Updated on 2022/04/20

写真a

 
KINOSHITA, Makoto
 
Organization
Graduate School of Science Professor
Graduate School
Graduate School of Science
Undergraduate School
School of Science
Title
Professor
External link

Degree 1

  1. Doctor (Medicine) ( 1995.3   Kyoto University ) 

Research Interests 14

  1. molecular and cellular biology

  2. molecular and cellular neuroscience

  3. 神経化学

  4. 電子顕微鏡

  5. 細胞骨格

  6. 細胞分裂

  7. 神経突起

  8. パーキンソン病

  9. ノックアウトマウス

  10. ドーパミン

  11. トランスポーター

  12. セプチン

  13. シナプス

  14. グリア細胞

Research Areas 6

  1. Others / Others  / Molecular Biology

  2. Others / Others  / Nerve Anatomy and Neuropathology

  3. Others / Others  / Animal Physiology/Behavior

  4. Life Science / Cell biology

  5. Life Science / Neuroscience-general

  6. Life Science / Experimental pathology

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Current Research Project and SDGs 1

  1. Molecular and physiological functions of the septin cytoskeletal system

Research History 9

  1. Nagoya University   Division of Biological Science   Professor

    2009.4

  2. Kyoto University   Lecturer

    2007.4 - 2009.3

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    Country:Japan

  3. JST   PRESTO   Researcher

    2003.10 - 2007.3

  4. Kyoto University   Biochemistry and Cell Biology Group, HMRO   Designated associate professor

    2003.4 - 2007.3

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    Country:Japan

  5. Harvard Medical School   Department of Cell Biology   Researcher

    2000.1 - 2003.6

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    Country:United States

  6. Kyoto University   Assistant

    1995.4 - 2001.3

  7. Cancer Institute (Japanese Foundation of Cancer Research)   Department of Viral Oncology

    1992.4 - 1995.3

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    Country:Japan

  8. (財) 田附興風会 医学研究所 北野病院   研修医   医師

    1990.4 - 1991.3

  9. Kyoto University

    1989.6 - 1990.3

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Education 2

  1. Kyoto University   Graduate School of Medicine

    1991.4 - 1994.3

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    Country: Japan

  2. Kyoto University   Faculty of Medicine

    1983.4 - 1989.3

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    Country: Japan

Professional Memberships 6

  1. Society for Neuroscience   Regular Member

  2. American Society for Cell Biology   Regular Member

  3. 日本分子生物学会   正会員

  4. 日本生化学会

  5. Japan Neuroscience Society   Regular Member

  6. The Japanese Society for Neurochemistry   Regular Member

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Committee Memberships 20

  1. 名古屋大学動物実験委員会   委員  

    2017.4   

  2. 次世代脳プロジェクト実行委員会   実行委員  

    2017.1 - 2019.3   

  3. 理学研究科附属ニューロサイエンス研究センター運営委員会   委員  

    2017.4   

  4. 国際プログラム群理系小部会   委員  

    2016.4 - 2020.3   

  5. 外国人留学生特別選抜委員会   委員  

    2016.4   

  6. 生命理学科将来計画委員会   委員  

    2016.4   

  7. G30教育生活連絡会議   委員  

    2017.4 - 2020.3   

  8. 理学研究科動物実験委員会   委員長  

    2014.4   

  9. 東山地区動物実験施設実務委員会   委員  

    2013.7   

  10. 臨海実験所協議会   協議員  

    2013.4 - 2019.3   

  11. 藤田保健衛生大学 脳関連遺伝子機能網羅的解析拠点 運営委員会   運営委員  

    2015.4   

  12. 理学研究科節電対策ワーキンググループ   委員  

    2014.4 - 2020.3   

  13. 文部科学省新学術領域研究 包括型脳科学研究推進支援ネットワーク   研究集会委員  

    2011.4 - 2016.3   

  14. 全学バイオセーフティー委員会   委員  

    2021.4   

  15. 全学人事プロセス委員会   学術委員  

    2020.4   

  16. 理学研究科建築委員会   委員  

    2019.4   

  17. 入学問題委員会   委員及び委員長(2019)  

    2017.4 - 2020.3   

  18. 全学安全保障委員会   委員  

    2014.4 - 2019.3   

  19. ベンチャー・ビジネス・ラボラトリー事業委員会   委員  

    2010.4 - 2012.3   

  20. 脳の医学・生物学研究会   幹事  

    2010   

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    Committee type:Academic society

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Papers 101

  1. Mice with cleavage-resistant N-cadherin exhibit synapse anomaly in the hippocampus and outperformance in spatial learning tasks Reviewed

    Asada-Utsugi M., Uemura K., Kubota M., Noda Y., Tashiro Y., Uemura T. M., Yamakado H., Urushitani M., Takahashi R., Hattori S., Miyakawa T., Ageta-Ishihara N., Kobayashi K., Kinoshita M., Kinoshita A.

      Vol. 14 ( 1 )   2021.1

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    Language:English   Publishing type:Research paper (scientific journal)  

    DOI: 10.1186/s13041-021-00738-1

    Web of Science

  2. Developmental and postdevelopmental roles of septins in the brain. Invited Reviewed International journal

    Natsumi Ageta-Ishihara, Makoto Kinoshita

    Neuroscience research     2020.11

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    Authorship:Last author, Corresponding author   Language:English   Publishing type:Research paper (scientific journal)  

    Morphogenetic processes during brain development and postdevelopmental remodeling of neural architecture depend on the exquisite interplay between the microtubule- and actin-based cytoskeletal systems. Accumulation of evidence indicates cooperative roles of another cytoskeletal system composed of the septin family. Here we overview experimental findings on mammalian septins and their hypothetical roles in the proliferation of neural progenitor cells, neurite development, synapse formation and regulations. The diverse, mostly unexpected phenotypes obtained from gain- and loss-of-function mutants point to unknown molecular network to be elucidated, which may underlie pathogenetic processes of infectious diseases and neuropsychiatric disorders in humans.

    DOI: 10.1016/j.neures.2020.08.006

    PubMed

  3. Chr21 protein-protein interactions: enrichment in products involved in intellectual disabilities, autism and Late Onset Alzheimer Disease International coauthorship

    Julia Viard, Yann Loe-Mie, Rachel Daudin, Malik Khelfaoui, Christine Plancon, Anne Boland, Francisco Tejedor, Richard L. Huganir, Eunjoon Kim, Makoto Kinoshita, Guofa Liu, Volker Haucke, Thomas Moncion, Eugene Yu, Valérie Hindie, Henri Bléhaut, Clotilde Mircher, Yann Herault, Jean-François Deleuze, Jean-Christophe Rain, Michel Simonneau, Aude-Marie Lepagnol-Bestel

    BioRxiv     2019.12

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    Language:English   Publishing type:Research paper (scientific journal)   Publisher:Cold Spring Harbor Laboratory  

    <title>ABSTRACT</title>Intellectual disability (ID) found in Down syndrome (DS), which is characterized by an extra copy of 234 genes on Chr21 is poorly understood. We first used two DS mouse models that either display an extra copy of the <italic>Dyrk1A</italic> gene or of the mouse Chr16 syntenic region. Exome sequencing of transcripts deregulated in embryonic hippocampus uncovers enrichment in genes involved in chromatin and synapse respectively. Using large-scale yeast two-hybrid screen (154 distinct screens) of human brain library containing at least 10<sup>7</sup> independent fragments, we identified 3,636 novel protein-protein interactions with an enrichment of direct interactors of both Chromosome 21(Hsa21) baits and rebounds in ID-related genes. Using proximity ligation assays, we identified that Hsa21-encoded proteins are located at the dendritic spine postsynaptic density in a protein network located at the dendritic spine post synapse. Hsa21 DYRK1A and DSCAM that confers a ~ 20-fold increase in Autism Spectrum Disorders (ASDs) are part of this dendritic spine postsynaptic network. We found that a DSCAM intracellular domain binds either DYRK1A or DLGs that are multimeric scaffolds for the clustering of receptors, ion channels, and associated signaling proteins. The DYRK1A-DSCAM interaction is conserved from drosophila to humans. The identified postsynaptic.network is enriched in ARC-related synaptic plasticity, ASDs and Late-Onset Alzheimer Disease. Altogether, these results emphasize links between DS and brain diseases with complex genetics.

    DOI: 10.1101/2019.12.11.872606

  4. PCP-dependent transcellular regulation of actomyosin oscillation facilitates convergent extension of vertebrate tissue Reviewed International coauthorship

    Asako Shindo, Yasuhiro Inoue, Makoto Kinoshita, John B. Wallingford

    Developmental Biology   Vol. 446 ( 2 ) page: 159 - 167   2019.2

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    Language:English   Publishing type:Research paper (scientific journal)   Publisher:Elsevier BV  

    DOI: 10.1016/j.ydbio.2018.12.017

    Web of Science

  5. UBL3 modification influences protein sorting to small extracellular vesicles Reviewed

    Hiroshi Ageta, Natsumi Ageta-Ishihara, Keisuke Hitachi, Ozge Karayel, Takanori Onouchi, Hisateru Yamaguchi, Tomoaki Kahyo, Ken Hatanaka, Koji Ikegami, Yusuke Yoshioka, Kenji Nakamura, Nobuyoshi Kosaka, Masashi Nakatani, Akiyoshi Uezumi, Tomihiko Ide, Yutaka Tsutsumi, Haruhiko Sugimura, Makoto Kinoshita, Takahiro Ochiya, Matthias Mann, Mitsutoshi Setou, Kunihiro Tsuchida

    Nature Communications   Vol. 9 ( 1 )   2018.12

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    Language:English   Publishing type:Research paper (scientific journal)   Publisher:Springer Science and Business Media LLC  

    DOI: 10.1038/s41467-018-06197-y

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    PubMed

    Other Link: http://www.nature.com/articles/s41467-018-06197-y

  6. CDC42EP4, a perisynaptic scaffold protein in Bergmann glia, is required for glutamatergic tripartite synapse configuration. Invited Reviewed International journal

    Natsumi Ageta-Ishihara, Kohtarou Konno, Maya Yamazaki, Manabu Abe, Kenji Sakimura, Masahiko Watanabe, Makoto Kinoshita

    Neurochemistry International   Vol. 119   page: 190 - 198   2018.10

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    Authorship:Last author, Corresponding author   Language:English   Publishing type:Research paper (scientific journal)   Publisher:Elsevier {BV}  

    Configuration of tripartite synapses, comprising the pre-, post-, and peri-synaptic components (axon terminal or bouton, dendritic spine, and astroglial terminal process), is a critical determinant of neurotransmitter kinetics and hence synaptic transmission. However, little is known about molecular basis for the regulation of tripartite synapse morphology. Previous studies showed that CDC42EP4, an effector protein of a cell morphogenesis regulator CDC42, is expressed exclusively in Bergmann glia in the cerebellar cortex, that it forms tight complex with the septin heterooligomer, and that it interacts indirectly with the glutamate transporter GLAST and MYH10/nonmuscle myosin ΙΙB. Scrutiny of Cdc42ep4-/- mice had revealed that the CDC42EP4-septins-GLAST interaction facilitates glutamate clearance, while the role for CDC42EP4-septins-MYH10 interaction has remained unsolved. Here, we find anomalous configuration of the tripartite synapses comprising the parallel fiber boutons, dendritic spines of Purkinje cells, and Bergmann glial processes in Cdc42ep4-/- mice. The complex anomalies include 1) recession of Bergmann glial membranes from the nearest active zones, and 2) extension of nonactive synaptic contact around active zone. In line with the recession of Bergmann glial membranes by the loss of CDC42EP4, overexpression of CDC42EP4 in heterologous cells promotes cell spreading and partitioning of MYH10 to insoluble (i.e., active) fraction. Paradoxically, however, Cdc42ep4-/- cerebellum contained significantly more MYH10 and N-cadherin, which is attributed to secondary neuronal response mainly in Purkinje cells. Given cooperative actions of N-cadherin and MYH10 for adhesion between neurons, we speculate that their augmentation may reflect the extension of nonactive synaptic contacts in Cdc42ep4-/- cerebellum. Transcellular mechanism that links the absence of CDC42EP4 in Bergmann glia to the augmentation of N-cadherin and MYH10 in neurons is currently unknown, but the phenotypic similarity to GLAST-null mice indicates involvement of the glutamate intolerance. Together, the unique phenotype of Cdc42ep4-/- mice provides a clue to novel molecular network underlying tripartite synapse configuration.

    DOI: 10.1016/j.neuint.2018.01.003

    Web of Science

    PubMed

  7. Septin-dependent remodeling of cortical microtubule drives cell reshaping during epithelial wound healing. Reviewed International coauthorship International journal

    Asako Shindo, Anastasia Audrey, Maki Takagishi, Masahide Takahashi, John B Wallingford, Makoto Kinoshita

    Journal of Cell Science   Vol. 131 ( 12 ) page: jcs212647 - jcs212647   2018.6

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    Authorship:Last author   Language:English   Publishing type:Research paper (scientific journal)   Publisher:The Company of Biologists  

    Wounds in embryos heal rapidly through contraction of the wound edges. Despite well-recognized significance of the actomyosin purse string for wound closure, roles for other cytoskeletal components are largely unknown. Here, we report that the septin cytoskeleton cooperates with actomyosin and microtubules to coordinate circumferential contraction of the wound margin and concentric elongation of wound-proximal cells in Xenopus laevis embryos. Microtubules reoriented radially, forming bundles along lateral cell cortices in elongating wound-proximal cells. Depletion of septin 7 (Sept7) slowed wound closure by attenuating the wound edge contraction and cell elongation. ROCK/Rho-kinase inhibitor-mediated suppression of actomyosin contractility enhanced the Sept7 phenotype, whereas the Sept7 depletion did not affect the accumulation of actomyosin at the wound edge. The cortical microtubule bundles were reduced in wound-proximal cells in Sept7 knockdown (Sept7-KD) embryos, but forced bundling of microtubules mediated by the microtubule-stabilizing protein Map7 did not rescue the Sept7-KD phenotype. Nocodazole-mediated microtubule depolymerization enhanced the Sept7-KD phenotype, suggesting that Sept7 is required for microtubule reorganization during cell elongation. Our findings indicate that septins are required for the rapid wound closure by facilitating cortical microtubule reorganization and the concentric elongation of surrounding cells.

