii

  • Concentrated expression of Ca2+/calmodulin-dependent protein kinase II and protein kinase C in the mushroom bodies of the brain of the honeybee Apis mellifera L

    A Kamikouchi, H Takeuchi, M Sawata, S Natori, T Kubo

    JOURNAL OF COMPARATIVE NEUROLOGY   417 ( 4 ) 501 - 510  2000.02  [Refereed]

    DOI

  • Gene structure and chromosome mapping of mouse transcription elongation factor S-II (Tcea1)

    T Ito, MF Seldin, MM Taketo, T Kubo, S Natori

    Gene   244 ( 1-2 ) 55 - 63  2000.02  [Refereed]

    DOI PubMed CiNii

  • Transcription elongation factor S-II confers yeast resistance to 6-azauracil by enhancing expression of the SSM1 gene

    Shimoaraiso, M., Nakanishi, T., Kubo, T., Natori, S.

    Journal of Biological Chemistry   275 ( 38 )  2000

    DOI

  • Identification, characterization and cDNA cloning of two novel proteins secreted into the external space of the regenerating leg of Periplaneta americana

    Arai, T., Kubo, T., Natori, S.

    Insect Biochemistry and Molecular Biology   30 ( 4 )  2000

    DOI

  • Participation of transcription elongation factor XSII-K1 in mesoderm-derived tissue development in Xenopus laevis

    Taira, Y., Kubo, T., Natori, S.

    Journal of Biological Chemistry   275 ( 41 )  2000

    DOI

  • Molecular cloning of cDNAs coding Sarcophaga diptericin family

    Baba, K., Kubo, T., Natori, S.

    Yakugaku Zasshi   120 ( 1 )  2000

    DOI

  • Molecular cloning and characterization of SRAM, a novel insect Rel/Ankyrin-family protein present in nuclei

    Shiraishi, H., Kobayashi, A., Sakamoto, Y., Nonaka, T., Mitsui, Y., Aozasa, N., Kubo, T., Natori, S.

    Journal of Biochemistry   127 ( 6 )  2000

    DOI

  • Molecular biology concerning to the honeybee sociality: how can we approach the evolution of insect sociality?

    Kubo, T., Ohashi, K., Sawata, M., Kamikouchi, A., Takeuchi, H.

    Tanpakushitsu kakusan koso. Protein, nucleic acid, enzyme   45 ( 7 )  2000

  • Soldier caste-specific gene expression in the mandibular glands of Hodotermopsis japonica (Isoptera : Termopsidae)

    T Miura, A Kamikouchi, M Sawata, H Takeuchi, S Natori, T Kubo, T Matsumoto

    PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA   96 ( 24 ) 13874 - 13879  1999.11  [Refereed]

    DOI PubMed CiNii

  • Expression of amylase and glucose oxidase in the hypopharyngeal gland with an age-dependent role change of the worker honeybee (Apis mellifera L.)

    K Ohashi, S Natori, T Kubo

    EUROPEAN JOURNAL OF BIOCHEMISTRY   265 ( 1 ) 127 - 133  1999.10

    DOI PubMed CiNii

  • Molecular cloning of cDNA and tissue-specific expression of the gene for SII-K1, a novel transcription elongation factor SII

    Taira, Y., Kubo, T., Natori, S.

    Genes to Cells   3 ( 5 )  1998

    DOI

  • Cloning of cDNA for regenectin, a humoral C-type lectin of Periplaneta americana, and expression of the regenectin gene during leg regeneration

    Arai, T., Kawasaki, K., Kubo, T., Natori, S.

    Insect Biochemistry and Molecular Biology   28 ( 12 )  1998

    DOI

  • Molecular cloning of cDNA for p10, a novel protein that increases in the regenerating legs of Periplaneta americana (American cockroach)

    Nomura Kitabayashi, A., Arai, T., Kubo, T., Natori, S.

    Insect Biochemistry and Molecular Biology   28 ( 10 )  1998

    DOI

  • Preferential expression of the gene for a putative inositol 1,4,5-trisphosphate receptor homologue in the mushroom bodies of the brain of the worker honeybee Apis mellifera L.

    A Kamikouchi, H Takeuchi, M Sawata, K Ohashi, S Natori, T Kubo

    BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS   242 ( 1 ) 181 - 186  1998.01  [Refereed]

    DOI PubMed CiNii

  • Change in the mode of gene expression of the hypopharyngeal gland cells with an age-dependent role change of the worker honeybee Apis mellifera L.

    K Ohashi, S Natori, T Kubo

    EUROPEAN JOURNAL OF BIOCHEMISTRY   249 ( 3 ) 797 - 802  1997.11

    DOI PubMed CiNii

  • Molecular cloning of cDNA for lysenin, a novel protein in the earthworm Eisenia foetida that causes contraction of rat vascular smooth muscle

    Sekizawa, Y., Kubo, T., Kobayashi, H., Nakajima, T., Natori, S.

    Gene   191 ( 1 )  1997

    DOI

  • Identification of the region in yeast S-II that defines species specificity in its interaction with RNA polymerase II

    Shimoaraiso, M., Nakanishi, T., Kubo, T., Natori, S.

    Journal of Biological Chemistry   272 ( 42 )  1997

    DOI

  • Molecular cloning of cDNA and analysis of expression of the gene for alpha-glucosidase from the hypopharyngeal gland of the honeybee Apis mellifera L

    K Ohashi, M Sawata, H Takeuchi, S Natori, T Kubo

    BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS   221 ( 2 ) 380 - 385  1996.04  [Refereed]

    DOI PubMed CiNii

  • Change in the expression of hypopharyngeal-gland proteins of the worker honeybees (Apis mellifera L) with age and/or role

    T Kubo, M Sasaki, J Nakamura, H Sasagawa, K Ohashi, H Takeuchi, S Natori

    JOURNAL OF BIOCHEMISTRY   119 ( 2 ) 291 - 295  1996.02  [Refereed]

  • Presence of the periplaneta lectin-related protein family in the American cockroach Periplaneta americana

    Kawasaki, K., Kubo, T., Natori, S.

    Insect Biochemistry and Molecular Biology   26 ( 4 )  1996

    DOI

  • Purification, characterization, and cDNA cloning of ABP-2 (arylphorin gene-specific binding protein-2) that specifically binds to the ABP-1-binding sequence in the arylphorin gene of Sarcophaga peregrina

    Adachi, N., Kubo, T., Natori, S.

    Journal of Biochemistry   120 ( 6 )  1996

    DOI

  • Purification and cDNA cloning of the alcohol dehydrogenase of the flesh fly Sarcophaga peregrina: A structural relationship between alcohol dehydrogenase and a 25-kDa protein

    Horio, T., Kubo, T., Natori, S.

    European Journal of Biochemistry   237 ( 3 )  1996

    DOI

  • Purification, characterization, and cDNA cloning of a galactose-specific C-type lectin from Drosophila melanogaster

    Haq, S., Kubo, T., Kurata, S., Kobayashi, A., Natori, S.

    Journal of Biological Chemistry   271 ( 33 )  1996

    DOI

  • Insect lectins and epimorphosis

    Kubo, T., Arai, T., Kawasaki, K., Natori, S.

    Trends in Glycoscience and Glycotechnology   8 ( 43 )  1996

    DOI

  • Structure-function relationship of yeast S-II in terms of stimulation of RNA polymerase II, arrest relief, and suppression of 6-azauracil sensitivity

    Nakanishi, T., Shimoaraiso, M., Kubo, T., Natori, S.

    Journal of Biological Chemistry   270 ( 15 )  1995

    DOI

  • Molecular Cloning and Nuclear Localization of ATBP, A Novel (A+T)‐Stretch‐binding Protein of Sarcophaga peregrina (Flesh Fly)

    Nakanishi?Matsui, M., Kubo, T., Natori, S.

    European Journal of Biochemistry   230 ( 2 )  1995

    DOI

  • Molecular cloning and sequencing of arylphorin-binding protein in protein granules of the Sarcophaga fat body. Implication of a post-translational processing mechanism

    Sung Ook Chung, Kubo, T., Natori, S.

    Journal of Biological Chemistry   270 ( 9 )  1995

    DOI

  • Purification and characterization of ATBP, a novel protein that binds to A/T stretches in three segments of the Sarcophaga lectin gene

    MATSUI, M., KOBAYASHI, A., KUBO, T., NATORI, S.

    European Journal of Biochemistry   219 ( 1-2 )  1994

    DOI

  • Transient appearance and localization of a 26-kDa lectin, a novel member of the periplaneta lectin family, in regenerating cockroach leg

    Kubo, T., Kawasaki, K., Natori, S.

    Developmental Biology   156 ( 2 )  1993

    DOI

  • Purification and characterization of the arylphorin gene specific binding protein from an embryonic cell line of Sarcophaga peregrina (flesh fly)

    Adachi, N., Kubo, T., Natori, S.

    Journal of Biochemistry   114 ( 1 )  1993

    DOI

  • A novel role of Periplaneta lectin as an opsonin to recognize 2-Keto-3-deoxy octonate residues of bacterial lipopolysaccharides

    Kawasaki, K., Kubo, T., Natori, S.