    DOI: 10.1242/jcs.212647

    Web of Science

    PubMed

  8. 神経ネットワークの形と機能を制御する細胞骨格系分子群 Invited

    木下 専

    DOJIN BIOSCIENCE SERIES 28脳神経化学 脳はいま化学の言葉でどこまで語れるか     2018.4

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    Authorship:Lead author, Last author, Corresponding author   Language:Japanese   Publishing type:Part of collection (book)  

  9. グルタミン酸輸送体の足場となり、クリアランスを促進する シナプス周囲グリア突起内の蛋白質複合体 Invited

    木下 専

    日本生物学的精神医学会誌 (特集 精神神経疾患と興奮性アミノ酸受容体)   Vol. 28 ( 2 ) page: 84 - 86   2017.4

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    Authorship:Lead author, Last author, Corresponding author   Language:Japanese   Publishing type:Research paper (scientific journal)  

    DOI: 10.11249/jsbpjjpp.28.2_84

  10. Septin Interferes with the Temperature-Dependent Domain Formation and Disappearance of Lipid Bilayer Membranes. Reviewed International journal

    Shunsuke Yamada, Takumi Isogai, Ryugo Tero, Yohko Tanaka-Takiguchi, Toru Ujihara, Makoto Kinoshita, Kingo Takiguchi

    Langmuir : the ACS journal of surfaces and colloids   Vol. 32 ( 48 ) page: 12823 - 12832   2016.12

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    Language:English   Publishing type:Research paper (scientific journal)   Publisher:American Chemical Society ({ACS})  

    Domain formation or compartmentalization in a lipid bilayer membrane has been thought to take place dynamically in cell membranes and play important roles in the spatiotemporal regulation of their physiological functions. In addition, the membrane skeleton, which is a protein assembly beneath the cell membrane, also regulates the properties as well as the morphology of membranes because of its role as a diffusion barrier against constitutive molecules of the membrane or as a scaffold for physiological reactions. Therefore, it is important to study the relationship between lipid bilayer membranes and proteins that form the membrane skeleton. Among cytoskeletal systems, septin is unique because it forms arrays on liposomes that contain phosphoinositides, and this property is thought to contribute to the formation of the annulus in sperm flagellum. In this study, a supported lipid bilayer (SLB) was used to investigate the effect of septin on lipid bilayers because SLBs rather than liposomes are suitable for observation of the membrane domains formed. We found that SLBs containing phosphatidylinositol (PI) reversibly form domains by decreasing the temperature and that septin affects both the formation and the disappearance of the cooling-induced domain. Septin inhibits the growth of cooling-induced domains during decreases in temperature and inhibits the dispersion and the disappearance of those domains during increases in temperature. These results indicate that septin complexes, i.e., filaments or oligomers assembling on the surface of lipid bilayer membranes, can regulate the dynamics of domain formation via their behavior as an anchor for PI molecules.

    DOI: 10.1021/acs.langmuir.6b03452

    Web of Science

    PubMed

  11. Septin7 regulates inner ear formation at an early developmental stage. Reviewed International journal

    Hiroko Torii, Atsuhiro Yoshida, Tatsuya Katsuno, Takayuki Nakagawa, Juichi Ito, Koichi Omori, Makoto Kinoshita, Norio Yamamoto

    Developmental biology   Vol. 419 ( 2 ) page: 217 - 228   2016.11

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    Language:English   Publishing type:Research paper (scientific journal)  

    Septins are guanosine triphosphate-binding proteins that are evolutionally conserved in all eukaryotes other than plants. They function as multimeric complexes that interact with membrane lipids, actomyosin, and microtubules. Based on these interactions, septins play essential roles in the morphogenesis and physiological functions of many mammalian cell types including the regulation of microtubule stability, vesicle trafficking, cortical rigidity, planar cell polarity, and apoptosis. The inner ear, which perceives auditory and equilibrium sensation with highly differentiated hair cells, has a complicated gross morphology. Furthermore, its development including morphogenesis is dependent on various molecular mechanisms, such as apoptosis, convergent extension, and cell fate determination. To determine the roles of septins in the development of the inner ear, we specifically deleted Septin7 (Sept7), the non-redundant subunit in the canonical septin complex, in the inner ear at different times during development. Foxg1Cre-mediated deletion of Sept7, which achieved the complete knockout of Sept7 within the inner ear at E9.5, caused cystic malformation of inner ears and a reduced numbers of sensory epithelial cells despite the existence of mature hair cells. Excessive apoptosis was observed at E10.5,E11.5 and E12.5 in all inner ear epithelial cells and at E10.5 and E11.5 in prosensory epithelial cells of the inner ears of Foxg1Cre;Septin7floxed/floxed mice. In contrast with apoptosis, cell proliferation in the inner ear did not significantly change between control and mutant mice. Deletion of Sept7 within the cochlea at a later stage (around E15.5) with Emx2Cre did not result in any apparent morphological anomalies observed in Foxg1Cre;Septin7floxed/floxed mice. These results suggest that SEPT7 regulates gross morphogenesis of the inner ear and maintains the size of the inner ear sensory epithelial area and exerts its effects at an early developmental stage of the inner ear.

    DOI: 10.1016/j.ydbio.2016.09.012

    Web of Science

    PubMed

  12. A septin requirement differentiates autonomous and contact-facilitated T cell proliferation. Reviewed International coauthorship International journal

    Adriana M Mujal, Julia K Gilden, Audrey Gérard, Makoto Kinoshita, Matthew F Krummel

    Nature Immunology   Vol. 17 ( 3 ) page: 315 - 22   2016.3

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    Language:English   Publishing type:Research paper (scientific journal)   Publisher:Springer Nature  

    T cell proliferation is initiated by T cell antigen receptor (TCR) triggering, soluble growth factors or both. In characterizing T cells lacking the septin cytoskeleton, we found that successful cell division has discrete septin-dependent and septin-independent pathways. Septin-deficient T cells failed to complete cytokinesis when prompted by pharmacological activation or cytokines. In contrast, cell division was not dependent on septins when cell-cell contacts, such as those with antigen-presenting cells, provided a niche. This septin-independent pathway was mediated by phosphatidylinositol-3-OH kinase activation through a combination of integrins and costimulatory signals. We were able to differentiate between cytokine- and antigen-driven expansion in vivo and thus show that targeting septins has strong potential to moderate detrimental bystander or homeostatic cytokine-driven proliferation without influencing expansion driven by conventional antigen-presentation.

    DOI: 10.1038/ni.3330

    Web of Science

    PubMed

  13. Facilitation of axon outgrowth via a Wnt5a-CaMKK-CaMKIα pathway during neuronal polarization Reviewed

    MAKOTO KINOSHITA

    Molecular Brain   Vol. 9 ( 8 )   2016.1

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    Language:English   Publishing type:Research paper (scientific journal)   Publisher:BIOMED CENTRAL LTD  

    Background: Wnt5a, originally identified as a guidance cue for commissural axons, activates a non-canonical pathway critical for cortical axonal morphogenesis. The molecular signaling cascade underlying this event remains obscure.
    Results: Through Ca2+ imaging in acute embryonic cortical slices, we tested if radially migrating cortical excitatory neurons that already bore primitive axons were sensitive to Wnt5a. While Wnt5a only evoked brief Ca2+ transients in immature neurons present in the intermediate zone (IZ), Wnt5a-induced Ca2+ oscillations were sustained in neurons that migrated out to the cortical plate (CP). We wondered whether this early Wnt5a-Ca2+ signaling during neuronal polarization has a morphogenetic consequence. During transition from round to polarized shape, Wnt5a administration to immature cultured cortical neurons specifically promoted axonal, but not dendritic, outgrowth. Pharmacological and genetic inhibition of the CaMKK-CaMKI alpha pathway abolished Wnt5a-induced axonal elongation, and rescue of CaMKI alpha in CaMKI alpha-knockdown neurons restored Wnt5a-mediated axon outgrowth.
    Conclusions: This study suggests that Wnt5a activates Ca2+ signaling during a neuronal morphogenetic time window when axon outgrowth is critically facilitated. Furthermore, the CaMKK-CaMKI alpha cascade is required for the axonal growth effect of Wnt5a during neuronal polarization.

    DOI: 10.1186/s13041-016-0189-3

    Web of Science

    PubMed

  14. Methods for immunoblot detection and electron microscopic localization of septin subunits in mammalian nervous systems Invited

    L. K. Parajuli, N. Ageta-Ishihara, H. Ageta, Y. Fukazawa, M. Kinoshita

    Methods in Cell Biology   Vol. 136   page: 285 - 294   2016

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    Authorship:Last author, Corresponding author   Language:English   Publishing type:Part of collection (book)   Publisher:ELSEVIER ACADEMIC PRESS INC  

    The minimal functional units of the mammalian septin system are diverse heterooligomers of SEPT1 1-14 subunits, which are most abundantly and differentially expressed in post mitotic neurons and glia. The subunit compositions of such heterooligomers are thought to differentiate their affinity for other proteins and lipids, and subcellular localization. Thus, high-precision quantification and mapping of each subunit is necessary to understand their subcellular functions and physiological roles. However, systematic information on the localization of individual septin subunits in the mammalian nervous system is limited. Here, we present our experimental workflows for the study of septin expression and localization in the rodent brain by immunoblot and serial section immunoelectron microscopy. Our protocols, based on standard methods, have been rigorously optimized and simplified for universality and reproducibility to aid non-experts in the field.

    DOI: 10.1016/bs.mcb.2016.04.021

    Web of Science

  15. CDC42EP4/BORG4-septin complex beneath perisynaptic membrane domains of Bergmann glial processes facilitates GLAST-mediated glutamate clearance and motor learning.

    Ageta-Ishihara N., Kinoshita M.

    Molecular Biology of the Cell   Vol. 27   2016

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    Authorship:Last author, Corresponding author   Language:English   Publishing type:Research paper (international conference proceedings)  

    Web of Science

  16. A CDC42EP4/septin-based perisynaptic glial scaffold facilitates glutamate clearance. Reviewed International coauthorship International journal

    Natsumi Ageta-Ishihara, Maya Yamazaki, Kohtarou Konno, Hisako Nakayama, Manabu Abe, Kenji Hashimoto, Tomoki Nishioka, Kozo Kaibuchi, Satoko Hattori, Tsuyoshi Miyakawa, Kohichi Tanaka, Fathul Huda, Hirokazu Hirai, Kouichi Hashimoto, Masahiko Watanabe, Kenji Sakimura, Makoto Kinoshita

    Nature Communications   Vol. 6 ( 10090 ) page: 10090 - 10090   2015.12

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    Authorship:Last author, Corresponding author   Language:English   Publishing type:Research paper (scientific journal)   Publisher:Springer Nature  

    The small GTPase-effector proteins CDC42EP1-5/BORG1-5 interact reciprocally with CDC42 or the septin cytoskeleton. Here we show that, in the cerebellum, CDC42EP4 is exclusively expressed in Bergmann glia and localizes beneath specific membrane domains enwrapping dendritic spines of Purkinje cells. CDC42EP4 forms complexes with septin hetero-oligomers, which interact with a subset of glutamate transporter GLAST/EAAT1. In Cdc42ep4(-/-) mice, GLAST is dissociated from septins and is delocalized away from the parallel fibre-Purkinje cell synapses. The excitatory postsynaptic current exhibits a protracted decay time constant, reduced sensitivity to a competitive inhibitor of the AMPA-type glutamate receptors (γDGG) and excessive baseline inward current in response to a subthreshold dose of a nonselective inhibitor of the glutamate transporters/EAAT1-5 (DL-TBOA). Insufficient glutamate-buffering/clearance capacity in these mice manifests as motor coordination/learning defects, which are aggravated with subthreshold DL-TBOA. We propose that the CDC42EP4/septin-based glial scaffold facilitates perisynaptic localization of GLAST and optimizes the efficiency of glutamate-buffering and clearance.

    DOI: 10.1038/ncomms10090

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    PubMed

  17. Transgenic supplementation of SIRT1 fails to alleviate acute loss of nigrostriatal dopamine neurons and gliosis in a mouse model of MPTP-induced parkinsonism [v1; ref status: awaiting peer review, http://f1000r.es/5a9] Reviewed

    Yasuko Kitao, Natsumi Ageta-Ishihara, Ryosuke Takahashi, Makoto Kinoshita, Osamu Hori

    F1000Research   Vol. 4 ( 130 ) page: -   2015.5

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    Authorship:Last author, Corresponding author   Language:English   Publishing type:Research paper (scientific journal)   Publisher:Faculty of 1000 Ltd  

    Background Dopamine (DA) neuron-selective uptake and toxicity of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) causes parkinsonism in humans. Loss of DA neurons via mitochondrial damage and oxidative stress is reproduced by systemic injection of MPTP in animals, which serves as models of parkinsonism and Parkinson's disease (PD). This study aimed to test whether pan-neural supplementation of the longevity-related, pleiotropic deacetylase SIRT1, which confers partial tolerance to at least three models of stroke and neurodegeneration, could also alleviate MPTP-induced acute pathological changes in nigrostriatal DA neurons and neighboring glia. Results We employed a line of prion promoter-driven Sirt1-transgenic (Sirt1Tg) mice that chronically overexpress murine SIRT1 in the brain and spinal cord. Sirt1Tg and wild-type (WT) male littermates (3-4 months old) were subjected to intraperitoneal injection of MPTP. Acute histopathological changes in the midbrain and striatum (caudoputamen) were assessed with serial coronal sections triply labeled for tyrosine hydroxylase (TH), glial fibrillary acidic protein (GFAP), and nuclear DNA. In the substantia nigra pars compacta (SNpc) of the midbrain, the number of TH-positive neurons and the reactive gliosis were comparable between the Sirt1Tg and WT littermates. In the striatum, the relative fluorescence intensity of TH-positive nerve terminals and the level of gliosis did not differ by the genotypes. Conclusions Sirt1Tg and WT littermate mice exhibited comparable acute histopathological reactions to the systemic injection of MPTP, loss of TH-positive neurons and reactive gliosis. Thus, the genetic supplementation of SIRT1 does not confer histologically recognizable protection on nigrostriatal DA neurons against acute toxicity of MPTP.