    Comparative Biochemistry and Physiology -- Part B: Biochemistry and   106 ( 3 )  1993

    DOI

  • Purification and characterization of a 59-kilodalton protein that specifically binds to NF-κB-binding motifs of the defense protein genes of Sarcophaga peregrina (the flesh fly)

    Kobayashi, A., Matsui, M., Kubo, T., Natori, S.

    Molecular and Cellular Biology   13 ( 7 )  1993

    DOI

  • Purification of an AU-rich RNA binding protein from Sarcophaga peregrina (flesh fly) and its identification as a thiolase

    Nanbu, R., Kubo, T., Hashimoto, T., Natori, S.

    Journal of Biochemistry   114 ( 3 )  1993

    DOI

  • Purification and characterization of a diptericin homologue from Sarcophaga peregrina (flesh fly)

    Ishikawa, M., Kubo, T., Natori, S.

    Biochemical Journal   287 ( 2 )  1992

    DOI

  • Purification and localization of p10, a novel protein that increases in nymphal regenerating legs of Periplaneta americana (American cockroach)

    Nomura, A., Kawasaki, K., Kubo, T., Natori, S.

    International Journal of Developmental Biology   36 ( 3 )  1992

  • Role of humoral lectin family in the leg regeneration of American cockroach

    Kubo, T., Kawasaki, K.

    Tanpakushitsu kakusan koso. Protein, nucleic acid, enzyme   37 ( 5 )  1992

  • Preferential deadenylation of Sarcophaga lectin mRNA during its acute phase expression

    Nanbu, R., Kubo, T., Ueno, K., Natori, S.

    BBA - Gene Structure and Expression   1088 ( 1 )  1991

    DOI

  • Localization of regenectin in regenerates of American cockroach (Periplaneta americana) legs

    Kubo, T., Kawasaki, K., Nonomura, Y., Natori, S.

    International Journal of Developmental Biology   35 ( 2 )  1991

  • Purification and characterization of lipopolysaccharide‐binding protein from hemolymph of American cockroach Periplaneta americana

    JOMORI, T., KUBO, T., NATORI, S.

    European Journal of Biochemistry   190 ( 1 )  1990

    DOI

  • Sucrose-binding lectin in regenerating cockroach (Periplaneta americana) legs: Its purification from adult hemolymph

    Kubo, T., Kawasaki, K., Natori, S.

    Insect Biochemistry   20 ( 6 )  1990

    DOI

  • Molecular Cloning of cDNA for Sarcocystatin A and Analysis of the Expression of the Sarcocystatin A Gene during Development of Sarcophaga peregrina

    Saito, H., Suzuki, T., Ueno, K., Kubo, T., Natori, S.

    Biochemistry   28 ( 4 )  1989

    DOI

  • Cloning and in vitro transcription of the Sarcophaga lectin gene

    Kobayashi, A., Hirai, H., Kubo, T., Ueno, K., Nakanishi, Y., Natori, S.

    BBA - Gene Structure and Expression   1009 ( 3 )  1989

    DOI

  • Purification and some properties of a lectin from the hemolymph of Periplaneta americana (American cockroach)

    KUBO, T., NATORI, S.

    European Journal of Biochemistry   168 ( 1 )  1987

    DOI

  • Identification of hemagglutinating protein and bactericidal activity in the hemolymph of adult Sarcophaga peregrina on injury of the body wall

    Kubo, T., Komano, H., Okada, M., Natori, S.

    Developmental and Comparative Immunology   8 ( 2 )  1984

    DOI

▼display all

Research Projects

  • Analysis of molecular bases of evolution of Kenyon cell types in the brains of hymenopteran insects.

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research

    Project Year :

    2024.04
    -
    2028.03
     

  • Analysis of molecular and neural basis of the brain correlated with behavioral characteristics of cockroaches

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research

    Project Year :

    2023.06
    -
    2026.03
     

  • Study on the molecular basis that defines organ regenerative ability in an anuran amphibian (Xenopus laevis)

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research

    Project Year :

    2020.07
    -
    2023.03
     

  • Analyses of Kenyon cell type diversification in hymenopteran insect brains and its relationship with behavioral evolution

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research

    Project Year :

    2020.04
    -
    2023.03
     

  • Study on the reactivation of regenerative ability by masking 'autoantigens' expressed in organ blastema.

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research

    Project Year :

    2014.04
    -
    2017.03
     

    Kubo Takeo, SHIRAHIGE Katsuhiko, KATOU Yuki

    　View Summary

    Xenopus laevis tadpoles transiently lose their tail regenerative ability during the ‘refractory period’, probably because immature leukocytes attack proliferating tail blastemal cells as ‘non-self’ during the period. In the present study, we aimed to clarify molecular mechanisms underlying tail regeneration through functional analyses of interleukin-11 (il-11), which is selectively expressed in the proliferating tail blastemal cells, and XPhyh-like, which is a putative marker gene for leukocytes that attack tail blastemal cells.
    The results indicated that il-11 is necessary for tail regeneration and play important roles in the induction and maintenance of undifferentiated cells of various tissue origin in tail blastema. In addition, XPhyH-like was suggested to be related to impaired tail regenerative ability caused by immature leukocytes, because murine XPhyH-like homologue (Phyhd1) was induced in leukocytes accompanied with the leukocyte activation.

  • Functional analysis of the 'middle-type' Kenyon cells that were newly identified in the honeybee brain in dance communication

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research

    Project Year :

    2014.04
    -
    2017.03
     

    Kubo Takeo

    　View Summary

    The honeybee mushroom bodies comprise 4 subtypes of class I Kenyon cells (KCs): the large-, middle- and small-type KCs, of which middle-type KCs preferentially express mKast. In the present study, we analyzed the function and evolution of the KC subtypes.
    1)mKast was suggested to be involved in various brain functions in adult honeybees. 2)We first succeeded to establish genome-editing technique in the honeybee. 3)The number of KC subtypes increased from 1 in the sawflies to 2 in the parasitic wasps and then 3 in the hornets and honeybees, suggesting its relationship with the evolution of behavioral traits in Hymenopteran insects.
    Our findings are internationally of most importance, that contribute not only to the understanding of the function of middle-type KCs but also to the progress of neurobiology in general.

  • Functional mapping of fish social brain

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research

    Project Year :

    2014.04
    -
    2017.03
     

    Takeuchi Hideaki, KAMEI Yasuhiro, Isoe Yasuko

    　View Summary

    Male medaka fish in a triad relationship (two males and one female) hold a position between the rival male and a female (mate-guarding), which results in improvement of mating success. Behavioral analysis of mutant males revealed that vasotocin system is required for dominance in mate-guarding (PLoS Genetics 2015). Our study also suggested mate-guarding behavior has an effect on visual familiarity and female mate preference. Previously we found that female medaka prefer to mate with visually-familiarized males (Science 2014). We showed that mate-guarding behaviors blocked visual familiarization of a female with the rival in a triad relationship, suggesting that female mate preference for familiar mates may help ascertain social dominance in males（Frontiers in Zoology, 2016).

  • 花粉媒介行動の脳の分子神経基盤とその生態系への影響の解明

    日本学術振興会  科学研究費助成事業

    Project Year :

    2013.04
    -
    2016.03
     

    竹内 秀明, 久保 健雄, WANG MU-YUN, WANG Mu-Yun

    　View Summary

    外国人特別研究員のWang博士はマルハナバチのコロニーの中には「注意深い(careful) 個体」と「衝動的な(impulsive) 個体」が存在することを、採餌戦略の違いから明らかにした。注意深い個体は時間をかけて蜜源を選択するが、花蜜量が多く外敵が少ない蜜源を正確に選択する。一方で衝動的な個体は短時間で蜜源を選択するが、質の高い蜜源を正確に選択できないこと(Speed-accuracy tradeoffs)を見出した（Wang et al 2015）。さらに採餌戦略における個体差が脊椎動物にもある否かを確認する目的で、Wang博士はメダカを用いて行動実験系を確立した。メダカは遺伝的背景が同一である近交系が多数、確立・維持されている。そのため近交系ごとに採餌戦略が異なることを示せれば、採餌戦略の違いは遺伝的背景の違いによると考察することができる。先行研究では、メダカに餌を与えた時に視覚刺激を同時に与えると、視覚刺激と餌の情報を連合して学習し、訓練後は視覚刺激依存に近づき行動を示すことがわかっていた（PLoS ONE 8(9): e71685, 2013）。Wang博士は、別の視覚刺激に対して電気ショックを与える実験装置を付加し、報酬と罰の両方の情報が存在する環境で、採餌行動を定量化する行動実験を確立した。Wang博士は５種類の近交系メダカの採餌行動を比較した結果、系統ごとに採餌戦略が異なる傾向があったことから、遺伝学的背景が採餌戦略を規定している可能性が考えられる。

  • アフリカツメガエルの器官再生に阻害的に働く自然炎症の分子機構の解析

    日本学術振興会  科学研究費助成事業

    Project Year :

    2012.04
    -
    2014.03
     

    久保 健雄

    　View Summary

    アフリカツメガエル（Xenopus laevis）幼生は高い尾の再生能をもつが、発育段階で一時的に再生能を失う（再生不応期）。応募者らは不応期幼生を免疫抑制すると再生能が顕著に回復することから、不応期には、再生芽細胞が未熟な免疫細胞により「非自己」として認識・攻撃されるため再生能が失われることを示唆した〔Fukazawa et al. Development (2009)〕。しかしながら、どのような種類の免疫細胞により、再生芽細胞のどの分子が「自己抗原」として認識されるのかは不明である。
    本研究では先ず、再生能を規定する免疫応答に関わる遺伝子を同定するため、ディファレンシャル・ディスプレイ法でFK506処理群（再生能あり）と無処理群（なし）の尾で発現が異なる遺伝子を検索し、新規遺伝子phytanoyl-CoA dioxygenase (XPhyH)-likeを得た。XPhyH-likeは不応期無処理群の尾の切断後に一過的に誘導され、その誘導はFK506処理で抑えられた。また、XPhyH-likeは血球分画で強く発現した。このことは、XPhyH-likeが不応期の尾切断端に一過的に浸潤し、再生を阻害する免疫細胞に発現することを示唆している〔Naora et al., Biochem. Biophys. Res. Commun. (2013)〕。
    さらに、再生芽細胞が発現する「自己抗原」を同定する目的で、尾再生芽からフローサイトメトリーを用いて核相が4nの増殖細胞を単離し、RNA-seq法により発現遺伝子を網羅的に検索した結果、尾再生芽の非増殖細胞や尾芽の増殖細胞に比べ、再生芽増殖細胞選択的に発現する遺伝子断片を23個同定した。その中には ‘Eat me not’ホモログなど「自己抗原」の候補遺伝子や、再生芽増殖細胞維持に関わると考えられるインターロイキンが含まれていた。

  • Analysis of novel proteins abundantly expressed in extremo-tolerant tardigrades.

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research

    Project Year :

    2012.04
    -
    2014.03
     

    KUNIEDA Takekazu, KUBO Takeo

    　View Summary

    Limno-terrestrial tardigrades can tolerate severe desiccation by entering an ametabolic dehydrated state called anhydrobiosis. Here, we focused and analyzed properties of novel proteins abundantly expressed in anhydrobiotc tardigrades as candidates involved in anhydrobiosis. A1 is the most abundantly expressed transcript and its protein products turned out to form an aggregated complex in a spiral shape. This aggregated structure could provide physical support to desiccating cells. We also identified a novel heat-soluble protein from less tolerant tardigrade species and showed the amount of heat-soluble proteins can correlate with tolerant ability of the species. Furthermore, as a candidate involved in protection of mitochondria against desiccation, novel mitochondrial proteins were identified.

  • Analysis of the endocrine and neural mechanisms that synergistically regulate the behavior and physiology of the worker honeybees.

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research

    Project Year :

    2011.04
    -
    2014.03
     

    KUBO Takeo, TAKEUCHI Hideaki, KUNIEDA Takekazu

    　View Summary

    In this study, we aimed to unveil brain neural mechanisms and physiological mechanisms that underlie the honeybee social behaviors. Here, we unveiled novel cellular characteristics of the large-type Kenyon cells (KCs) and identified novel 'middle-type' KCs, both of which compose the mushroom bodies, a higher brain center. The neural activities of the small-type and some 'middle-type' KCs are enhanced in the foragers, whereas those of the class II KCs are enhanced in the Ja@anese worker honeybees that are forming a 'hot defensive bee ball' against the hornets. Furthermore, we showed that the royal jelly that is secreted from young workers (nurse bees) is a cocktail of secretory proteins derived from three exocrine glands and contains cell growth factors that may affect physiological status of the other nest mates.

  • Identification and analysis of blastema-specific molecules that are targeted by immature immune system in Xenopus tadoples

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research

    Project Year :

    2010
    -
    2011
     

    KUBO Takeo

    　View Summary

    So far, we suggested that, in the' refractory period' of the Xenopus tadpoles, immature immune cells attack blastema cells of the amputated tails as' non-self', resulting in failure in tail regeneration, whereas, in the' regeneration period', regulatory T cells differentiate to reactivate tail regenerative ability. However, what kinds of immune cells target the blastema cells remains unclear. In the present study, we identified a novel gene that is supposed to be expressed in immune cells that invade amputated tail stumps to repress tail regenerative ability. This gene played an essential role in tadpole survival during the developmental stages. This gene will be available as a maker for auto-reactive immune cells that attack tail blastema cells.

  • ミツバチの視覚情報処理を支える脳のモジュール構造の分子的構築の解析

    日本学術振興会  科学研究費助成事業

    Project Year :

    2009
    -
    2010
     

    久保 健雄

    　View Summary

    ミツバチは高い視覚情報処理能力をもち、その働き蜂は巣から餌場までの距離と方向を8字ダンスにより仲間に伝達する。申請者等はミツバチ脳の"分子的解剖"を遂行し、脳の高次中枢(キノコ体)の中で、採餌飛行時の情報処理に関わると考えられる新しい"モジュール構造"に特異的に発現する遺伝子Clone #3を発見した。さらに、視覚中枢(視葉)で、明暗や物体の動きの感知に働くと考えられる"単極細胞"に選択的に発現する2つの遺伝子を発見した。本研究は、これらの既知、或いは未知のミツバチ脳の"モジュール構造"が視覚情報処理において果たす機能の解析を目的とした。
    今年度は、先の単極細胞選択的に発現する遺伝子と、視葉の腹側の頭部に水平なゾーンを形成する領域に選択的に発現する遺伝子に関して論文発表した(Kakeno et al., 2010)。さらに、働き蜂で頭部選択的に発現する2種のmiRNAを同定し、その内1種が脳領野選択的に発現することから、脳領野選択的遺伝子発現の制御に関わる可能性を示した(Hori et al., 2011)。また、ミツバチではエクダイソン情報伝達系遺伝子がキノコ体選択的に発現することを示しているが、今年度はエクダイソン合成酵素群の遺伝子発現解析と組織培養を用いたエクダイソン合成検定から、働き蜂では卵巣に加えて脳でもエクダイソンが合成されている可能性を示唆し、脳で合成されたエクダイソンが'ニューロステロイド'として脳機能調節に働く可能性を提示した(Yamazaki et al., 2011)。さらに、採餌飛行蜂の脳で活動する神経細胞を初期応答遺伝子で検出したところ、GABA陽性(抑制性)と陰性(興奮性)の両者の神経細胞が活動していることが判明し、揉餌飛行時の脳では複雑な視覚情報処理がなわれることを示唆した(Kiya et al., 2010)。

  • 昆虫をモデルとした適応戦略の分子メカニズムに関する企画調査

    日本学術振興会  科学研究費助成事業

    Project Year :

    2006
    　
    　
     

    片岡 宏誌, 上田 均, 小嶋 徹也, 久保 健雄, 藤原 晴彦, 嶋田 透

    　View Summary

    陸上において圧倒的多数の種と個体数をしめる昆虫の特質は、多様で適応的な生存戦略をそのライフサイクルに組み入れたことにある。環境変化の激しい陸上に生息する昆虫は、環境情報に対する適応戦略として、脱皮・変態・休眠などの形式を自在に変更し、大きな成功を収めた。特定領域研究(新学術領域研究)の研究領域として発展させるためには、外部情報が内分泌システムによって遺伝情報に変換される機構を現象横断的に解明し、昆虫の適応戦略に隠された共通のスキームを理解することにある。このために以下の研究調査ならびに企画を行った。
    1.研究企画会議:
    (1)06年5月29日、8月9日(東京大学農学部);片岡、藤原、久保、嶋田
    (2)06月8/9(東京大学農学部);片岡、藤原、久保、嶋田
    (3)06年7/19(東京大学理学部);片岡、嶋田、久保
    (4)07年12/26(神戸大学理学部);片岡、久保、分担者以外に尾崎(神戸大)富岡(岡山大)
    2.国外の研究状況調査:
    (1)エクジソンワークショップ06年7/9-14(ベルギー);片岡、藤原が参加、研究動向を調査
    (2)フランスパリ大学07年6/23-27;片岡が視察、研究動向を調査
    3.国内の研究状況調査:
    (1)昆虫ワークショツプ06年9/13-15(金沢)協賛;片岡、上田、小嶋が参加
    (2)名古屋大学理学部06年8/22-23;片岡が視察
    (3)昆虫科学シンポジウム(東京)06年11/18;片岡、嶋田が参加
    (4)農業生物資源研究所07年5/25;片岡が視察
    (5)神戸大学理学部、岡山大学理学部、山口大学理学部07年7/27-31;片岡が視察
    4.尾崎まみこ(神戸大学・理学部)を領域代表者とする新学術領域研究「仮;昆虫型環境適応プラン」を申請することになった。

  • Study on the molecular basis that underlies the dance language and aggressive behaviors of the honeybee.