    DOI: 10.12688/f1000research.6386.1

    Scopus

  18. SIRT1 attenuates severe ischemic damage by preserving cerebral blood flow Reviewed International coauthorship International journal

    Yorito Hattori, Yoko Okamoto, Kazuyuki Nagatsuka, Ryosuke Takahashi, Rajesh N. Kalaria, Makoto Kinoshita, Masafumi Ihara

    NeuroReport   Vol. 26 ( 3 ) page: 113 - 117   2015.2

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    Language:English   Publishing type:Research paper (scientific journal)   Publisher:Ovid Technologies (Wolters Kluwer Health)  

    Silent information regulator 2 homolog 1 (SIRT1) is a protein deacetylase that has been reported to suppress neurodegenerative and cardiovascular pathologies in model organisms. We have recently reported that SIRT1 overexpression preserves cerebral blood flow (CBF) after bilateral common carotid artery stenosis (∼50% stenosis) by the deacetylation of endothelial nitric oxide synthase. This study was designed to determine whether cerebral SIRT1 expression would be effective in a more severe model of cerebral ischemia caused by bilateral common carotid artery occlusion (BCAO) in vivo. Sirt1-overexpressing (Sirt1-Tg) mice (n=13) and their wild-type littermates (n=17) were subjected to BCAO for 10 min using microaneurysm clips. Temporal CBF changes were measured by laser speckle flowmetry before and 5, 10 min, and 2 h after BCAO. Histological evaluation of hippocampal changes was performed 7 days after BCAO. Histological findings were significantly less severe in Sirt1-Tg mice than in wild-type mice; wild-type mice showed strokes in the hippocampus, whereas Sirt1-Tg mice had minimal hippocampal damage 7 days after BCAO. Consistent with this observation, wild-type mice showed a severe reduction in CBF to ∼20-25% of the baseline level during BCAO, whereas Sirt1-Tg littermates showed significantly preserved CBF up to 45-50% of the baseline level. Our study provides evidence for the promising role of SIRT1 in protecting against cerebral global ischemia by preserving CBF and restoring the cerebrovascular reserve.

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  19. Silent Information Regulator 2 Homolog 1 Counters Cerebral Hypoperfusion Injury by Deacetylating Endothelial Nitric Oxide Synthase Reviewed International coauthorship

    Yorito Hattori, Yoko Okamoto, Takakuni Maki, Yumi Yamamoto, Naoya Oishi, Kenichi Yamahara, Kazuyuki Nagatsuka, Ryosuke Takahashi, Raj N. Kalaria, Hidenao Fukuyama, Makoto Kinoshita, Masafumi Ihara

    STROKE   Vol. 45 ( 11 ) page: 3403 - 3411   2014.11

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    Background and Purpose Silent information regulator 2 homolog 1 (SIRT1) is a protein deacetylase that has been reported to suppress neurodegenerative and cardiovascular diseases in model organisms. We hypothesized that neurovascular protection is one of the diverse actions of SIRT1. This study was designed to determine whether SIRT1 protects against the consequences of cerebral hypoperfusion in vivo.
    MethodsSirt1-overexpressing (Sirt1-Tg) mice driven by a prion promoter and their wild-type littermates were subjected to bilateral common carotid artery stenosis using external microcoils. Using Sirt1-Tg mice, we assessed the effect of SIRT1 on cerebral blood flow, cerebral angioarchitecture, histological and ultrastructural changes, and spatial working memory at several time points. We also evaluated the effects of preadministration of SIRT1 inhibitors or endothelial nitric oxide synthase inhibitors on cerebral blood flow after bilateral common carotid artery stenosis in Sirt1-Tg mice. Levels of acetylated and nonacetylated endothelial nitric oxide synthase were measured semiquantitatively with immunoblotting.
    Results Cerebral hypoperfusion induced by bilateral common carotid artery stenosis caused memory impairment and histological changes in wild-type littermates. However, these phenotypes were rescued in Sirt1-Tg mice, where cerebral blood flow was maintained even poststenosis. Electron microscopic analyses showed irregularities in the vascular endothelia, such as tight junction openings in wild-type mice, which were absent in Sirt1-Tg littermates. Brain endothelial nitric oxide synthase was acetylated after cerebral hypoperfusion in wild-type littermates but remained unacetylated in Sirt1-Tg mice. Moreover, treatment with SIRT1 inhibitors and endothelial nitric oxide synthase inhibitors abolished the vasculoprotective effects of SIRT1.
    Conclusions Our results indicate that neurovascular endothelial SIRT1 potentiation upregulates the nitric oxide system and counters cerebral hypoperfusion injury. This novel cerebral blood flow-preserving mechanism offers potential molecular targets for future therapeutic intervention.

    DOI: 10.1161/STROKEAHA.114.006265

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  20. Genetic deletion of SEPT7 reveals a cell type-specific role of septins in microtubule destabilization for the completion of cytokinesis. Reviewed International coauthorship International journal

    Manoj B Menon, Akihiro Sawada, Anuhar Chaturvedi, Pooja Mishra, Karin Schuster-Gossler, Melanie Galla, Axel Schambach, Achim Gossler, Reinhold Förster, Michael Heuser, Alexey Kotlyarov, Makoto Kinoshita, Matthias Gaestel

    PLoS Genetics   Vol. 10 ( 8 ) page: e1004558   2014.8

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    Cytokinesis terminates mitosis, resulting in separation of the two sister cells. Septins, a conserved family of GTP-binding cytoskeletal proteins, are an absolute requirement for cytokinesis in budding yeast. We demonstrate that septin-dependence of mammalian cytokinesis differs greatly between cell types: genetic loss of the pivotal septin subunit SEPT7 in vivo reveals that septins are indispensable for cytokinesis in fibroblasts, but expendable in cells of the hematopoietic system. SEPT7-deficient mouse embryos fail to gastrulate, and septin-deficient fibroblasts exhibit pleiotropic defects in the major cytokinetic machinery, including hyperacetylation/stabilization of microtubules and stalled midbody abscission, leading to constitutive multinucleation. We identified the microtubule depolymerizing protein stathmin as a key molecule aiding in septin-independent cytokinesis, demonstrated that stathmin supplementation is sufficient to override cytokinesis failure in SEPT7-null fibroblasts, and that knockdown of stathmin makes proliferation of a hematopoietic cell line sensitive to the septin inhibitor forchlorfenuron. Identification of septin-independent cytokinesis in the hematopoietic system could serve as a key to identify solid tumor-specific molecular targets for inhibition of cell proliferation.

    DOI: 10.1371/journal.pgen.1004558

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  21. Phosphatidic Acid ( PA)-preferring Phospholipase A1 Regulates Mitochondrial Dynamics Reviewed International coauthorship

    Takashi Baba, Yuriko Kashiwagi, Nagisa Arimitsu, Takeshi Kogure, Ayumi Edo, Tomohiro Maruyama, Kazuki Nakao, Hiroki Nakanishi, Makoto Kinoshita, Michael A. Frohman, Akitsugu Yamamoto, Katsuko Tani

    JOURNAL OF BIOLOGICAL CHEMISTRY   Vol. 289 ( 16 ) page: 11497 - 11511   2014.4

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    Background: Phosphatidic acid (PA) is involved in membrane dynamics. Results: PA-preferring phospholipase A(1) (PA-PLA(1)) affects mitochondrial morphology in an activity-dependent manner. Gene disruption of PA-PLA(1) in mice causes sperm malformation due to mitochondrial organization defects. Conclusion: PA-PLA(1) regulates mitochondrial dynamics. Significance: We demonstrate an in vivo function of PA-PLA(1) and suggest a possible mechanism of PA regulation of the mitochondrial membrane.
    Recent studies have suggested that phosphatidic acid (PA), a cone-shaped phospholipid that can generate negative curvature of lipid membranes, participates in mitochondrial fusion. However, precise mechanisms underling the production and consumption of PA on the mitochondrial surface are not fully understood. Phosphatidic acid-preferring phospholipase A(1) (PA-PLA(1))/DDHD1 is the first identified intracellular phospholipase A(1) and preferentially hydrolyzes PA in vitro. Its cellular and physiological functions have not been elucidated. In this study, we show that PA-PLA(1) regulates mitochondrial dynamics. PA-PLA(1), when ectopically expressed in HeLa cells, induced mitochondrial fragmentation, whereas its depletion caused mitochondrial elongation. The effects of PA-PLA(1) on mitochondrial morphology appear to counteract those of MitoPLD, a mitochondrion-localized phospholipase D that produces PA from cardiolipin. Consistent with high levels of expression of PA-PLA(1) in testis, PA-PLA(1) knock-out mice have a defect in sperm formation. In PA-PLA(1)-deficient sperm, the mitochondrial structure is disorganized, and an abnormal gap structure exists between the middle and principal pieces. A flagellum is bent at that position, leading to a loss of motility. Our results suggest a possible mechanism of PA regulation of the mitochondrial membrane and demonstrate an in vivo function of PA-PLA(1) in the organization of mitochondria during spermiogenesis.

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  22. SIRT1 overexpression ameliorates a mouse model of SOD1-linked amyotrophic lateral sclerosis via HSF1/HSP70i chaperone system Reviewed

    Seiji Watanabe, Natsumi Ageta-Ishihara, Shinji Nagatsu, Keizo Takao, Okiru Komine, Fumito Endo, Tsuyoshi Miyakawa, Hidemi Misawa, Ryosuke Takahashi, Makoto Kinoshita, Koji Yamanaka

    Molecular Brain   Vol. 7 ( 1 ) page: 62 - 62   2014

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    Background: Dominant mutations in superoxide dismutase 1 (SOD1) cause degeneration of motor neurons in a subset of inherited amyotrophic lateral sclerosis (ALS). The pathogenetic process mediated by misfolded and/or aggregated mutant SOD1 polypeptides is hypothesized to be suppressed by protein refolding. This genetic study is aimed to test whether mutant SOD1-mediated ALS pathology recapitulated in mice could be alleviated by overexpressing a longevity-related deacetylase SIRT1 whose substrates include a transcription factor heat shock factor 1 (HSF1), the master regulator of the chaperone system.
    Results: We established a line of transgenic mice that chronically overexpress SIRT1 in the brain and spinal cord. While inducible HSP70 (HSP70i) was upregulated in the spinal cord of SIRT1 transgenic mice (PrP-Sirt1), no neurological and behavioral alterations were detected. To test hypothetical benefits of SIRT1 overexpression, we crossbred PrP-Sirt1 mice with two lines of ALS model mice: A high expression line that exhibits a severe phenotype (SOD1(G93A)-H) or a low expression line with a milder phenotype (SOD1(G93A)-L). The Sirt1 transgene conferred longer lifespan without altering the time of symptomatic onset in SOD1(G93A)-L. Biochemical analysis of the spinal cord revealed that SIRT1 induced HSP70i expression through deacetylation of HSF1 and that SOD1(G93A)-L/PrP-Sirt1 double transgenic mice contained less insoluble SOD1 than SOD1(G93A)-L mice. Parallel experiments showed that Sirt1 transgene could not rescue a more severe phenotype of SOD1(G93A)-H transgenic mice partly because their HSP70i level had peaked out.
    Conclusions: The genetic supplementation of SIRT1 can ameliorate a mutant SOD1-linked ALS mouse model partly through the activation of the HSF1/HSP70i chaperone system. Future studies shall include testing potential benefits of pharmacological enhancement of the deacetylation activity of SIRT1 after the onset of the symptom.

    DOI: 10.1186/s13041-014-0062-1

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  23. Septins promote dendrite and axon development by negatively regulating microtubule stability via HDAC6-mediated deacetylation. Reviewed International coauthorship

    Ageta-Ishihara N, Miyata T, Ohshima C, Watanabe M, Sato Y, Hamamura Y, Higashiyama T, Mazitschek R, Bito H, Kinoshita M.

    Nature Communications   Vol. 4 ( 2532 )   2013.10

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    Neurite growth requires two guanine nucleotide-binding protein polymers of tubulins and septins. However, whether and how those cytoskeletal systems are coordinated was unknown. Here we show that the acute knockdown or knockout of the pivotal septin subunit SEPT7 from cerebrocortical neurons impairs their interhemispheric and cerebrospinal axon projections and dendritogenesis in perinatal mice, when the microtubules are severely hyperacetylated. The resulting hyperstabilization and growth retardation of microtubules are demonstrated in vitro. The phenotypic similarity between SEPT7 depletion and the pharmacological inhibition of α-tubulin deacetylase HDAC6 reveals that HDAC6 requires SEPT7 not for its enzymatic activity, but to associate with acetylated α-tubulin. These and other findings indicate that septins provide a physical scaffold for HDAC6 to achieve efficient microtubule deacetylation, thereby negatively regulating microtubule stability to an optimal level for neuritogenesis. Our findings shed light on the mechanisms underlying the HDAC6-mediated coupling of the two ubiquitous cytoskeletal systems during neural development.