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research

    Project Year :

    2004
    -
    2005
     

    KUBO Takeo

    　View Summary

    Aiming at analyzing the molecular basis of the honeybee social behaviors, particularly the dance communication and aggressive behaviors, identification and analysis of genes that are expressed in brain region- and/or behavior-selective manners were performed. The summary of the achievements is as follows :
    1.Brain regions associated with the dance communication were mapped by use of a novel immediate early gene, kakusei. The neural activity in the small-type Kenyon cells in the mushroom bodies (MBs) was enhanced in the dancer and forager honeybees.
    2.Genes whose expression is enhanced in the worker brain compared with the queen brain were screened. The HR38 homologue, which is involved in ecdysteriod-signaling, was identified as expressed preferentially in the small-type Kenyon cells in the forager brain.
    3.The expressions of two ecdysteroid-regulated genes, BR-C and E74, were concentrated in the MBs in the worker brain. In the MBs, they are expressed in Kenyon cell-selective manners, suggesting different roles of the Kenyon cell subtypes.
    4.The function of MBR-1, which is the C.elegans orthologue of Mblk-1, a MB-selective honeybee transcription factor, was analyzed genetically. The results revealed that the UNC-86-MBR-1 transcriptional cascade is involved in the pruning of excessive neuritis during development.
    5.A protocol for visual-probosis extension reflex associative learning was established using harnessed honeybees for the analysis of the mechanisms of visual learning. The results indicated that the chromatic adaptation also occurs in the honeybee.
    6.An epidemiologic study was performed for Kakugo virus (KV), which was originally identified from aggressive workers. KV was detected not only from attackers but also from the other worker populations in colonies that are heavily infected by KV.
    In addition, mites are suggested to mediate KV prevalence.
    These findings will provide important clues for further analysis of the molecular basis of the honeybee social behaviors.

  • 行動研究のモデル生物としてのミツバチの開発と利用

    日本学術振興会  科学研究費助成事業

    Project Year :

    2004
    　
    　
     

    久保 健雄, 佐々木 正己, 吉田 忠晴, 水波 誠, 蟻川 謙太郎, 松本 忠夫

    　View Summary

    本研究は、ミツバチが示す多彩な社会性行動やダンスコミュニケーションなどの高次行動を解析する「分子社会生物学」や「社会遺伝学」、「社会生理学」の分野における研究活動を、従来から発展してきた昆虫に関する神経生理学、行動生態学。神経行動学などの分野における研究活動と結びつけ、ミツバチを行動生物学の新しいモデル生物として開発・利用するそのための場の提供を目的とした。具体的には主に以下の2つの活動を行なった。
    (1)平成16年10月30日(土)に、東京大学大学院理学系研究科生物科学専攻(理学部2号館大講堂)において、全国の国立大学法人、公立大学、市立大学、国立研究機関、公立研究機関、民間企業の約45名の関連分野の研究者に呼び掛けて「社会性昆虫コンソーシアム」の設立会議を開催した。主たる参加者(30名)には、それぞれのグループの研究紹介ののち、その研究が今後のコンソーシアムの発展にどのように寄与しうるか説明していただき、研究交流を進めた。さらに、今後メーリングリストの準備とともにHP(=社会性昆虫コンソーシアムの媒体)を開設し、関連分野の情報公開・情報交換、共同研究の場を提供することを提案し、参加者全員から了解をいただくとともに、HP管理者(東京大学大学院理学系研究科助手 竹内秀明氏)を選出した。
    (2)「社会性昆虫コンソーシアム」のHP(http://www.honeybee.umin.jp/index.html)を開設し、メーリングリストを1月より稼働した。メーリングリストは既に会員により利用されている。HPは現在は仮設のものを公開しているが、管理を自動化したものを平成17年4月1日から稼働させる予定である。研究者紹介の他、関連分野をめぐる活動の情報公開、人事の案内、関連学会・雑誌へのリンク、ミツバチゲノム計画情報とのリンク等のサービスを行なう。将来的にはこのコンソーシアムの全体、あるいはその一部を母体にした共同研究や、特定領域研究の提案を視野に入れた活動を進めている。

  • 攻撃的なミツバチの脳に感染するRNAウイルスの感染ルートと攻撃行動への関与の証明

    日本学術振興会  科学研究費助成事業

    Project Year :

    2003
    -
    2004
     

    久保 健雄

    　View Summary

    ミツバチは社会性昆虫であり、そのコロニーでは個体間分業が見られる。本研究では、ミツバチの社会行動に関わる遺伝子候補として、2種類のRNA関連遺伝子:(1)攻撃蜂の脳から同定された新規なRNAウイルスと、(2)ミツバチ脳から同定された新規な非翻訳性核RNAに関する研究を進めた。
    (1)我々は既に、攻撃性が高い働き蜂の脳から新規な昆虫ピコルナ様ウイルス(Kakugoウイルス)を同定しており、働き蜂の攻撃行動がウイルス感染と関連する可能性を指摘してきた。今年度は、Kakugoウイルス感染がどの程度、働き蜂の攻撃行動と関連するか調べるため、疫学調査を行った。その結果、コロニーレベルでの感染量が低い条件ではKakugoウイルスは攻撃蜂に選択的に検出されたが、感染量が高いコロニーでは攻撃蜂以外の働き蜂(育児蜂や採餌蜂)にも感染が蔓延している例が見出された。これはKakugoウイルスの感染が限定された条件下でのみ攻撃性と関連することを示唆している。感染量が多いコロニーで、コロニー全体としての攻撃性が上昇しているかの検証が今後の重要な課題である。
    (2)我々は既にミツバチ脳で機能する非翻訳性核RNAの候補としてAncR-1を同定している。AncR-1は組織特異的に発現し、通常のmRNAと同様、スプライシングやpoly(A)付加を受ける一方で、in situハイブリダイゼーション法で核に局在して検出される点でmRNAとは異なる。今年度は、ミツバチ脳の核RNAを用いて作製したAncR-1 cDNAの構造を解析した結果、やはりAncR-1は核に濃縮されて存在し、プロセシングを受けることが確認された。これは組織特異的に発現するmRNA型の非翻訳性核RNAを同定した最初の例であり、ミツバチの行動制御の分子メカニズムだけでなく、新規なRNA分子種の発見という観点からも興味深い知見である。

  • ミツバチの脳に感染し、攻撃行動を誘発する新種のピコルナウイルスに関する研究

    日本学術振興会  科学研究費助成事業

    Project Year :

    2003
    　
    　
     

    久保 健雄

    　View Summary

    ミツバチは社会性昆虫であり、女王蜂(生殖カースト)が専ら産卵し、働き蜂(労働カースト)は育児や門番、採餌などの労働に携わる。ミツバチの働き蜂の天敵への攻撃行動は、自身が死亡する可能性が高い点で利他行動の代表例である。本研究では、働き蜂の攻撃行動を規定する分子的基盤を解析する目的で、攻撃的な働き蜂の脳で選択的に発現する遺伝子を検索した結果、攻撃蜂の脳に選択的に感染している新規なRNAウイルスを同定した。
    最初に、囮であるオオスズメバチに攻撃を仕掛けた働き蜂(攻撃蜂)と、スズメバチから逃避した働き蜂(逃避蜂)を採集し、それぞれの脳からRNAを抽出した。Differential display法により、攻撃蜂の脳に選択的に存在するRNAを検索した結果、kakugoと命名したRNAを同定した。cDNAクローニングの結果、kakugo RNAは様々なピコルナ様ウイルスのポリプロテインと有意な相同性を示すタンパク質をコードすることが判明した。このことは、kakugo RNAが新規なウイルスのゲノムRNAであり、攻撃蜂の脳はこのウイルスに感染していることを示唆している。また、攻撃蜂の細胞粗抽出液をショ糖密度勾配遠心により分離したところ、kakugo RNAは遊離のmRNAの他、ウイルス粒子に相当する分画にも検出された。さらに、攻撃蜂の細胞粗抽出液を非感染の働き蜂に注射したところ、3日後の脳でkakugo RNAが増幅していることが判明した。このことは、kakugo RNAが確かに感染性のあるウイルス粒子を形成することを示唆している。さらに、kakugoウイルスは攻撃蜂の脳では検出される一方で、育児蜂や採餌蜂では検出されず、攻撃行動とkakugoウイルスの感染が密接に関連することが示された。本研究は、動物の本能的な攻撃行動とウイルス感染が関連することを最初に示唆した点で学術的に価値が高い。

  • アフリカツメガエルの尾の再生に働く遺伝子の系統的検索と解析

    日本学術振興会  科学研究費助成事業

    Project Year :