    DOI: 10.1038/ncomms3532

  24. Chronic overload of SEPT4, a parkin substrate that aggregates in Parkinson's disease, causes behavioral alterations but not neurodegeneration in mice. Reviewed International journal

    Natsumi Ageta-Ishihara, Hodaka Yamakado, Takao Morita, Satoko Hattori, Keizo Takao, Tsuyoshi Miyakawa, Ryosuke Takahashi, Makoto Kinoshita

    Molecular Brain   Vol. 6 ( 1 ) page: 35 - 35   2013.8

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    BACKGROUND: In autosomal recessive early-onset Parkinsonism (PARK2), the pathogenetic process from the loss of function of a ubiquitin ligase parkin to the death of dopamine neurons remains unclear. A dominant hypothesis attributes the neurotoxicity to accumulated substrates that are exempt from parkin-mediated degradation. Parkin substrates include two septins; SEPT4/CDCrel-2 which coaggregates with α-synuclein as Lewy bodies in Parkinson's disease, and its closest homolog SEPT5/CDCrel-1/PNUTL1 whose overload with viral vector can rapidly eliminate dopamine neurons in rats. However, chronic effects of pan-neural overload of septins have never been examined in mammals. To address this, we established a line of transgenic mice that express the largest gene product SEPT4(54kDa) via the prion promoter in the entire brain. RESULTS: Histological examination and biochemical quantification of SEPT4-associated proteins including α-synuclein and the dopamine transporter in the nigrostriatal dopamine neurons found no significant difference between Sept4(Tg/+) and wild-type littermates. Thus, the hypothetical pathogenicity by the chronic overload of SEPT4 alone, if any, is insufficient to trigger neurodegenerative process in the mouse brain. Intriguingly, however, a systematic battery of behavioral tests revealed unexpected abnormalities in Sept4(Tg/+) mice that include consistent attenuation of voluntary activities in distinct behavioral paradigms and altered social behaviors. CONCLUSIONS: Together, these data indicate that septin dysregulations commonly found in postmortem human brains with Parkinson's disease, schizophrenia and bipolar disorders may be responsible for a subset of behavioral abnormalities in the patients.

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  25. セプチンによるドーパミン神経伝達機構の制御 Invited

    木下 専、田(石原)奈津実

    脳21 特集 トランスポートソーム:トランスポーター・チャネル群の相互作用に基づく神経系の機能構築   Vol. 16 ( 3 ) page: 40 - 45   2013.7

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  26. 運動による脳の活性化のしくみ Invited Reviewed

    Ageta-Ishihara N, Morita T, Yamauchi Y, Kinoshita M

    健康医科学   Vol. 28   page: 44 - 51   2013.3

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  27. Localization of septin proteins in the mouse cochlea. Reviewed International coauthorship International journal

    Atsuhiro Yoshida, Norio Yamamoto, Makoto Kinoshita, Noboru Hiroi, Takeshi Hiramoto, Gina Kang, William S Trimble, Kenji Tanigaki, Takayuki Nakagawa, Juichi Ito

    Hearing Research   Vol. 289 ( 1-2 ) page: 40 - 51   2012.7

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    Septins are a family of GTP binding proteins that are well conserved in eukaryotic species except plants. Septins contribute to the lateral compartmentalization of membranes, cortical rigidity, and the regulation of membrane trafficking by associating with membrane lipids, actin, and microtubules. The organ of Corti in the cochlea has pivotal roles in auditory perception and includes two kinds of highly polarized cells, hair and supporting cells, both of which are rich in actin and microtubules. To identify the roles of septins in the cochlea, we analyzed the localization of three septin proteins, septin 4 (SEPT4), septin 5 (SEPT5), and septin 7 (SEPT7) that are abundantly expressed in brain tissues, and also examined auditory functions of Sept4 and Sept5 null mice. SEPT4, SEPT5, and SEPT7 were expressed in inner and outer pillar cells and Deiters' cells but the distribution patterns of each protein in Deiters' cells were different. SEPT4 and SEPT7 were expressed in the phalangeal process where SEPT5 was not detected. In addition to these cells SEPT5 and SEPT7 were co-localized with presynaptic vesicles of efferent nerve terminals. Only SEPT7 was expressed in the cochlea at embryonic stages. Although expression patterns of septin proteins suggested their important roles in the function of the cochlea, both Sept4 and Sept5 null mice had similar auditory functions to their wild type littermates. Immunohistochemical analysis of Sept4 null mice showed that compensatory expression of SEPT5 in the phalangeal process of Deiters' cells may have caused functional compensation of hearing ability in Sept4 null mice.

    DOI: 10.1016/j.heares.2012.04.015

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  28. [Role of septin cytoskeleton in dopaminergic neurotransmission and neurodegeneration]. Invited

    Makoto Kinoshita

    Nihon shinkei seishin yakurigaku zasshi = Japanese journal of psychopharmacology   Vol. 32 ( 1 ) page: 25 - 9   2012.2

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    Cytoskeletal polymers play pleiotropic roles in neuroglial morphogenesis, intracellular transport, organization of pre- and post-synaptic scaffolds, etc. Thus, neuroglial dysfunction and degeneration are often accompanied by abnormalities in microtubules, actin and/or intermediate filament systems. Although our understanding of an unconventional cytoskeletal system composed of the septin family of GTP-binding proteins is far behind, recent studies have been revealing that qualitative and/or quantitative abnormalities of septins are also associated with neurodegenerative disorders including hereditary neuralgic amyotrophy, Parkinson disease, schizophrenia and bipolar disorder. A better understanding of the physiological and pathophysiological roles of the septin system should help develop useful biomarkers and therapeutic strategies for these diseases.

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  29. セプチン Invited Reviewed

    上田(石原)奈津実、木下 専

    脳科学辞典     2012

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    DOI: 10.14931/bsd.484

  30. セプチン Invited

    上谷 大介、木下 専

    日本細胞生物学会 細胞生物学用語集     2012

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  31. Downregulation of the Wnt antagonist Dkk2 links the loss of Sept4 and myofibroblastic transformation of hepatic stellate cells. Reviewed International journal

    Atsuko Yanagida, Keiko Iwaisako, Etsuro Hatano, Kojiro Taura, Fumiaki Sato, Masato Narita, Hiromitsu Nagata, Hiroyuki Asechi, Shinji Uemoto, Makoto Kinoshita

    Biochimica et Biophysica Acta - Molecular Basis of Disease   Vol. 1812 ( 11 ) page: 1403 - 1411   2011.11

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    BACKGROUND/AIMS: Sept4, a subunit of the septin cytoskeleton specifically expressed in quiescent hepatic stellate cells (HSCs), is downregulated through transdifferentiation to fibrogenic and contractile myofibroblastic cells. Since Sept4(-/-)mice are prone to liver fibrosis, we aimed to identify the unknown molecular network underlying liver fibrosis by probing the association between loss of Sept4 and accelerated transdifferentiation of HSCs. METHODS: We compared the transcriptomes of Sept4(+/+) and Sept4(-/-) HSCs undergoing transdifferentiation by DNA microarray and quantitative reverse transcription polymerase chain reaction (RT-PCR) analysis. Because Dickkopf2 (Dkk2) gene expression is reduced in Sept4(-/-) HSCs, we tested whether supplementing Dkk2 could suppress myofibroblastic transformation of Sept4(-/-) HSCs. We tested the involvement of the canonical Wnt pathway in this process by using a lymphoid enhancer-binding factor/transcription factor-luciferase reporter assay. RESULTS: We observed consistent upregulation of Dkk2 in primary cultured HSCs and in a carbon tetrachloride liver fibrosis in mice, which was decreased in the absence of Sept4. Supplementation with Dkk2 suppressed the induction of pro-fibrotic genes (α-smooth muscle actin and 2 collagen genes) and induced an anti-fibrotic gene (Smad7) in Sept4(-/-) HSCs. In human liver specimens with inflammation and fibrosis, Dkk2 immunoreactivity appeared to be positively correlated with the degree of fibrotic changes. CONCLUSIONS: Pro-fibrotic transformation of HSCs through the loss of Sept4 is, in part, due to reduced expression of Dkk2 and its homologues, and the resulting disinhibition of the canonical Wnt pathway.

    DOI: 10.1016/j.bbadis.2011.06.015

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  32. Submembranous septins as relatively stable components of actin-based membrane skeleton. Reviewed International journal

    Akari Hagiwara, Yasuhiro Tanaka, Rie Hikawa, Nobuhiro Morone, Akihiro Kusumi, Hiroshi Kimura, Makoto Kinoshita

    Cytoskeleton (Hoboken, N.J.)   Vol. 68 ( 9 ) page: 512 - 525   2011.9

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    The cell cortex is organized by the dynamic interplay between the plasma membrane, membrane proteins, and the cytoskeleton. Despite the cortical localization of septin heteropolymers in vivo and their direct interaction with phospholipid membranes in vitro, their behavior and roles remain elusive. This study characterizes the major cortical septin assembly found in mammalian tissue culture cells by fluorescence recovery after photobleaching analysis. GFP-tagged septin subunits, which colocalized with cortical actin, exhibited slower turnover than some other cortical proteins that were analyzed (e.g., actin, syntaxin-1A and a glutamate aspartate transporter [GLAST]). Perturbation of actin turnover by cytochalasin D or jasplakinolide retarded the cortical septin turnover, while septin depletion by RNAi did not recognizably affect cortical actin turnover. These phenomena are compatibly interpreted by septins' selective association with a subset of actin-based membrane skeleton, as revealed by rapid-freeze deep-etch immuno-replica electron microscopy. We applied the assay system to test septins' presumptive scaffold function on their physiological binding partners. Septin filament destabilization by RNAi-mediated subunit depletion facilitated the turnover of GLAST, depending on the carboxyl-terminal 29 residues, while a septin filament-stabilizing drug forchlorfenuron restrained more GLAST in the unexchangeable fraction. These data indicate that cortical septin heteropolymers are components of the actin-based membrane skeleton providing scaffolds for their interacting partners probably by impeding their lateral diffusion. We predict that diverse submembranous septin clusters found in vivo may serve as scaffolds or reserve pools for specific membrane-bound proteins.

    DOI: 10.1002/cm.20528

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  33. Ultrastructural localization analysis of septins in mammalian nervous system

    Hiroyuki Kurita, Yugo Fukazawa, Natsumi Ageta-Ishihara, Ryuichi Shigemoto, Makoto Kinoshita

    Neuroscience Research   Vol. 71   2011.9

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    DOI: 10.1016/j.neures.2011.07.505

  34. トランスポートソームの組織化における細胞骨格系の役割 Reviewed

    藤原敬広、木下 専

    トランスポートソームの世界-膜輸送研究の源流から未来へ     page: 380 - 386   2011.4

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  35. Septin-dependent microtubule control during mammalian neurite outgrowth Invited Reviewed

    Molecular Biology of the Cell   Vol. 22   2011

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  36. Role of the Septin Cytoskeleton for the Functional Organization of the Plasma Membrane Invited

    Makoto Kinoshita

    WATER: THE FORGOTTEN BIOLOGICAL MOLECULE     page: 259 - 267   2011

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  37. Entrapment of intracytosolic bacteria by septin cage-like structures. Reviewed International coauthorship International journal

    Serge Mostowy, Matteo Bonazzi, Mélanie Anne Hamon, To Nam Tham, Adeline Mallet, Mickaël Lelek, Edith Gouin, Caroline Demangel, Roland Brosch, Christophe Zimmer, Anna Sartori, Makoto Kinoshita, Marc Lecuit, Pascale Cossart

    Cell host & microbe   Vol. 8 ( 5 ) page: 433 - 44   2010.11

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    Actin-based motility is used by various pathogens for dissemination within and between cells. Yet host factors restricting this process have not been identified. Septins are GTP-binding proteins that assemble as filaments and are essential for cell division. However, their role during interphase has remained elusive. Here, we report that septin assemblies are recruited to different bacteria that polymerize actin. We observed that intracytosolic Shigella either become compartmentalized in septin cage-like structures or form actin tails. Inactivation of septin caging increases the number of Shigella with actin tails and enhances cell-to-cell spread. TNF-α, a host cytokine produced upon Shigella infection, stimulates septin caging and restricts actin tail formation and cell-to-cell spread. Finally, we show that septin cages entrap bacteria targeted to autophagy. Together, these results reveal an unsuspected mechanism of host defense that restricts dissemination of invasive pathogens.

    DOI: 10.1016/j.chom.2010.10.009

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  38. Rho and Anillin-dependent Control of mDia2 Localization and Function in Cytokinesis Reviewed

    Molecular Biology of the Cell   Vol. 21 ( 18 ) page: 3193 - 3204   2010.9

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    DOI: 10.1091/mbc.E10-04-0324

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  39. [Diverse physiological functions of the septin system: the protean cytoskeleton]. Invited Reviewed

    Makoto Kinoshita

    Tanpakushitsu kakusan koso. Protein, nucleic acid, enzyme   Vol. 54 ( 9 ) page: 1150 - 8   2009.7

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  40. パーキンソン病関連遺伝子Sept4 Invited Reviewed

    猪原匡史、木下 専

    「パーキンソン病―基礎・臨床研究のアップデート」日本臨牀 76巻増刊号4(通巻1142号)   Vol. 76 ( 4 ) page: 75 - 78   2009.6

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  41. [Functions of the septin cytoskeleton and its roles in dopaminergic neurotransmission]. Invited Reviewed

    Masafumi Ihara, Makoto Kinoshita

    Brain and nerve = Shinkei kenkyu no shinpo   Vol. 61 ( 4 ) page: 419 - 28   2009.4

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    Cytoskeletal polymers are component of cellular infrastructure that are required for fundamental biological processes ranging from cell division to brain functions. Unlike the knowledge available for tubulin and actin, our understanding of unconventional cytoskeletal structures composed of GTP-binding proteins belonging to the septin family is limited, despite their ubiquity and implications in human diseases. Recent studies have revealed that septin plays unique modulatory roles as an accessory component of microtubules and the actin cytoskeleton. Morphological analyses of the mammalian brain and neural cells have revealed that septins preferentially cluster beneath the extra-synaptic membrane domains in dendritic shafts and spine necks, presynaptic terminals of major neurons, and astroglial processes. Live imaging analysis revealed that septin polymers are remarkably stable in these clusters, which may serve as local cytoskeleton and/or scaffold for the organization of specialized cortical domains in neurons and glia. This hypothesis has been supported by the hypo-dopaminergic phenotype of mice that lack the Sept4 subunit and the hyper-dopaminergic phenotype of those with excess Sept4. In these cases, the septin scaffold in the dopamine neurons is considered as a determinant of the quantity of a subset of presynaptic molecules, including tSNAREs (membrane-fusion machinery) and the dopamine transporters. This finding in mouse models is in agreement with the recent findings that qualitative and/or quantitative dysregulation of septins is involved in neurodegenerative disorders such as Parkinson disease and psychological disorders such as schizophrenia and bipolar disorder. Studies on tubulin/actin indicate that a better understanding of the septin family of proteins will improve our insight into neuropathological phenomena in neurodegenerative and psychological disorders, which may help develop diagnostic markers and therapeutic strategies for such diseases.

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    Other Link: http://orcid.org/0000-0002-7102-4048

  42. Spatiotemporal association of DNAJB13 with the annulus during mouse sperm flagellum development. Reviewed International coauthorship International journal

    Jikui Guan, Makoto Kinoshita, Li Yuan

    BMC developmental biology   Vol. 9 ( 1 ) page: 23 - 23   2009.3

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    BACKGROUND: The sperm annulus is a septin-based fibrous ring structure connecting the midpiece and the principal piece of the mammalian sperm flagellum. Although ultrastructural abnormalities and functional importance of the annulus have been addressed in Sept4-null mutant mice and a subset of human patients with asthenospermia syndrome, little is known about how the structure is assembled and positioned to the midpiece-principal piece junction during mammalian sperm flagellum development. RESULTS: By performing immunofluorescence and biochemical approaches with antibodies against DNAJB13 and an annulus constituent SEPT4, we report here a spatiotemporal association of DNAJB13 with sperm annulus during mouse sperm flagellum development. DNAJB13 co-localized with SEPT4 to the annulus, and both were first able to be detected in step 9 spermatids. As spermiogenesis proceeded, the annular DNAJB13 immunosignal increased until the annulus reached the midpiece-principal piece junction, and then gradually disappeared from it in late spermiogenesis. In contrast, the SEPT4 immunosignal was relatively unaltered, and still present on annulus of mature spermatozoa. In Sept4-null mouse spermatids lacking the annulus structure, the annulus-like DNAJB13 immunosignal was still able to be detected, albeit weaker, at the neck region of the flagella. In vitro DNAJB13 was co-localized and interacted with SEPT4 directly. CONCLUSION: The direct interaction of DNAJB13 with SEPT4 in vitro and its spatiotemporal association with the annulus during sperm flagellum development, and even its annulus-like appearance in the annulus-deficient spermatids, suggest that DNAJB13 may be involved in assembling the annulus structure and positioning it towards the midpiece-principal piece junction.

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  43. Septin-Mediated Uniform Bracing of Phospholipid Membranes Reviewed

    Yohko Tanaka-Takiguchi, Makato Kinoshita, Kingo Takiguchi

    Current Biology   Vol. 19 ( 2 ) page: 140 - 145   2009.1

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    Cell shape is determined by the interplay between the lipid bilayer and the underlying network of protein polymers [1]. We explored unknown determinants involved in cell morphogenesis as factors that transform phospholipid-based liposomes (diameter 5-20 mu m). Unlabeled giant liposomes, observed through dark-field optics, were metastable in an aqueous suspension. In contrast, liposomes robustly protruded uniform tubules immediately after the addition of a brain extract to the suspension. The tubulation reaction was greatly facilitated when the liposomes contained PIP or PIP2. Biochemical analysis of the brain extract revealed that heteromeric complexes of septins, a family of polymerizing GTP/GDP-binding proteins, are responsible for the membrane transformation. Ultrastructural analysis established that each membrane tubule (diameter 0.43 +/- 0.079 mu m) is braced by a circumferential array of septin filaments. Although submembranous septin assemblies are associated with diverse cortical morphogenesis from yeast to mammals [2-5], the biophysical basis for the septin-membrane interplay remains largely unknown. Further, there is a biochemical discrepancy between the fast septin remodeling in cells and their slow self-assembly in vitro [6, 7]. This membrane-facilitated fast septin assembly demonstrated for the first time by our unique experimental system should provide important clues to characterize these processes.

    DOI: 10.1016/j.cub.2008.12.030

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  44. Amoeboid T lymphocytes require the septin cytoskeleton for cortical integrity and persistent motility. Reviewed International coauthorship International journal

    Aaron J Tooley, Julia Gilden, Jordan Jacobelli, Peter Beemiller, William S Trimble, Makoto Kinoshita, Matthew F Krummel

    Nature Cell Biology   Vol. 11 ( 1 ) page: 17 - 26   2009.1

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    The systems that refine actomyosin forces during motility remain poorly understood. Septins assemble on the T-cell cortex and are enriched at the mid-zone in filaments. Septin knockdown causes membrane blebbing, excess leading-edge protrusions and lengthening of the trailing-edge uropod. The associated loss of rigidity permits motility, but cells become uncoordinated and poorly persistent. This also relieves a previously unrecognized restriction to migration through small pores. Pharmacologically rigidifying cells counteracts this effect, and relieving cytoskeletal rigidity synergizes with septin depletion. These data suggest that septins tune actomyosin forces during motility and probably regulate lymphocyte trafficking in confined tissues.

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  45. Sept4 Invited

    祝迫惠子、木下 専

    分子細胞治療 (先端医学社)   Vol. 8 ( 2 ) page: 68 - 69   2009

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  46. 物質輸送システムの支持機構としてのセプチン系とその破綻 Invited Reviewed

    木下 専

    創薬研究者必見!最新トランスポーター研究2009 (遺伝子医学MOOK)     page: 95 - 101   2009

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  47. Septins as diagnostic markers for a subset of human asthenozoospermia. Reviewed International journal

    Yoshio Sugino, Kentaro Ichioka, Takeshi Soda, Masafumi Ihara, Makoto Kinoshita, Osamu Ogawa, Hiroyuki Nishiyama

    The Journal of Urology   Vol. 180 ( 6 ) page: 2706 - 2709   2008.12

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    PURPOSE: Septins are the major constituents of the annulus, a submembranous ring that separates the middle and principal pieces of spermatozoa. We previously reported its essential role in spermiogenesis and reproduction in mice. In the current study we investigated septin abnormality in infertile men. MATERIALS AND METHODS: Semen samples from 108 infertile patients and 21 healthy volunteers were analyzed for sperm concentration and motility. Spermatozoa were immunostained for the 2 representative septin subunits SEPT4 and SEPT7. Peripheral blood DNA from 8 patients with asthenozoospermia who had defective SEPT4 and/or SEPT7 labeling in the annuli was analyzed by direct sequencing. Clinical information and a followup review of pregnancy were obtained retrospectively from medical records. RESULTS: Specific antibodies for SEPT4 and SEPT7 consistently labeled the annuli in spermatozoa from the 21 healthy volunteers, while 14 of 108 samples (13%) from infertile patients showed defective labeling. In 33 patients with asthenozoospermia 10 samples (30%) demonstrated defective labeling for SEPT4 and/or SEPT7. We could not detect exon mutations in the SEPT4 gene by sequencing peripheral blood DNA from 8 patients with asthenozoospermia who had defective SEPT4 and/or SEPT7 labeling. During followup 8 of 14 patients (57%) with SEPT4 and/or SEPT7 labeling defects achieved successful pregnancies. CONCLUSIONS: Annulus defects were found exclusively in infertile patients. Although their prognoses do not differ from those without annulus defects, annulus labeling by septin antibodies may serve as an index for classifying a subset of spermatogenesis defects and monitoring sperm quality.

    DOI: 10.1016/j.juro.2008.08.005

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  48. Loss of Sept4 exacerbates liver fibrosis through the dysregulation of hepatic stellate cells Reviewed

    Keiko Iwaisako, Etsuro Hatano, Kojiro Taura, Akio Nakajima, Masaharu Tada, Satoru Seo, Nobuyuki Tamaki, Fumiaki Sato, Iwao Ikai, Shinji Uemoto, Makoto Kinoshita

    Journal of Hepatology   Vol. 49 ( 5 ) page: 768 - 778   2008.11

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    Background/Aims: Septins are ubiquitous and multifunctional scaffold proteins involved in cytoskeletal organization, exocytosis and other cellular processes. We disclose the quiescent hepatic stellate cells (HSCs)-specific expression of a septin subunit Sept4 in the liver, and explore the significance of the septin system in liver fibrosis.
    Methods: We analyzed the expression of alpha-smooth muscle actin (alpha-SMA), collagens and other markers in primary cultured HSCs derived from wild-type and Sept4(-/-) mice. We compared susceptibility of these mice to liver fibrosis induced by either carbon tetrachloride treatment, bile duct ligation or methionine/choline-deficient diet. Collagen deposition, the principal parameter of liver fibrosis, was quantified both histochemically (Masson's trichrome stain) and biochemically (hydroxyproline content).
    Results: In vitro, Sept4 mRNA/protein was remarkably downregulated in HSCs through myofibroblastic transformation. Sept4(-/-) HSCs showed normal morphology and proliferation, while myofibroblastic transformation as monitored by the upregulation of alpha-SMA and collagen was accelerated compared to wild-type HSCs. In vivo, liver fibrosis was consistently more severe in Sept4(-/-) mice than in wild-type littermates in all of the three paradigms of hepatitis/liver fibrosis.
    Conclusions: These data concordantly indicate that the HSC-specific septin subunit Sept4 and perhaps the septin system are involved in the suppressive modulation of myofibroblastic transformation and fibrogenesis associated with liver diseases. (C) 2008 European Association for the Study of the Liver. Published by Elsevier B.V. All rights reserved.

    DOI: 10.1016/j.jhep.2008.05.026

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  49. Structural insights shed light onto septin assemblies and function Invited Reviewed International coauthorship

    Yves Barral, Makoto Kinoshita

    Current Opinion in Cell Biology   Vol. 20 ( 1 ) page: 12 - 18   2008.2

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    While the original septin mutants were identified more than 30 years ago for their role in cytokinesis [Hartwell, LH: Genetic control of the cell division cycle in yeast. IV. Genes controlling bud emergence and cytokinesis. Exp Cell Res 1971, 69: 265-276], the architecture of septin complexes and higher order structures has remained a mystery up until very recently. Over the last few months a number of converging approaches have suddenly provided a wealth of structural information about the different levels of septin organization. Here, we review these advancements and highlight their functional consequences.

    DOI: 10.1016/j.ceb.2007.12.001

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  50. Morphogenesis of polarized epithelia requires Sept2-mediated coupling of vesicle transport to polyglutamylated microtubules. Reviewed International coauthorship

    Spiliotis ET, Hunt S, Hu Q, Kinoshita M, Nelson WJ

    Journal of Cell Biology   Vol. 180   page: 295-303   2008

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  51. Parkin deficient mice display motor disfunction and learning and memory impairment Reviewed

    Neuroscience Research   Vol. 61   page: S136 - S136   2008

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  52. Role of Septin Cytoskeleton in Spine Morphogenesis and Dendrite Development in Neurons Reviewed International coauthorship

    Tomoko Tada, Alyson Simonetta, Matthew Batterton, Makoto Kinoshita, Dieter Edbauer, Morgan Sheng

    Current Biology   Vol. 17 ( 20 ) page: 1752 - 1758   2007.10

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    Septins are GTP-binding proteins that polymerize into heteromeric filaments and form microscopic bundles or ring structures in vitro and in vivo. Because of these properties and their ability to associate with membrane, F-actin, and microtubules, septins have been generally regarded as cytoskeletal components [1, 2]. Septins are known to play roles in cytokinesis, in membrane trafficking, and as structural scaffolds; however, their function in neurons is poorly understood. Many members of the septin family, including Septin 7 (Sept7), were found by mass-spectrometry analysis of postsynaptic density (PSD) fractions of the brain [3, 4], suggesting a possible postsynaptic function of septins in neurons. We report that Sept7 is localized at the base of dendritic protrusions and at dendritic branch points in cultured hippocampal neurons-a distribution reminiscent of septin localization in the bud neck of budding yeast. Overexpression of Sept7 increased dendrite branching and the density of dendritic protrusions, whereas RNA interference (RNAi)-mediated knockdown of Sept7 led to reduced dendrite arborization and a greater proportion of immature protrusions. These data suggest that Sept7 is critical for spine morphogenesis and dendrite development during neuronal maturation.

    DOI: 10.1016/j.cub.2007.09.039

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  53. Connecting the dots between septins and the DNA damage checkpoint Invited Reviewed

    Makoto Kinoshita, Shunichi Takeda

    Cell   Vol. 130 ( 5 ) page: 777 - 779   2007.9

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    In budding yeast, septins are involved in the morphogenesis checkpoint and the DNA damage checkpoint, both of which regulate cell-cycle progression. In this issue of Cell, Kremer et al. (2007) link septins to DNA damage in mammalian cells by identifying a new signaling pathway that includes the adaptors SOCS7 and NCK. As NCK controls actin dynamics, this pathway may connect DNA damage responses and cellular morphology in metazoans.

    DOI: 10.1016/j.cell.2007.08.022

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  54. パーキンソン病の病態に迫る Invited

    猪原匡史、木下 専

    化学と生物 (日本農芸化学会誌)   Vol. 45 ( 9 ) page: 598 - 599   2007.9

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    DOI: 10.1271/kagakutoseibutsu1962.45.596

  55. Phosphorylation of adult type Sept5 (CDCrel-1) by cyclin-dependent kinase 5 inhibits interaction with syntaxin-1 Reviewed

    Makoto Taniguchi, Masato Taoka, Makoto Itakura, Akiko Asada, Taro Saito, Makoto Kinoshita, Masami Takahashi, Toshiaki Isobe, Shin-ichi Hisanaga

    Journal of Biological Chemistry   Vol. 282 ( 11 ) page: 7869 - 7876   2007.3

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    Increasing evidence implicates cyclin-dependent kinase 5 (Cdk5) in neuronal synaptic function. We searched for Cdk5 substrates in synaptosomal fractions prepared from mouse brains. Mass spectrometric analysis after two-dimensional SDS-PAGE identified several synaptic proteins phosphorylated by Cdk5-p35; one protein identified was Sept5 (CDCrel-1). Although septins were isolated originally as cell division-related proteins in yeast, Sept5 is expressed predominantly in neurons and is implicated in exocytosis. We confirmed that Sept5 is phosphorylated by Cdk5p35 in vitro and identified Ser(17) of adult type Sept5 (Sept5_v1) as a major phosphorylation site. We found that Ser17 of Sept5_v1 is phosphorylated in mouse brains. Coimmunoprecipitation from synaptosomal fractions and glutathione S-transferase-syntaxin-1A pulldown assays of Sept5_v1 expressed in COS-7 cells showed that phosphorylation of Sept5_v1 by Cdk5-p35 decreases the binding to syntaxin-1. These results indicate that the interaction of Sept5 with syntaxin-1 is regulated by the phosphorylation of Sept_v1 at Ser(17) by Cdk5-p35.