    2001
    　
    　
     

    久保 健雄

    　View Summary

    研究代表者らは、脊椎動物の器官再生の分子機構を解明する目的で、器官再生能が高い両生類(アフリカツメガエル)において、幼生の尾の再生芽で特異的に発現する遺伝子をdifferential display法により検索し、neuronal pentraxin I(NP1)とコラーゲン遺伝子を同定している。
    本研究では、再生の初期に働く分子を調べる目的で、ツメガエル幼生の尾の切断後、再生芽形成に先立って発現する遺伝子を、differential display法で検索するとともに、NP1の発現をin situハイブリダイゼーション法で詳細に検討した。その結果、アポトーシス促進活性をもつHsp結合タンパク質の遺伝子が、再生芽の形成に先立って一過的に発現することを見出し、その完全長cDNAを得た。このタンパク質は、再生芽形成のための間隙の形成に働く可能性がある。またNP1遺伝子は尾の再生芽の間充織細胞に特異的に発現し、上皮細胞には発現しないことから、再生芽における間充織-上皮相互作用に関わると推測された。
    一方、同じく高い器官再生能をもつ昆虫(ワモンゴキブリ)では、Ca^<2+>依存性の体液性レクチンのファミリーが存在し、レクチン遺伝子が肢の再生芽に特異的に発現することが示されている。本研究では、ツメガエルの器官再生に働く分子の候補としてこのレクチンのツメガエルホモログの同定も試みた。解析の結果、ツメガエルにもCa^<2+>依存性の血清レクチンのファミリーが存在し、そのメンバーの遺伝子がやはり再生芽選択的に発現することが判明した。ただし、両者は構造上の有意な類似性はなく、機能的なホモログと考えられた。
    以上、本研究ではツメガエルの器官再生時に選択的に発現する遺伝子群を新しく同定し、その特徴的な発現様式を明らかにした。前段と後段の内容は各々、投稿準備中、投稿中となっている。

  • 消毒剤耐性変異の獲得に伴うMRSAのβラクタム系抗生物質耐性の上昇

    日本学術振興会  科学研究費助成事業

    Project Year :

    2000
    -
    2001
     

    関水 和久, 久保 健雄

    　View Summary

    本研究の目的は、消毒剤耐性を指標に分離されたオキサシリン強度耐性株から、消毒剤耐性かつオキサシリン耐性を導く遺伝子変畢を同定することである。この目的を違成するため、我々は、まず黄色ブドウ球菌の変異株から変異遺伝子を同定する系の構築を試みた。具体的には、増殖が温度感受性となる黄色プドウ球菌変異株を分離し、温度感受性を相補する遺伝子の同定を行える系の構築を行っ本研究の目的は、消毒剤耐性を指標に分離されたオキサシリン強度耐性株から、消毒剤耐性かつオキサシリン耐性を導く遺伝子変畢を同定することである。この目的を違成するため、我々は、まず黄色ブドウ球菌の変異株から変異遺伝子を同定する系の構築を試みた。具体的には、増殖が温度感受性となる黄色プドウ球菌変異株を分離し、温度感受性を相補する遺伝子の同定を行える系の構築を行った。
    まず、黄色ブドウ球菌を変異源EMMで処理した後、細胞増殖が温度感受性を示す変異株を分離した。次に、黄色ブドウ球菌のゲノムDNAのライブラリーを構築し、温度感受性黄色ブドウ球菌侍の温度感受性を相補する遺伝子断片の収得を行った。その結果、複数の独立した温度感受性株について、温度感受性を相補する遺伝子断片を得ることが出来た。温度感受性を相補する遺伝子断片の塩基配列決定を行い、さらに温度感受性を相補する断片の最少領域を決定する実験から、温度感受性の責任遺伝子としてdnaE, polC, dnaC遺伝子をそれぞれ同定した。dnaE, polC,遺伝子は共にDNAポリメレースをコードする。dnaE・PolCの温度感受性株を同一の菌から分離できたのは本研究が初めてである。この繕果は、最近のDNA複製において、2穫類のDNAポリメレースが役割分担を行っていることを初めて示唆した結果である。また、黄色ブドウ球菌より、dnaC遺伝子の温度感受性変異を分離したのも本研究が初めてである。

  • Development of antibiotics by monitoring the inhibition of the ATP-binding to DnaA, the initiator protein of chromosomal DNA replication in bacteria

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research

    Project Year :

    2000
    -
    2001
     

    SEKIMIZU Kazuhisa

    　View Summary

    Staphylococcus aureus causes opportunistic diseases for humans. In this study, we examined biochemical nature of Staphylococcus aureus DnaA protein, the replication initiator. Previous studies with Escherichia coli DnaA protein demonstrated that the ATP-binding form of DnaA protein is active, whereas the ADP-binding form is inactive. Purified Staphylococcus aureus DnaA proteins also showed high affinity for ATP and ADP. We showed that phosphatidylglycerol, an acidic phospholipids, stimulated the release of ADP from the ADP-DnaA complex, resulting in the activation of DnaA protein. Lysylphosphatidylglycerol, a basic phospholipid, was shown to inhibit the action of phosphatidylglycerol. Thus, phosphatidylglycerol may negatively regulate re-activation of DnaA protein. We propose here a new model of the regulation of initiation of DNA replication by increase in the content of basic phospholipids in cytoplasmic membranes.
    We isolated mutants of Staphylococcus aureus whose DNA replication were temperature-sensitive. By complementation analysis, we identified polC, dnaE, and dnaC genes which are essential for DNA replication in the bacteria. We also established an animal model of infectious diseases by using silkworms. The system will provide a useful screening system for medicines against infectious diseases.

  • Identification and analysis of genes involved in the intrinsic behaviors of the social insect (the honey bee)

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research

    Project Year :

    2000
    -
    2001
     

    KUBO Takeo

    　View Summary

    The honey bee is a social insect and the workers can communicate with other workers by dance language that contains information both on the direction and distance of the food source. In the honey bee brain, the mushroom bodies which are important for sensory integration, learning and memory in the insect brain, are characteristically well-developed and the intrinsic neuron (Kenyon cells) that constitute the mushroom bodies are classified into two subspecies : the large and small type-Kenyon cells. We previously identified two genes expressed preferentially in the large and small type-Kenyon cells (the Mblk-1 and the Ks-1 gene), as candidate genes involved in the highly advanced behavior of the honey bees. In this study, we isolated and characterized cDNAs for the Mblk-1 aad the Ks-1 genes.
    We found the Mblk-1 gene encodes a novel transcription factor and showed that this protein actually has transactivation activity. This is the first instance of a transcription factor expressed preferentially in the mushroom bodies of the insect brain and would be an important cue to understand the molecular basis of the insect brain function. The Mblk-1 homologue was also found in nematoda and mouse, suggesting that the function of Mblk-1 is conserved among various animals. On the contrary, the cDNA for the Ks-1 gene expressed preferentially in the small type-Kenyon cells encoded no significant ORF and the Ks-1 transcripts are located punctate in the nuclei of the small type-Kenyon cells. These results suggest that Ks-1 transcript functions as a non-translatable nuclear RNA to regulate the honey bee neuronal functions. This is the first instance of a neural cell type-preferential nuclear RNA and would be an important cue to analyze the function of a novel class RNA.
    In addition, the Q7 and QM1 genes expressed preferentially in the brain of the queens and the Agg-1 and Agg-2 genes expressed preferentially in the brain of the aggressive bees have been identified in this study, as candidate genes to be involved in the highly advanced behavior of the honey bees.

  • ミツバチの脳で領野特異的、行動特異的に発現する遺伝子の同定と解析

    日本学術振興会  科学研究費助成事業

    Project Year :

    2000
    　
    　
     

    久保 健雄

    　View Summary

    ミツバチは社会性昆虫であり、雌が女王蜂(生殖カースト)と働き蜂(労働カースト)に分化している。また、働き蜂は羽化後の日齢にともなって、育児から門番、採餌へと行動が変化(分業)する。ミツバチの脳では、感覚統合や記憶・学習の中枢であるキノコ体が顕著に発達しており、キノコ体を構成する介在神経細胞(ケニヨン細胞)の数はカースト間で異なること、またキノコ体の形態は育児蜂と採餌蜂で異なることが知られている。本研究では、ミツバチの社会行動を規定する脳内の分子的基盤を解明する目的で、ミツバチの行動依存的にキノコ体で発現する遺伝子をdifferential display法にて検索、解析した。
    その結果、働き蜂に比べて女王蜂のキノコ体で強く発現する遺伝子Q7を初めて同定した。この遺伝子の発現部域を解析する目的で、女王蜂と働き蜂の頭部と胸部、腹部での発現を調べたところ、女王蜂のキノコ体に加えて、女王蜂の腹部(卵巣)でも強く発現することが判明した。これらの知見は、Q7遺伝子産物が、産卵などの女王蜂固有な行動(脳)と、それに対応した生理状態(卵巣の発達)の協調的な制御に関わることを示唆しており、ミツバチのカースト分化の分子機構を知る上で重要な手掛かりを与えるものと思われる。現在、Q7遺伝子のcDNAクローニングを進めており、約10kbpに及ぶ配列を決定したが、これまでのところORFは見出されていない。この他にも本研究では、スズメバチへの攻撃性を指標に門番蜂を採取し、逃避蜂に比べて門番蜂の脳で選択的に発現する遺伝子の候補をdifferential display法で多数、同定した。さらに、キノコ体を構成する2種類の介在神経細胞(大型と小型のケニヨン細胞)のそれぞれに特異的に発現する遺伝子のcDNAクローニングを行った。

  • Molecular Mechanism of Insect Metamorphosis and Diapause

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research

    Project Year :

    2000
    　
    　
     

    YAMASHITA Okitsugu, SUZUKI Akinori, KUBO Takeo, KATAOKA Hiroshi, NATORI Syunji, NAGASAWA Hiromichi

    　View Summary

    The research project "Molecular Mechanism of Insect Metamorphosis and Diapause" (abbreviated as metamorphosis and diapause mechanism) was conducted for 4 years from 1996 to 1999 by 62 scientists in various fields. This project took aim at the molecular analysis of metamorphosis and diapause of insects that are the essential events supporting the prosperity of insects in this planet. Toward this goal research proposals employing molecular genetics, biochemical analysis, cellular biology and environmental physiology were encouraged, and large members of new genes and components responsible for synthesis of neuropeptide hormone, for its action, for non-self recognition of larval tissues, for social differentiation of honey bees and for environmental response. Part of research achievements was made public in an open symposium ; "University and Sciences" organized by the Ministry of Education, Science, Sports and Culture. Core members of this project have examined and summarized the results obtained by research members. Thc outline of the results has been presented to the authority for the final evaluation of research activity. Finally, the research activity of this project was collectively compiled to the booklet to distribute to a large number of scientists, specialists and offices in the related field.

  • Study of the regulatory mechanism of DnaA protein, the initiator of DNA replication in Escherichia coli.

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research

    Project Year :

    1999
    -
    2000
     

    SEKIMIZU Kazuhisa, KUBO Takeo

    　View Summary

    The purpose of this study is to understand the regulatory mechanism of DnaA protein, the initiator of DNA replication in Escherichia coli. The head investigator et al. previously reported that (i) DnaA protein has a high affinity for ATP, (ii) ATP bound to DnaA protein is hydrolyzed to form the ADP-binding form of DnaA protein, which is inactive for DNA replication, and, (iii) DNA polymerase III holoenzyme, the replicase in Escherichia coli, stimulates the hydrolysis of ATP bound to DnaA protein. In this study, we established a new method to determine adenine nucleotide bound to DnaA protein in vivo. Radio labeled cells with [^<32>P] orthophosphate were homogenized, and DnaA protein was recovered by immuno-precipitation, followed by analysis by thin layer chromatography. By using various temperature-sensitive mutants of DNA replication, we demonstrated that replication proteins responsible for the formation of the replication fork participate in the reaction of the hydrolysis ATP bound to the initiator protein. We also established a method to determine phospholipid bound to DnaA protein. The radiolabeled immuno-precipitates were extracted with organic solvents, and analyzed by thin layer chromatography. We detected cardiolipin and phosphatidylglycerol, which are previously shown to bind to DnaA protein in vitro, in the immuno-precipitates. The result demonstrates that DnaA protein binds to acidic phospholipids in cells.

  • Molecular mechanisms of self / non-self recognition and tissue-remodeling during metamorphosis.

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research

    Project Year :

    1996
    -
    1999
     

    KUBO Takeo, ASHIDA Masaaki, ISHIKAWA Hajime, NATORI Shunji, SAKURAI Sho, UENO Kohji

    　View Summary

    During metamorphosis of holometabolous insects, larval tissues that become unnecessary are eliminated by pupal hemocytes, whereas imaginal discs start to differentiate into adult tissues. In this research, we focused on molecules that drive insect metamorphosis and mainly analyzed molecular bases that underlie (1) recognition and elimimation of larval tissues and (2) formation of adult tissues during metamorphosis. We also analyzed (3) molecular basis of insect social behaviors as a new project, to gain more perspective in our research.
    Concerning project (1), we identified and cloned a novel hemocyte membrane protein that participates in recognition of larval tissues and a protease that is expressed in pupal mid gut of the flesh fly. We also determined complete genome sequence of Buchnera, a intracellular simbiont of aphid, to discuss possible evolution of intracellular simbiosis in insects. Concerning project (2), we have established an in vitro system of remodeling of larval central nervous system by adittion of ecdysone ; identified novel proteins (humoral lectins and extaracellular matrix proteins) that are expressed transiently during cockroach leg regeneration ; and identified and analyzed functions of novel genes that participate in morphogenesis in Drosophila (dad, brinker and Dally). Concerning project (3), we identified and analyzed genes expressed differently in the honeybee brain depending on their behaviors. We also analyzed functional flexibility of worker secretory (hypopharyngeal) gland cells depending on their division of labor. Many original and valuable findings have been obtained through this research and are expected to contribute to further development of this research area,

  • Identification and analysis of the genes that regurate the bahavior of the honeybee

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research

    Project Year :

    1997
    -
    1998
     

    KUBO Takeo

    　View Summary

    Honeybee is a social insect and many exquisite communications are performed among the colony members such as "dance launguage". However, the molecular basis that underlies such highly advanced types of honeybee behavior is largely unknown. The mushroom bodies (MB), which are supposed to be a main center of the senssory integration and memory, are characteristically well-developed in comparison with those of other insects. Thus, the increased complexity of the function and structure of the MB might be due to the highly advanced types of honeybee bahavior. Here, we searched for genes that are expressed preferentially in the MB by the differential display method as candidate genes that regulate the honeybee behavior, and obtained the following results.
    1. The intrinsic ceuron cells that consist the MB are subdivided into two classes : the large-and small-type Kenyon cells. A gene which is expressed specifically in the large-type Kenyon cells, termed M5, and a gene which is expressed specifically in the small-type Kenyon cells, termed Ks, are identified for the first time. cDNAs for these genes did not contain significant long open reading frames, suggesting that the M5 and Ks gene transcripts function as non-translated RNAs. In situ hybridization analysis of the M5 gene transcripts in the pupal brain revealed that the honeybee MB is a quadruple structure consisting of four clonal units, each of wich contains two distinct areas that express different genes. Furthure analysis of the functions of these gene products will reveal the subdivision of the inner structure of the MB that uinderlie the honeybee behavior.
    2. We showed that the expression of the genes for proteins involved in the intracellular calcium signal transduction was up-regulated in the MB of the honeybee. In contrast, in Drosophila, these genes were reported to be expressed ubiquitously thoroughout the brain cortex. Calcium signal transduction system of the neural cell has been reported to be involved in the estabrishment of memory and learning. Thus, it seems possible that the enhancement of the function of the calcium signaling contributes to the enhanced sinaptic plasticity of the MB of the honeybee. These findinG will be inportant from the perspective of comparative neurobiology of the insect.

  • Mechanisms of regeneration of insect imaginal discs.

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research

    Project Year :

    1995
    -
    1996
     

    KUBO Takeo, KOBAYASHI Ayako

    　View Summary

    1. Restoration of positional values of Sarcophaga imaginal discs on regeneration in vitro.
    When the 1/4 sectors of imaginal discs of Sarcophaga expressing wingles (wg) gene are removed and the remeining parts are cultured in the presence of 2.5 x 10^<-8>M 20-hydroxyecdyson, restoration of their morphology occurrs. Here, we isolated a cDNA for the Sarcophaga wg gene, and analyzed whether the lost positional values revealed by wg expression were restored when the remaining parts of imaginal discs were cultured in the presence of 20-hydroxyecdyson. As results, expression of the wg gene was detected in the regenrated areas, indicating that regeneration of imaginal discs in vitro minics in vivo regeneration in that it involves restoration of positional values.
    2.cDNA cloning and analysis of expression of the gene for lectins that localize in the regenerating legs of American cockroach.
    Two humoral lectins, termed 26-kDa lectin and regenectin, appear transiently in the regenerating legs of American cockroach. Among them, 26-kDa lectin localizes around epidermis of the regenerating legs in the middle stage of regeneratin, whereas regenectin localizes around regenetating muscle cells in the late stage. In this study, we isoalted cDNAs for these lectins and determined their primary structures. As results, these lectins were found to be C-type (Ca^<2+>-dependent) lectins and shares about 50% sequence identities with eact other. these results suggest that these lectins belong to the same protein family and play different roles in leg regeneration. The gene for regenectin was found to express transiently in the regenerating epidermis, suggesting that it functions in the formation of muscle cells in a paracrine manner by synthesized and secreted by the epidermis.
    This is the first instance that identified molecules (20-hydroxyecdyson and humoral lectins) that is involved in the regeneration in insects and woud be an important clue to better undrstand the molecular mechanisms of organ regeneration.

  • 昆虫の変態・休眠の分子機構に関する研究の企画調査

    日本学術振興会  科学研究費助成事業

    Project Year :

    1995
    　
    　
     

    山下 興亜, 鈴木 昭憲, 相薗 泰生, 桜井 勝, 久保 健雄, 片岡 宏誌

    　View Summary

    1.重点課題と研究組織の確立
    平成8年度から重点領域研究として「昆虫の変態・休眠の分子機構」を発足ために,わが国における研究動向,研究実績,研究組織,国際比較を全体会議を開いて検討した.その結果,研究課題としては環境-生体情報-情報伝達系の体系的な解明と自己認識機構の発育変動を中心とした形態形成と器官再生による変態理解を取り上げること,若手研究者の創意を最大限発揮する研究組織を組むこと,研究成果の評価を多面的に行い新規分野の開拓につなげること,を確認した.
    2.研究推進の方策
    専門分野の代表者により,研究方法の改良・開拓,対象昆虫種の選定・拡大,大型機器の共同利用,日常的な情報交換の円滑化などを検討し,各研究者の独自性をもとにした協力・共同研究を推進し,総体として活発な研究活動を保証する方策を作成した.