    DOI: 10.1074/jbc.M609457200

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  56. Sept4, a Component of Presynaptic Scaffold and Lewy Bodies, Is Required for the Suppression of α-Synuclein Neurotoxicity Reviewed

    Masafumi Ihara, Nobuyuki Yamasaki, Akari Hagiwara, Ai Tanigaki, Ayumi Kitano, Rie Hikawa, Hidekazu Tomimoto, Makoto Noda, Masashi Takanashi, Hideo Mori, Nobutaka Hattori, Tsuyoshi Miyakawa, Makoto Kinoshita

    Neuron   Vol. 53 ( 4 ) page: 519 - 533   2007.2

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    In Parkinson disease (PID), alpha-synuclein aggregates called Lewy bodies often involve and sequester Septin4 (Sept4), a polymerizing scaffold protein. However, the pathophysiological significance of this phenomenon is unclear. Here, we show the physiological association of Sept4 with alpha-synuclein, the dopamine transporter, and other presynaptic proteins in dopaminergic neurons; mice lacking Sept4 exhibit diminished dopaminergic neurotransmission due to scarcity of these presynaptic proteins. These data demonstrate an important role for septin scaffolds in the brain. In transgenic mice that express human alpha-synuclein(A53T) (a mutant protein responsible for familial PD), loss of Sept4 significantly enhances neuropathology and locomotor deterioration. In this PID model, insoluble deposits of Ser(129)-phosphorylated alpha-synuclein(A53T) are negatively correlated with the dosage of Sept4. In vitro, direct association with Sept4 protects alpha-synuclein against self-aggregation and Ser(129) phosphorylation. Taken together, these data show that Sept4 may be involved in PID as a dual susceptibility factor, as its insufficiency can diminish dopaminergic neurotransmission and enhance alpha-synuclein neurotoxicity.

    DOI: 10.1016/j.neuron.2007.01.019

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  57. 蛍光偏光顕微鏡が解いたセプチン線維配向の謎 Invited Reviewed

    木下 専

    蛋白質核酸酵素   Vol. 52 ( 1 )   2007.1

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  58. Small molecule inhibitor of mitotic spindle bipolarity identified in a phenotype-based screen Invited

    Kinoshita, M., Watanabe, N.

    Tanpakushitsu kakusan koso. Protein, nucleic acid, enzyme   Vol. 52 ( 13 Suppl ) page: 1796 - 1799   2007

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  59. 表現型スクリーニングで同定された紡錘体双極性の阻害剤 Invited

    木下 専、渡邊直樹

    蛋白質核酸酵素 融合発展する構造生物学とケミカルバイオロジーの最前線   Vol. 53 ( 13 ) page: 1796 - 1799   2007

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  60. 細胞分裂と細胞形態形成におけるGTP結合蛋白質セプチンの役割 Invited

    木下 専

    生化学(日本生化学会誌)   Vol. 78 ( 8 ) page: 755 - 759   2006.8

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    DOI: 10.14952/SEIKAGAKU.2021.930001

  61. Diversity of septin scaffolds Invited Reviewed

    Makoto Kinoshita

    Current Opinion in Cell Biology   Vol. 18 ( 1 ) page: 54 - 60   2006.2

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    Septins are ubiquitous GTP-binding proteins generally regarded as cytoskeletal components. Higher-order septin assemblies represented by the yeast septin collar function as cytoskeleton, providing structural support and scaffolds for many cellular factors. In metazoans, however, typical higher-order septin assemblies are often less predominant than dispersed 'low-order' septin populations. Recent studies revealed that septin populations with no obvious structure that had previously escaped our attention serve as scaffolds for kinetochore motor proteins and as sequestering depots for microtubule regulators. Unlike classic cytoskeletal polymers, which form uniform, continuous networks, septin polymers, being diverse, discontinuous and relatively static, seem suited to form discrete scaffolds. Thus, the septin system might be redefined as discrete scaffolds that are conditionally united to behave like cytoskeleton.

    DOI: 10.1016/j.ceb.2005.12.005

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  62. 再構成系と遺伝子破壊マウスによるセプチン系の解析 Invited

    木下 専

    細胞工学   Vol. 25 ( 7 ) page: 785 - 789   2006

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  63. Disruption of Sept6, a fusion partner gene of MLL, does not affect ontogeny, leukemogenesis induced by MLL-SEPT6, or phenotype induced by the loss of Sept4 Reviewed

    R Ono, M Ihara, H Nakajima, K Ozaki, Y Kataoka-Fujiwara, T Taki, K Nagata, M Inagaki, N Yoshida, T Kitamura, Y Hayashi, M Kinoshita, T Nosaka

    Molecular and Cellular Biology   Vol. 25 ( 24 ) page: 10965 - 10978   2005.12

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    Septins are evolutionarily conserved GTP-binding proteins that can heteropolymerize into filaments. Recent studies have revealed that septins are involved in not only diverse normal cellular processes but also the pathogenesis of various diseases, including cancer. SEPT6 is ubiquitously expressed in tissues and one of the fusion partner genes of MLL in the 11q23 translocations implicated in acute leukemia. However, the roles of this septin in vivo remain elusive. We have developed Sept6-deficient mice that exhibited neither gross abnormalities, changes in cytokinesis, nor spontaneous malignancy. Sept6 deficiency did not cause any quantitative changes in any of the septins evaluated in this study, nor did it cause any additional changes in the Sept4-deficient mice. Even the depletion of Sept11, a close homolog of Sept6, did not affect the Sept6-null cells in vitro, thus implying a high degree of redundancy in the septin system. Furthermore, a loss of Sept6 did not alter the phenotype of myeloproliferative disease induced by MLL-SEPT6, thus suggesting that Sept6 does not function as a tumor suppressor. To our knowledge, this is the first report demonstrating that a disruption of the translocation partner gene of MLL in 11q23 translocation does not contribute to leukemogenesis by the MLL fusion gene.

    DOI: 10.1128/MCB.25.24.10965-10978.2005

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  64. セプチン系細胞骨格と精子無力症 Invited

    西山博之、木下 専

    医学のあゆみ   Vol. 214 ( 3 ) page: 221 - 222   2005.7

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  65. A mitotic septin scaffold required for mammalian chromosome congression and segregation Reviewed International coauthorship International journal

      Vol. 307   page: 1781 - 1785   2005.3

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    DOI: 10.1126/science.1106823

  66. Cortical Organization by the Septin Cytoskeleton Is Essential for Structural and Mechanical Integrity of Mammalian Spermatozoa Reviewed

    Masafumi Ihara, Ayae Kinoshita, Shuichi Yamada, Hiromitsu Tanaka, Ai Tanigaki, Ayumi Kitano, Motohito Goto, Kazutoshi Okubo, Hiroyuki Nishiyama, Osamu Ogawa, Chiaki Takahashi, Shigeyoshi Itohara, Yoshitake Nishimune, Makoto Noda, Makoto Kinoshita

    Developmental Cell   Vol. 8 ( 3 ) page: 343 - 352   2005.3

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    Septins are polymerizing GTP binding proteins required for cortical organization during cytokinesis and other cellular processes. A mammalian septin gene Sept4 is expressed mainly in postmitotic neural cells and postmeiotic male germ cells. In mouse and human spermatozoa, SEPT4 and other septins are found in the annulus, a cortical ring which separates the middle and principal pieces. Sept4(-/-) male mice are sterile due to defective morphology and motility of the sperm flagellum. In Sept4 null spermatozoa, the annulus is replaced by a fragile segment lacking cortical material, beneath which kinesin-mediated intraflagellar transport stalls. The sterility is rescued by injection of sperm into oocytes, demonstrating that each Sept4 null spermatozoon carries an intact haploid genome. The annulus/septin ring is also disorganized in spermatozoa from a subset of human patients with asthenospermia syndrome. Thus, cortical organization based on circular assembly of the septin cytoskeleton is essential for the structural and mechanical integrity of mammalian spermatozoa.

    DOI: 10.1016/j.devcel.2004.12.005

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  67. Septins and Cytokinesis Invited Reviewed International coauthorship

    Encyclopedia of Biological Chemistry   Vol. 4   page: 22 - 26   2004.11

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    DOI: 10.1016/B0-12-443710-9/00618-9

  68. Endomucin, a sialomucin expressed in high endothelial venules, supports L-selectin-mediated rolling Reviewed

    H Kanda, T Tanaka, M Matsumoto, E Umemoto, Y Ebisuno, M Kinoshita, M Noda, R Kannagi, T Hirata, T Murai, M Fukuda, M Miyasaka

    International Immunology   Vol. 16 ( 9 ) page: 1265 - 1274   2004.9

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    Lymphocyte homing to lymph nodes is regulated by transient but specific interactions between lymphocytes and high endothelial venules (HEVs), the initial phase of which is mainly governed by the leukocyte adhesion molecule L-selectin, which recognizes sulfated and sialylated O-linked oligosaccharides displayed on sialomucin core proteins. One of the sialomucin proteins, endomucin, is predominantly expressed in vascular endothelial cells of a variety of tissues including the HEVs of lymph nodes; however, whether it functions as a ligand for L-selectin remains to be formally proven. Here we show that the endomucin splice isoform a is predominantly expressed in Ill HEVs and MAdCAM-1(+) HEVs, as seen in non-HEV-type vascular endothelial cells. Using affinity purification with soluble L-selectin, we found that HEV endomucin is specifically modified with L-selectin-reactive oligosaccharides and can bind L-selectin as well as an HEV-specific mAb, MECA-79. Our results also indicated that a 90-100 kDa endomucin species is preferentially decorated with L-selectin-reactive sugar chains, whereas an 80 kDa species represents conventional forms expressed in non-HEV-type vascular endothelial cells in lymph nodes. Furthermore, a CHO cell line expressing endomucin together with a specific combination of carbohydrate-modifying enzymes [core-2 beta-1,6-N-acetylglucosaminyltransferase (C2GnT), alpha-1,3-fucosyltransferase VII (FucTVII) and L-selectin ligand sulfotransferase (LSST)] showed L-selectin-dependent rolling under flow conditions in vitro. These results suggest that when endomucin is appropriately modified by a specific set of glycosyltransferases and a sulfotransferase, it can function as a ligand for L-selectin, and that the endomucin expressed in HEVs may represent another sialomucin ligand for L-selectin.

    DOI: 10.1093/intimm/dxh128

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  69. Chronic cerebral hypoperfusion induces white matter lesions and loss of oligodendroglia with DNA fragmentation in the rat Reviewed

    H. Tomimoto, M. Ihara, H. Wakita, R. Ohtani, J. X. Lin, I. Akiguchi, M. Kinoshita, H. Shibasaki

    Acta Neuropathologica   Vol. 106 ( 6 ) page: 527 - 534   2003.12

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    Cerebrovascular white matter lesions represent an age-related neurodegenerative condition that appears as a hyperintense signal on magnetic resonance images. These lesions are frequently observed in aging, hypertension and cerebrovascular disease, and are responsible for cognitive decline and gait disorders in the elderly population. In humans, cerebrovascular white matter lesions are accompanied by apoptosis of oligodendroglia, and have been thought to be caused by chronic cerebral ischemia. In the present study, we tested whether chronic cerebral hypoperfusion induces white matter lesions and apoptosis of oligodendroglia in the rat. Doppler flow meter analysis revealed an immediate reduction of cerebral blood flow ranging from 30% to 40% of that before operation; this remained at 52-64% between 7 and 30 days after operation. Transferrin-immunoreactive oligodendroglia decreased in number and the myelin became degenerated in the medial corpus callosum at 7 days and thereafter. Using the TUNEL method, the number of cells showing DNA fragmentation increased three- to eightfold between 3 and 30 days post-surgery compared to sham-operated animals. Double labeling with TUNEL and immunohistochemistry for markers of either astroglia or oligodendroglia showed that DNA fragmentation occurred in both of these glia. Messenger RNA for caspase-3 increased approximately twofold versus the sham-operated rats between 1 and 30 days post-surgery. Immunohistochemistry revealed up-regulation of caspase-3 in the oligodendroglia of the white matter, and also in the astroglia and neurons of the gray matter. Molecules involved in apoptotic signaling such as TNF-α and Bax were also up-regulated in glial cells. These results indicate that chronic cerebral hypoperfusion induces white matter degeneration in association with DNA fragmentation in oligodendroglia.

    DOI: 10.1007/s00401-003-0749-3

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  70. The Septins (Protein Family Review) Invited Reviewed

    Makoto Kinoshita

    Genome Biology   Vol. 4 ( 236 ) page: 1 - 9   2003.10

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    The septins make up a family of guanine-nucleotide binding proteins, most of which polymerize to form filaments. Septin genes have been found in fungi and animals but not in protozoa or plants
    yeasts have seven septin genes and humans have twelve, but Caenorhabditis elegans has only two. Some septin genes generate multiple polypeptides by alternative splicing or alternative translation start sites. Of the five conserved motifs found in other members of the GTPase superfamily, three are highly conserved in septins. Septin filaments are thought to form a cytoskeletal system that organizes higher-order structures by self-assembly and templated assembly. These multifunctional proteins are best known for their role in cytokinesis, but other functions in dividing and non-dividing cells have evolved in different lineages: budding yeast has septins specific for sporulation
    nematode septins are implicated in postembryonic morphogenesis of multiple cell lineages
    fly septins are associated with the development of germ cells, photoreceptor cells and nervous system
    and mammalian septins are implicated in exocytosis, tumorigenesis, apoptosis, synaptogenesis and neurodegeneration.