  • Defense mechanism of insect (Sarcophaga peregrina)

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research

    Project Year :

    1992
    -
    1994
     

    NATORI Shunji, KURATA S, KUBO T

    　View Summary

    Insect have developed a unique defense system against various outside stimuli such as microbial infection and integmental injury. We have been studying various defense proteins of Sarcophaga peregrina (the flesh fly). These proteins include a lectin and several antibacterial proteins. Throughout these studies, we came to the conclusion that there are two important novel aspects of these defense proteins. One is that a single defense protein probably plays at least two roles in defense and development. Insects probably have the flexibility to use one defense protein in multiple ways. The other interesting aspect is the phylogenetical significance of the insect defense system. Insect defense system is possibly a prototype of the mammalian defense system. Except these two important aspects, we obtained various important findings on insect immnity. Most of these findings have been published.

  • ワモンゴキブリの体液性レクチンの肢の再生に於ける機能解析と構造決定

    日本学術振興会  科学研究費助成事業

    Project Year :

    1991
    -
    1992
     

    久保 健雄

    　View Summary

    研究代表者らはこれまでに、ワモンゴキブリの体液から共通抗原性を示す3種類のC型レクチンを精製し、その内regenectin、26kDa lectinと命名したレクチンが肢の再生芽に一過的に出現することを見いだしている。Regenectinが再生芽の筋細胞の周囲に局在するのに対し、26kDa lectinは上皮組織の外側に局在する。従って、ワモンゴキブリの肢の再生芽形成過程では、これらのレクチンが異なる組織の細胞接着分子として働くことにより、再生芽の空間的な秩序の形成に働くものと推察される。更に、regenectinは正常な発生過程には発現しないことから、再生筋細胞の接着に特異的に働く分子であると考えられる。
    今年度は、regenectinの肢の筋細胞における局在を、組織切片の染色と蛍光抗体法を組み合わせることにより詳細に解析した結果、このレクチンは筋細胞の基底膜か細胞膜の部分と、筋細胞に接する上皮細胞の内側の表面にも局在することを見いだした。このことから、regenectinは再生芽の筋細胞同士の接着に働くばかりでなく、上皮組織と筋組織という異なる組織の細胞の選別にも関与する可能性を新たに指摘できた。更に、regenectinと26kDa lectinの部分アミノ酸配列を決定した結果、これらのレクチンの一次構造は、既に我々の研究室で同じワモンゴキブリの成虫体液から精製し、cDNAクロ-ニングによって一次構造が明らかとなっているLPS結合蛋白(LPSの糖鎖部分を認識するC型レクチンの一種)の一次構造と高い類似性を示すことを見いだした。従って、これらのレクチンは、実際に一次構造の類似したファミリ-を構成していることが示された。次年度は、これらのレクチンの再生における役割を解析するとともに、cDNAクロ-ニングによるレクチンの構造解析を行い、再生芽におけるレクチンmRNAの発現を解析することを目的として研究を推進する。

▼display all

Misc

  • Developmental stage-specific expression analysis of Mblk-1, a transcription factor involved in ecdysteroid-signaling in the honey bee brain

    松村泰宏, 河野大輝, 久保健雄

    昆虫と自然   56 ( 1 )  2021

    J-GLOBAL

  • ミツバチの社会性行動の分子生物学 遺伝子から探るミツバチの脳の仕組み (特集 ミツバチ新時代)

    河野 大輝, 久保 健雄

    遺伝 : 生物の科学   73 ( 6 ) 596 - 602  2019.11

    CiNii

  • Correction: Gene expression and immunohistochemical analyses of mKast suggest its late pupal and adult-specific functions in the honeybee brain (vol 12, e0183522, 2017)

    Atsuhiro Yamane, Hiroki Kohno, Tsubomi Ikeda, Kumi Kaneko, Atsushi Ugajin, Toshiyuki Fujita, Takekazu Kunieda, Takeo Kubo

    PLOS ONE   12 ( 10 )  2017.10

    Other  

    DOI

  • Development of genome-editing techniques in the honeybee : its future prospects

    河野 大輝, 佐々木 哲彦, 久保 健雄

    昆虫と自然   52 ( 10 ) 36 - 39  2017.09

    CiNii

  • 放射線耐性動物クマムシの新規タンパク質によるヒト培養細胞におけるDNA傷害の抑制と放射線耐性の向上

    橋本拓磨, 斎藤裕樹, 秦裕子, 榎本敦, 堀川大樹, 堀川大樹, 荒川和晴, 片山俊明, 豊田敦, 尾山大明, 宮川清, 久保健雄, 國枝武和

    日本放射線影響学会大会抄録(Web)   59th  2016

    J-GLOBAL

  • 【昆虫の生物機能の解明と創薬・医療への応用】 ミツバチの「尻振りダンス」の分子・神経的基盤の解析

    金子 九美, 久保 健雄

    生化学   86 ( 5 ) 588 - 594  2014.10

  • 放射線耐性を示すクマムシの新規タンパク質S261によるDNA損傷の抑制

    橋本拓磨, 斎藤裕樹, 尾山大明, 秦裕子, 榎本敦, 宮川清, 桑原宏和, 堀川大樹, 豊田敦, 片山俊明, 荒川和晴, 藤山秋佐夫, 久保健雄, 國枝武和

    日本放射線影響学会大会講演要旨集   57th   121  2014.09

    J-GLOBAL

  • 放射線耐性を示すクマムシの新規タンパク質によるDNA損傷の抑制

    橋本拓磨, 斎藤裕樹, 尾山大明, 秦裕子, 榎本敦, 宮川清, 桑原宏和, 堀川大樹, 豊田敦, 藤山秋佐夫, 久保健雄, 國枝武和

    日本動物学会大会予稿集   85th   158  2014.08

    J-GLOBAL

  • アフリカツメガエル幼生尾の再生芽増殖細胞に選択的に発現する遺伝子の網羅的検索と解析

    辻岡洋, 國枝武和, 加藤由起, 白髭克彦, 久保健雄

    日本動物学会大会予稿集   85th   94  2014.08

    J-GLOBAL

  • アフリカツメガエル幼生尾の再生芽増殖細胞に選択的に発現する遺伝子の網羅的検索と解析

    辻岡洋, 國枝武和, 加藤由起, 白髭克彦, 久保健雄

    日本細胞生物学会大会要旨集   66th   125  2014.05

    J-GLOBAL

  • 放射線耐性を示すクマムシの新規タンパク質S261によるDNA損傷の抑制

    橋本拓磨, 斎藤裕樹, 尾山大明, 秦裕子, 榎本敦, 宮川清, 桑原宏和, 堀川大樹, 豊田敦, 片山俊明, 荒川和晴, 藤山秋佐夫, 久保健雄, 國枝武和

    日本分子生物学会年会プログラム・要旨集(Web)   37th   3P-0001 (WEB ONLY)  2014

    J-GLOBAL

  • アフリカツメガエル幼生尾の再生芽増殖細胞に選択的に発現する遺伝子の解析

    辻岡洋, 國枝武和, 加藤由起, 白髭克彦, 久保健雄

    日本分子生物学会年会プログラム・要旨集(Web)   37th   1P-0574 (WEB ONLY)  2014

    J-GLOBAL

  • ツメガエル幼生尾の再生芽増殖細胞のRNA‐seq解析

    辻岡洋, 加藤由起, 國枝武和, 白髭克彦, 久保健雄

    日本動物学会大会予稿集   84th   115  2013.08

    J-GLOBAL

  • セイヨウミツバチApis mellifera L.脳でキノコ体に高度に選択的に発現する3遺伝子の同定

    末次翔太, 竹内秀明, 國枝武和, 久保健雄

    日本動物学会大会予稿集   84th   158  2013.08

    J-GLOBAL

  • メダカの配偶者選択の分子神経基盤

    奥山輝大, 横井佐織, 阿部秀樹, 磯江泰子, 末廣勇司, 今田はるか, 島田敦子, 川崎隆史, 弓場俊輔, 谷口善仁, 亀井保博, 田中実, 成瀬清, 武田洋幸, 岡良隆, 久保健雄, 竹内秀明

    日本動物学会大会予稿集   84th   137  2013.08

    J-GLOBAL

  • アフリカツメガエル幼生の尾の再生芽増殖細胞の単離と遺伝子発現プロファイリング

    辻岡洋, 加藤由起, 國枝武和, 白髭克彦, 久保健雄

    日本分子生物学会年会プログラム・要旨集(Web)   36th   2P-0635 (WEB ONLY)  2013

    J-GLOBAL

  • セイヨウミツバチのローヤルゼリーとその由来外分泌腺のプロテオミクスを用いた解析

    藤田俊之, 近藤裕子, 秦裕子, 國枝武和, 尾山大明, 久保健雄

    日本生化学会大会(Web)   85th   2P-661 (WEB ONLY)  2012

    J-GLOBAL

  • 免疫抑制による"再生不応期"のアフリカツメガエル幼生尾の再生能の人為的活性化 脊椎動物の器官再生能の有無を決定する要因を探る

    深澤 太郎, 直良 悠子, 國枝 武和, 久保 健雄, 理研バイオリソースセンター生体応答情報技術開発サブチーム

    細胞工学   30 ( 4 ) 418 - 423  2011.03

    CiNii

  • ニホンミツバチ脳における熱殺蜂球形成行動時の興奮領域の解析

    宇賀神篤, 木矢剛智, 國枝武和, 小野正人, 吉田忠晴, 久保健雄

    生化学     ROMBUNNO.1P-0843  2010

    J-GLOBAL

  • 視覚依存の配偶者選別において行動異常を示すメダカ変異体の同定

    奥山輝大, 末廣勇司, 今田はるか, 阿部秀樹, 島田敦子, 田中実, 成瀬清, 武田洋幸, 岡良隆, 久保健雄, 竹内秀明

    生化学     ROMBUNNO.1P-0845  2010

    J-GLOBAL

  • Analysis of the brain substructures using region-preferential genes of the honeybee