    DOI: 10.1186/gb-2003-4-11-236

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  71. Assembly of mammalian septins Invited

    Makoto Kinoshita

    Journal of Biochemistry   Vol. 134 ( 4 ) page: 491 - 496   2003.10

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    Septins are a conserved family of polymerizing guanine nucleotide binding proteins associated with diverse processes in dividing and non-dividing cells. In humans, 12 septin genes generate dozens of polypeptides, many of which comprise heterooligomeric complexes. Native and recombinant mammalian septin complexes are purified as similar to8-nm-thick filaments of variable length. Ultrastructurally, a mammalian septin filament appears an irregular array of structural segments, whose polarity is obscure. The filaments have a potential to self-assemble into higher-order structures by lateral stacking and tandem annealing, eventually forming uniformly curved bundles, i.e., rings and coils. The septin filaments also undergo templated assembly along existing actin bundles containing an adapter protein, anillin. The resultant higher-order assembly of septin filaments may provide scaffolds to recruit other molecules and/or help organize the actin-based structures. The in vitro self-assembly is an irreversible process, which is not coupled with robust nucleotide exchange or hydrolysis. In contrast, septin-based structures rearrange and disassemble in cells, which might be controlled by diverse factors (e.g., the Cdc42-borg system, anillin, syntaxin, phospholipids) and covalent modifications (e.g., phosphorylation, ubiquitination, sumoylation). An immediate goal of septin biochemistry is to define the mechanisms of assembly and disassembly of this elusive cytoskeleton.

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  72. Association of the cytoskeletal GTP-binding protein Sept4/H5 with cytoplasmic inclusions found in Parkinson's disease and other synucleinopathies Reviewed

    M Ihara, H Tomimoto, H Kitayama, Y Morioka, Akiguchi, I, H Shibasaki, M Noda, M Kinoshita

    Journal of Biological Chemistry   Vol. 278 ( 26 ) page: 24095 - 24102   2003.6

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    alpha-Synuclein-positive cytoplasmic inclusions are a pathological hallmark of several neurodegenerative disorders including Parkinson's disease, dementia with Lewy bodies, and multiple system atrophy. Here we report that Sept4, a member of the septin protein family, is consistently found in these inclusions, whereas five other septins (Sept2, Sept5, Sept6, Sept7, and Sept8) are not found in these inclusions. Sept4 and alpha-synuclein can also be co-immunoprecipitated from normal human brain lysates. When co-expressed in cultured cells, FLAG-tagged Sept4 and Myc-tagged alpha-synuclein formed detergent-insoluble complex, and upon treatment with a proteasome inhibitor, they formed Lewy body-like cytoplasmic inclusions. The tagged Sept4 and alpha-synuclein synergistically accelerated cell death induced by the proteasome inhibitor, and this effect was further enhanced by expression of another Lewy body-associated protein, synphilin-1, tagged with the V5 epitope. Moreover, co-expression of the three proteins (tagged Sept4, alpha-synuclein, and synphilin-1) was sufficient to induce cell death. These data raise the possibility that Sept4 is involved in the formation of cytoplasmic inclusions as well as induction of cell death in alpha-synuclein-associated neurodegenerative disorders.

    DOI: 10.1074/jbc.M301352200

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  73. 細胞生物学から「ケミカルジェネティクス」へ:ハーバード大学Mitchison研究室における研究の展開 Invited Reviewed

    木下 専

    現代化学 2003年2月号     page: 21 - 24   2003.1

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  74. Self- and Actin-Templated Assembly of Mammalian Septins Reviewed International coauthorship

    Makoto Kinoshita, Christine M. Field, Margaret L. Coughlin, Aaron F. Straight, Timothy J. Mitchison

    Developmental Cell   Vol. 3 ( 6 ) page: 791 - 802   2002.12

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    Septins are polymerizing GTPases required for cytokinesis and cortical organization. The principles by which they are targeted to, and assemble at, specific cell regions are unknown. We show that septins in mammalian cells switch between a linear organization along actin bundles and cytoplasmic rings, approximately 0.6 mum in diameter. A recombinant septin complex self-assembles into rings resembling those in cells. Linear organization along actin bundles was reconstituted by adding an adaptor protein, anillin. Perturbation of septin organization in cells by expression of a septin-interacting fragment of anillin or by septin depletion via siRNA causes loss of actin bundles. We conclude that septins alone self-assemble into rings, that adaptor proteins recruit septins to actin bundles, and that septins help organize these bundles.

    DOI: 10.1016/s1534-5807(02)00366-0

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  75. Mammalian septins nomenclature Reviewed International coauthorship

    IG Macara, R Baldarelli, CM Field, M Glotzer, Y Hayashi, SC Hsu, MB Kennedy, M Kinoshita, M Longtine, C Low, LJ Maltais, L McKenzie, TJ Mitchison, T Nishikawa, M Noda, EM Petty, M Peifer, Pringle, JR, PJ Robinson, D Roth, SEH Russell, H Stuhlmann, M Tanaka, T Tanaka, WS Trimble, J Ware, NJ Zeleznik-Le, B Zieger

    Molecular Biology of the Cell   Vol. 13 ( 12 ) page: 4111 - 4113   2002.12

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    There are 10 known mammalian septin genes, some of which produce multiple splice variants. The current nomenclature for the genes and gene products is very confusing, with several different names having been given to the same gene product and distinct names given to splice variants of the same gene. Moreover, some names are based on those of yeast or Drosophila septins that are not the closest homologues. Therefore, we suggest that the mammalian septin field adopt a common nomenclature system, based on that adopted by the Mouse Genomic Nomenclature Committee and accepted by the Human Genome Organization Gene Nomenclature Committee. The human and mouse septin genes will be named SEPT1-SEPT10 and Sept1-Sept10, respectively. Splice variants will be designated by an underscore followed by a lowercase "v" and a number, e.g., SEYT4_v1.

    DOI: 10.1091/mbc.E02-07-0438

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  76. Roles of septins in the mammalian cytokinesis machinery Invited Reviewed

    Kinoshita M, Noda M

    Cell Structure and Function   Vol. 26   page: 647 - 650   2002.4

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    DOI: doi.org/10.1247/csf.26.667

  77. Borg proteins control septin organization and are negatively regulated by Cdc42 Reviewed International coauthorship

    G Joberty, RR Perlungher, PJ Sheffield, M Kinoshita, M Noda, T Haystead, IG Macara

    Nature Cell Biology   Vol. 3 ( 10 ) page: 861 - 866   2001.10

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    The Cdc42 GTPase binds to numerous effector proteins that control cell polarity, cytoskeletal remodelling and vesicle transport. In many cases the signalling pathways downstream of these effectors are not known. Here we show that the Cdc42 effectors Borg1 to Borg3 bind to septin GTPases. Endogenous septin Cdc10 and Borg3 proteins can be immunoprecipitated together by an anti-Borg3 antibody. The ectopic expression of Borgs disrupts normal septin organization. Cdc42 negatively regulates this effect and inhibits the binding of Borg3 to septins. Borgs are therefore the first known regulators of mammalian septin organization and provide an unexpected link between the septin and Cdc42 GTPases.

    DOI: 10.1038/ncb1001-861

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  78. Identification of human endomucin-1 and -2 as membrane-bound O-sialoglycoproteins with anti-adhesive activity11The nucleotide sequences reported in this paper are available via GenBank accession numbers AB034694 (mouse endomucin-1), AB034695 (human endomucin-1), and AB034696 (human endomucin-2). Reviewed

    Makoto Kinoshita, Tomoyuki Nakamura, Masafumi Ihara, Tokuko Haraguchi, Yasushi Hiraoka, Kei Tashiro, Makoto Noda

    FEBS Letters   Vol. 499 ( 1-2 ) page: 121 - 126   2001.6

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    DOI: 10.1016/s0014-5793(01)02520-0

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  79. Chronic cerebral hypoperfusion induces MMP-2 but not MMP-9 expression in the microglia and vascular endothelium of white matter Reviewed

    Ihara M, Tomimoto H, Kinoshita M, Oh J, Noda M, Wakita H, Akiguchi I, Shibasaki H

    Journal of Cerebral Blood Flow and Metabolism   Vol. 21   page: 828-834   2001

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    DOI: 10.1097/00004647-200107000-00008

  80. Roles of Septins in the Mammalian Cytokinesis Machinery Reviewed

    Makoto Kinoshita, Makoto Noda

    Cell Structure and Function   Vol. 26 ( 6 ) page: 667 - 670   2001

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    DOI: 10.1247/csf.26.667

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    J-GLOBAL

  81. Phosphorylation of a New Brain-specific Septin, G-septin, by cGMP-dependent Protein Kinase Reviewed International coauthorship

    Jing Xue, Xin Wang, Chandra S. Malladi, Makoto Kinoshita, Peter J. Milburn, Imre Lengyel, John A. P. Rostas, Phillip J. Robinson

    Journal of Biological Chemistry   Vol. 275 ( 14 ) page: 10047 - 10056   2000.3

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    The septins are a family of GTPase enzymes, some of which are required for the cytokinesis stage of cell di vision and others of which are associated with exocytosis. me purified and cloned the cDNA for a 40-kDa protein from rat brain that is a substrate for type I cGMP-dependent protein kinase (PKG). The amino acid sequences of two tryptic peptides of P40 showed high homology to the septins. Molecular cloning revealed the 358-amino acid P40 to be a new member of the septin family. P40 was named G-septin, as it is phosphorylated in, vitro by PKG, but relatively poorly by the related cAMP dependent protein kinase and not by protein kinase C, Two splice variants of G-septin (alpha and beta) were found with distinct N and C termini, but a common GTPase domain. G-septin lacks the C-terminal coiled-coil domain characteristic of all other mammalian septins and uniquely has two predicted phosphorylation site motifs for type I PKG. Photoaffinity labeling with [alpha-P-32]GTP confirmed that G-septin is a GTP-binding protein. Northern blotting showed that G-septin mRNA (5.0 kilobases) is highly expressed in brain and undetectable in 12 other tissues, indicating that the G-septins are primarily neuronal proteins. Very low levels of 6.0-, 3.4-, and 2.6-kilobase transcripts were found in testis. Our results reveal a new class of brain-specific septins that may be regulated by PKG in neurons.

    DOI: 10.1074/jbc.275.14.10047

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  82. Differential localization of septins in the mouse brain Reviewed

    Ayae Kinoshita, Makoto Noda, Makoto Kinoshita

    Journal of Comparative Neurology   Vol. 428 ( 2 ) page: 223 - 239   2000

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    We have carried out a comparative immunohistochemical study on four members of the septin family, CDCrel-1, Septin6, CDC10, and H5, which are abundantly expressed in the adult mouse brain. We found that each septin showed overlapping but distinct distribution at the levels of light and electron microscopy. CDCrel-1 was abundant in inhibitory presynaptic terminals and associated with GABAergic vesicles in the thalamus, globus pallidus, and cerebellar nuclei. Septin6 was associated with synaptic vesicles in various brain regions, including glomeruli of the olfactory bulb. CDC10 was diffusely expressed in the brain and was localized beneath presynaptic membrane and astroglial processes. H5 was localized in the astroglial processes in some specific brain regions. The differential expression and subcellular localization of these septins indicates that a given neuron or glial cell expresses a specific set of septin monomers and that the resulting septin complexes with distinct compositions may play distinct roles in the brain. (C) 2000 Wiley-Liss, Inc.

    DOI: 10.1002/1096-9861(20001211)428:2<223::aid-cne3>3.0.co;2-m

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  83. Identification of Septins in Neurofibrillary Tangles in Alzheimer's Disease Reviewed

    Ayae Kinoshita, Makoto Kinoshita, Haruhiko Akiyama, Hidekazu Tomimoto, Ichiro Akiguchi, Sharad Kumar, Makoto Noda, Jun Kimura

    The American Journal of Pathology   Vol. 153 ( 5 ) page: 1551 - 1560   1998.11

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    Septins are evolutionarily conserved cytoskeletal GTPases that can form heteropolymer complexes involved hi cytokinesis and other cellular processes. We detected expression of the human septin genes Nedd5, H5, Diff6, and hCDC10 in postmortem brain tissues using the reverse transcription-coupled polymerase chain reaction and their products by immunoblot analysis, Four antibodies directed against three septins, Nedd5, H5, and Diff6, consistently labeled neurofibrillary tangles, neuropil threads, and dystrophic neurites in the senile plaques in brains affected by Alzheimer's disease but did not label obvious structures in young control brains. Immunoelectron microscopy revealed that Nedd5 localized to the paired helical filaments, Pre-tangles, the precursory granular deposits that accumulate in the neuronal cytoplasm, also were labeled with the antibodies. These findings suggest that at least the three septins are associated with tau-based paired helical filament core, and may contribute to the formation of neurofibrillary tangle as integral constitutents of paired helical filaments.

    DOI: 10.1016/s0002-9440(10)65743-4

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  84. Caspases in Cell Death Reviewed International coauthorship

    Loretta Dorstyn, Makoto Kinoshita, Sharad Kumar

    Results and Problems in Cell Differentiation   Vol. 24   page: 1 - 24   1998

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    DOI: 10.1007/978-3-540-69185-3_1

  85. cDNA Cloning, Expression Analysis, and Mapping of the MouseNedd4Gene Reviewed International coauthorship

    Sharad Kumar, Kieran F. Harvey, Makoto Kinoshita, Neal G. Copeland, Makoto Noda, Nancy A. Jenkins

    Genomics   Vol. 44 ( 1 ) page: 156 - 156   1997.8

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    DOI: 10.1006/geno.1997.4882

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  86. A novel gene encoding a ferredoxin reductase-like protein expressed in the neuroectoderm in Xenopus neurula Reviewed

    Seigo Hatada, Makoto Kinoshita, Hirofumi Sakumoto, Reina Nishihara, Makoto Noda, Makoto Asashima

    Gene   Vol. 194 ( 2 ) page: 297 - 299   1997.7

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    In an attempt to elucidate the molecular mechanisms of early neural development in Xenopus laevis, we identified, using a differential display method, several genes that are induced after Concanavalin A treatment in the animal caps prepared from stage 9 blastula. One such gene was found to encode a possible type IIIa membrane protein of 66.2 kDa sharing similarities with several prokaryotic and eukaryotic redox enzymes, hence the putative product was named Nfrl, neurula-specific ferredoxin reductase-like protein. Northern blot analysis confirmed that the expression of the Nfrl gene is up-regulated around the neurula stage, and is much lower in embryos of earlier stages and in adult tissues. The temporally limited expression of this gene implies neurula- and early larva-specific redox reactions of certain substrates, the nature of which remains to be elucidated. (C) 1997 Elsevier Science B.V.