       2009.12

    Research paper, summary (national, other academic conference)  

  • Analysis of Schooling-like Behavior of Medaka Fish and Identification of a Telencephalon-and-Hypothalamus-Preferential Gene in the Medaka Brain

    Haruka Imada, Yuji Suehiro, Teruhiro Okuyama, Daisuke Kurabayashi, Atsuko Shimada, Kiyoshi Naruse, Hiroyuki Takeda, Takeo Kubo, Hideaki Takeuchi

    Proceeding of the Third International Symposium on Mobiligence （査読有り国際紀要）     295 - 298  2009.11  [Refereed]

    Meeting report  

  • Analysis of neural mechanism for the optomotor response of the medaka fish

    Yuji Suehiro, Teruhiro Okuyama, Haruka Imada, Kiyoshi Naruse, Daisuke Kurabayashi, Atsuko Shimada, Hiroyuki Takeda, Takeo Kubo, Hideaki Takeuchi

    Proceeding of the Third International Symposium on Mobiligence （査読有り国際紀要）     308 - 311  2009.11  [Refereed]

    Meeting report  

  • 不応期アフリカツメガエル幼生尾の再生誘導に充分な免疫抑制剤FK506の処理時期の決定

    菱田祐子, 深澤太郎, 國枝武和, 久保健雄

    日本動物学会大会予稿集   79th   125  2008.08

    J-GLOBAL

  • 昆虫の生存戦略の分子機構 攻撃的なミツバチから見つかったKakugoウイルスの疫学的解析 ミツバチに見るウイルス-宿主間相互作用

    藤 幸知子, 久保 健雄

    化学と生物   46 ( 7 ) 491 - 495  2008.07

    DOI CiNii

  • Identification and analysis of genes that are expressed strongly in the optic lobes in the honeybee brain

    K. Kaneko, S. Hori, H. Takeuchi, R. K. Paul, T. Fujiyuki, K. Shirai, A. Wakamoto, T. Kubo

    FEBS JOURNAL   275   147 - 147  2008.06

    Research paper, summary (international conference)  

  • Differential expression of hormone receptor-like in 38 (HR38) in the honeybee brain depending on the caste differentiation and division of labor of the workers

    Y. Yamazaki, K. Shirai, R. K. Paul, T. Fujiyuki, A. Wakamoto, H. Takeuchi, T. Kubo

    FEBS JOURNAL   275   144 - 144  2008.06

    Research paper, summary (international conference)  

  • Mushroom body-selective expression of genes for proteins involved in juvenile hormone metabolism and calcium-signaling in honeybee brain

    Y. Uno, T. Fujiyuki, M. Morioka, T. Kubo

    FEBS JOURNAL   275   151 - 151  2008.06

    Research paper, summary (international conference)  

  • 小型魚類の適応的行動(視運動反応, 群れ行動)の脳情報処理原理とその神経・分子的基盤の解明を目指して

    竹内 秀明, 末廣 勇司, 今田 はるか, 奥山 輝大, 久保 健雄

    計測と制御 = Journal of the Society of Instrument and Control Engineers   46 ( 12 ) 922 - 927  2008.01

    Book review, literature introduction, etc.  

    　View Summary

    研究紹介に日本語解説

    DOI CiNii

  • 細胞外分泌型アデノシンデアミナーゼの脊椎動物における生理機能の解析

    神垣ひろこ, 小野塚潤, 飯島亮子, 山口真二, 國枝武和, 久保健雄, 名取俊二, 本間光一

    日本薬学会年会要旨集   127th ( 2 ) 105  2007.03

    J-GLOBAL

  • 社会性昆虫ミツバチのゲノムが語るものーミツバチのゲノムから社会性の理解へー

    國枝武和, 藤幸知子, 久保健雄

    生物の科学・遺伝   61   4 - 7  2007

  • Identification of mandibular gland proteins expressed in a caste-selecttve manner in the honeybee

    Makoto Hasegawa, Tomoko Fujiyuki, Mizue Morioka, Takeo Kubo

    ZOOLOGICAL SCIENCE   23 ( 12 ) 1209 - 1209  2006.12

    Research paper, summary (international conference)  

  • Identification and analysis of genes that are expressed strongly in the optic lobes in the honeybee brain

    Kumi Kaneko, Sayaka Hori, Hideaki Takeuchi, Rajib Paul, Tomoko Fujiyuki, Kenichi Shirai, Akiko Wakamoto, Takeo Kubo

    ZOOLOGICAL SCIENCE   23 ( 12 ) 1191 - 1191  2006.12

    Research paper, summary (international conference)  

  • 動物脳の高次中枢選択的に発現する新規転写因子Mblk-1/MBR-1のリン酸化による活性制御

    久保 健雄

    日本応用酵素協会誌   ( 40 ) 63 - 63  2006.02

  • Prevalence of Kakugo virus in the honeybee Apis mellifera L. population analyzed by quantitative reverse transcription-PCR

    Tomoko Fujiyuki, Hideaki Takeuchi, Seii Ohka, Masato Ono, Akio Nomoto, Takeo Kubo

    ZOOLOGICAL SCIENCE   22 ( 12 ) 1482 - 1482  2005.12

    Research paper, summary (international conference)  

  • 線虫 C.elegans をモデルとした神経突起除去の分子機構の解析

    HAYASHI YU, KAGE ERIKO, HIROTSU TAKAAKI, KUNITOMO HIROFUMI, TAKEUCHI HIDEAKI, IINO YUICHI, KUBO TAKEO

    日本分子生物学会年会講演要旨集   28th   294  2005.11

    J-GLOBAL

  • 線虫C.elegansの嗅覚可塑性に関わる転写因子MBR-1の軸索除去における機能解析(Analysis of the function of MBR-1,a C.elegans transcription factor involved in olfactory plasticity,in axon pruning)

    林 悠, 鹿毛 枝里子, 広津 崇亮, 國友 博文, 竹内 秀明, 飯野 雄一, 久保 健雄

    神経化学   43 ( 2-3 ) 526 - 526  2004.08

  • ミツバチをモデル生物とした分子行動生物学

    竹内秀明, 國枝武和, 徳廣(澤田, 美由紀, 朴正敏, 藤幸知子, 久保健雄

    蛋白質核酸酵素   49 ( 16 ) 2542 - 2548  2004

  • ミツバチの社会性をめぐる分子生物学——昆虫の社会性の進化にどのようにアプローチするか

    久保健雄, 大橋一晶, 澤田美由紀, 上川内あづさ, 竹内秀明

    蛋白質核酸酵素   45 ( 7 ) 1229 - 1236  2000.05

    PubMed CiNii

  • Identification and analysis of the gene which selectively appears to the mushroom body of the honeybee.

    TAKEUCHI Hideaki, KAMIKOUCHI Azusa, OHARA Maya, SAWATA Miyuki, YOSHINO Daisuke, KUBO Takeo

    Dobutsu seiri   16 ( 4 ) 266 - 277  1999.12

    DOI CiNii

  • Gene organization and chromosome mapping of the testis-specific S-II

    T Ito, MF Seldin, MM Taketo, T Kubo, S Natori

    MAMMALIAN GENOME   9 ( 11 ) 915 - 917  1998.11

    DOI PubMed

  • Molecular Sociobiology of the Honeybee

    KUBO Takeo, OHASHI Kazuaki, TAKEUCHI Hideaki, SAWATA Miyuki, KAMIKOUCHI Azusa, NATORI Shunji

    KAGAKU TO SEIBUTSU   34 ( 12 ) 793 - 798  1997.01

    DOI CiNii

  • Spermatocyte-specific expression of the gene for mouse testis-specific transcription elongation factor S-II

    T Ito, QH Xu, H Takeuchi, T Kubo, S Natori

    FEBS LETTERS   385 ( 1-2 ) 21 - 24  1996.04

    DOI PubMed CiNii

  • 化学療法下の乳癌患者血漿中で発見された未知な物質を2′-deoxycytidineと同定

    吉岡 正則, Abu-Zeid Medhat, 久保 健雄

    Biological & Pharmaceutical Bulletin   17 ( 2 ) 169 - 172  1994.02

  • ワモンゴキブリの肢の再生芽に局在する体液性レクチン再生芽の形態形成を制御する因子か

    久保 健雄, 名取 俊二

    化学と生物   28 ( 8 ) 489 - 491  1990.08

    CiNii

▼display all

 

Syllabus

▼display all

 

Research Institute

  • 2026
    　
    　

    Waseda Research Institute for Science and Engineering   Concurrent Researcher