    DOI: 10.1016/s0378-1119(97)00208-4

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  87. Nedd5, a mammalian septin, is a novel cytoskeletal component interacting with actin-based structures Reviewed International coauthorship

    M Kinoshita, S Kumar, A Mizoguchi, C Ide, A Kinoshita, T Haraguchi, Y Hiraoka, M Noda

    Genes & Development   Vol. 11 ( 12 ) page: 1535 - 1547   1997.6

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    The mouse Nedd5 gene encodes a 41.5-kD GTPase similar to the Saccharomyces and Drosophila septins essential for cytokinesis. Nedd5 accumulates near the contractile ring from anaphase through telophase, and finally condenses into the midbody. Microinjection of anti-Nedd5 antibody interferes with cytokinesis, giving rise to binucleated cells. In interphase and postmitotic cells, Nedd5 localizes to fibrous or granular structures depending on the growth state of the cell. The Nedd5-containing fibers are disrupted by microinjection of GTP gamma S and by Nedd5 mutants lacking GTP-binding activity, implying that GTP hydrolysis is required for its assembly. The Nedd5-containing fibers also appear to physically contact actin bundles and focal adhesion complexes and are disrupted by cytochalasin D, C3 exoenzyme, and serum starvation, suggesting a functional interaction with the actin-based cytoskeletal systems in interphase cells.

    DOI: 10.1101/gad.11.12.1535

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  88. Up-Regulation of the Nedd2 Gene Encoding an ICE/Ced-3-Like Cysteine Protease in the Gerbil Brain After Transient Global Ischemia Reviewed International coauthorship

    Makoto Kinoshita, Hidekazu Tomimoto, Ayae Kinoshita, Sharad Kumar, Makoto Noda

    Journal of Cerebral Blood Flow & Metabolism   Vol. 17 ( 5 ) page: 507 - 514   1997.5

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    We assessed the expression of several genes encoding pro-apoptotic cysteine proteases similar to interleukin-1 beta converting enzyme (ICE) and nematode Ced-3 in association with delayed neuronal death (DND) after transient forebrain ischemia in Mongolian gerbil. The levels of the two species of Nedd2 mRNA concomitantly increased about twofold in the whole forebrain at 3-6 h after 10-min ischemia and declined to the basal level by 24 h. In situ hybridization revealed that the Nedd2 gene was up-regulated in some neuronal populations in CA(1) and CA(3) regions of the hippocampus. In contrast, expression of ICE, CPP32/Yama/Apopain, and TX/ICErelII did not change within 48 h. These observations raise the possibility that up-regulation of Nedd2 in the vulnerable neurons may contribute to the proteolytic processes preceding the manifestation of apoptosis and/or necrosis after ischemic insult.

    DOI: 10.1097/00004647-199705000-00004

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  89. Characterization of a mammalian cell death gene Nedd2 Reviewed International coauthorship

    Kumar S, Kinoshita M, Noda M

    Leukemia   Vol. 11 Suppl 3   page: 385 - 386   1997.4

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  90. cDNA Cloning, Expression Analysis, and Mapping of the Mouse Nedd4 Gene Reviewed International coauthorship

    Sharad Kumar, Kieran F. Harvey, Makoto Kinoshita, Neal G. Copeland, Makoto Noda, Nancy A. Jenkins

    Genomics   Vol. 40 ( 3 ) page: 435 - 443   1997.3

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    The Nedd4 gene was initially identified by a subtraction cloning approach as a highly expressed transcript in the mouse embryonic brain. Cloning of the Nedd4 cDNA indicated that it can encode a protein of approximately 103 kDa, consisting of a Ca2+ and phospholipid binding domain, three putative protein-protein interaction domains (the WW domains), and a carboxyl-terminus region similar to the ubiquitin-protein ligase domain (hect domain). In mouse embryos, the expression of Nedd4 in the central nervous system is highest during neurogenesis and decreases as development progresses. In addition to the central nervous system, the expression of Nedd4 is detected in various embryonic tissues and persists in most adult tissues. Using an antibody raised against a fusion protein, we show that Nedd4 protein is localized to the cellular cytoplasm. We have mapped the mouse Nedd4 gene to chromosome 9 using an interspecific backcross panel. Nedd4 maps to a previously defined homologous region between human and mouse chromosomes and thus provides additional information regarding interspecies comparative mapping. (C) 1997 Academic Press.

    DOI: 10.1006/geno.1996.4582

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  91. Origin, expression and possible functions of the two alternatively spliced forms of the mouse Nedd2 mRNA. Reviewed International coauthorship

    Kumar S, Kinoshita M, Dorstyn L, Noda M

    Cell Death and Differentiation   Vol. 4   page: 378-387   1997

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  92. An interferon regulatory factor-related gene (xIRF-6) is expressed in the posterior mesoderm during the early development of Xenopus laevis Reviewed

    Seigo Hatada, Makoto Kinoshita, Shuji Takahashi, Reina Nishihara, Hirofumi Sakumoto, Akimasa Fukui, Makoto Noda, Makoto Asashima

    Gene   Vol. 203 ( 2 ) page: 183 - 188   1997

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    Out of a Xenopus neurula cDNA library, we isolated a clone which encodes a 52.4-kDa protein highly similar to the mouse interferon regulatory factor, IRF-6, whose function is unknown. The mRNA of this gene, named xIRF-6, seems to be maternally transmitted, but its amount rapidly decreases after the tailbud stage. Whole-mount in situ hybridization showed that xIRF-6 mRNA is expressed in the presumptive semitic mesoderm in the late gastrula, and then confined to a segment of posterior somite during the neurula through the tailbud stage. The temporally and spatially limited expression of the xIRF-6 gene product may contribute to the transcriptional regulation of specific genes which are necessary for the development of the posterior somites. (C) 1997 Elsevier Science B.V.

    DOI: 10.1016/s0378-1119(97)00512-x

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  93. Origin, expression and possible functions of the two alternatively spliced forms of the mouse Nedd2 mRNA Reviewed

    Sharad Kumar, Makoto Kinoshita, Loretta Dorstyn, Makoto Noda

    Cell Death and Differentiation   Vol. 4 ( 5 ) page: 378 - 387   1997

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    The Nedd2 gene, a member of the ICE/ced-3 family, transcribes two mRNA species by differential splicing. One of these encodes a shorter truncated version of the protein termed Nedd2s/ICH-1s. While Nedd2/ICH-1 induces apoptosis, ICH-1s was proposed to be a negative regulator of cell death. In this communication we have analyzed various aspects of the Nedd2s/ICH-1s. The coding region of mouse Nedd2 consists of 10 exons and alternative splicing of the 7th exon results in the generation of two transcripts. Although Nedd2s expression conferred partial resistance to serum withdrawal-induced apoptosis in NIH-3T3 cells, it failed to prevent cell death induced by the overexpression of Nedd2 and was unable to suppress apoptosis in two other cell lines (N18 and FDC-P1) under factor deprived conditions. Our results indicate that Nedd2s/ICH-1s is not a general modulator of cell death.

    DOI: 10.1038/sj.cdd.4400251

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  94. Caspase-2/Nedd2/Ich-1 Invited Reviewed

    木下 専、野田 亮

    アポトーシス (Bio Science用語ライブラリー)     page: 105 - 106   1996.12

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  95. Identification of a Xenopus glutamine synthetase gene abundantly expressed in the embryonic nervous system but not in adult brain Reviewed

    Seigo Hatada, Makoto Kinoshita, Makoto Noda, Makoto Asashima

    FEBS Letters   Vol. 371 ( 3 ) page: 287 - 292   1995.9

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    We used a PCR-based subtraction cloning procedure with concanavalin A-treated and -untreated animal caps from stage 9 Xenopus embryos to search for genes up-regulated during early neural development. One such gene was found to encode a protein homologous to several known glutamine synthetases, and we named it xGS, Molecular hybridization studies revealed that xGS mRNA is maternally transmitted and abundantly expressed in neuroectoderm-derived tissues during the gastrula and neurula stages, The expression of xGS mRNA in the nervous system continues until the larval stages, but declines thereafter and becomes undetectable in adult brain, Considering its metabolic activity and potential neuroprotective effect against the neurotoxic substances such as glutamate and ammonia, the glutamine synthetase may play an important role in the early stages of vertebrate neural development.

    DOI: 10.1016/0014-5793(95)00913-t

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  96. HMG-X, a Xenopus gene encoding an HMG1 homolog, is abundantly expressed in the developing nervous system Reviewed

    FEBS Letters   Vol. 352 ( 2 ) page: 191 - 196   1994.9

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    DOI: 10.1016/0014-5793(94)00909-0

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  97. Induction of apoptosis by the mouse Nedd2 gene, which encodes a protein similar to the product of the Caenorhabditis elegans cell death gene ced-3 and the mammalian IL-1 beta-converting enzyme Reviewed International coauthorship

    S Kumar, M Kinoshita, M Noda, N G Copeland, N A Jenkins

    Genes & Development   Vol. 8 ( 14 ) page: 1613 - 1626   1994.7

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    Language:English   Publishing type:Research paper (scientific journal)   Publisher:COLD SPRING HARBOR LAB PRESS  

    By subtraction cloning we previously identified a set of mouse genes (named Nedd1 through Nedd10) with developmentally down-regulated expression in brain. We now show that one such gene, Nedd2, encodes a protein similar to the mammalian interleukin-1 beta-converting enzyme (ICE) and the product of the Caenorhabditis elegans cell death gene ced-3 (CED-3). Both ICE and CED-3 are known to encode putative cysteine proteases and induce apoptosis when overexpressed in cultured cells. Overexpression of Nedd2 in cultured fibroblast and neuroblastoma cells also resulted in cell death by apoptosis, which was suppressed by the expression of the human bcl-2 gene, indicating that Nedd2 is functionally similar to the ced-3 gene in C. elegans. We also show that during embryonic development, Nedd2 is highly expressed in several types of mouse tissue undergoing high rates of programmed cell death such as central nervous system and kidney. Our data suggest that Nedd2 is an important component of the mammalian programmed cell death machinery.

    DOI: 10.1101/gad.8.14.1613

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  98. Vector Analysis of Corneal Astigmatism after Scleral Buckling Surgery Reviewed

    Makoto Kinoshita, Hidenobu Tanihara, Akira Negi, Shin-Ichiro Kawano, Hitoshi Ishigouoka, Yoshiki Veda, Satomi Suzuki-Yoshida, Yoshihito Honda

    Ophthalmologica   Vol. 208 ( 5 ) page: 250 - 253   1994

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    We conducted a short-term prospective study of 125 eyes with retinal detachment to examine changes of corneal astigmatism about 2 weeks after scleral buckling surgery, using a vector method on 2-fold-angle rectangular coordinates. Segmental buckles of one to less than two quadrants produced significantly greater changes in astigmatism (1.65+/-0.97 dptr) than those of less than one quadrant (0.88+/-0.75 dptr) and those spanning two quadrants or more (1.09+/-0.38 dptr) (p=0.0005, Kruskal-wallis test; n=73). The amplitude of differential vectors after explant buckling surgery (1.33+/-0.89 dptr, n=24) was significantly greater than after implant buckling surgery (0.65+/-0.45 dptr, n=15) (p=0.009, Mann-Whitney U-test) with buckles of one quadrant or less. Differential vectors tended to direct toward the buckles. There was no obvious directional tendency in the case of encircling procedures (n=52).

    DOI: 10.1159/000310501

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  99. 癌遺伝子と神経系の発生・分化・機能 Invited Reviewed

    木下 専、野田 亮

    実験医学 増刊「脳神経系の発生・分化と可塑性」   Vol. 11 ( 10 ) page: 104 - 112   1993.6

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  100. Expression of DRG during murine embryonic development Reviewed International coauthorship

    Takashi Sazuka, Makoto Kinoshita, Yasuhiro Tomooka, Yoji Ikawa, Makoto Noda, Sharad Kumar

    Biochemical and Biophysical Research Communications   Vol. 189 ( 1 ) page: 371 - 377   1992.11

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    DOI: 10.1016/0006-291x(92)91568-b

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  101. Central serous chorioretinopathy

        1991

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Books 8

  1. セプチンによるドーパミン神経伝達機構の制御「脳21 特集 トランスポートソーム:トランスポーター・チャネル群の相互作用に基づく神経系の機能構築」

    木下 専、上田(石原)奈津実( Role: Joint author)

    京都廣川書店  2013.7 

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    Responsible for pages:40-45   Language:Japanese Book type:Scholarly book

  2. ドーパミン神経伝達と変性におけるセプチン細胞骨格系の役割

    木下 専( Role: Sole author)

    日本神経精神薬理学会誌  2012 

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    Responsible for pages:25-29   Language:Japanese Book type:Scholarly book

  3. トランスポートソームの組織化における細胞骨格系の役割

    藤原敬広、木下 専( Role: Joint author)

    トランスポートソームの世界-膜輸送研究の源流から未来へ  2011 

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    Responsible for pages:380-386   Language:Japanese Book type:Scholarly book

  4. Role of the Septin Cytoskeleton for the Functional Organization of the Plasma Membrane (In Water: The Forgotten Biological Molecule)

    Makoto Kinoshita( Role: Sole author ,  Chapter 14)

    Pan Stanford Publishing  2010 

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    Language:English Book type:Scholarly book

  5. パーキンソン病関連遺伝子Sept4

    猪原匡史、木下 専( Role: Joint author)

    日本臨床 増刊号「パーキンソン病―基礎・臨床研究のアップデート」  2009 

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    Responsible for pages:75-78   Language:Japanese Book type:Scholarly book

  6. Insight into septin functions from mouse models. (In The Septins (eds. S.E.H. Russell, J.Pringle, and P. Hall)) Reviewed

    Kinoshita M( Role: Sole author ,  Chapter 15)

    John Wiley & Sons  2008.11  ( ISBN:978-0-470-77969-9

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    Total pages:380   Responsible for pages:319-336   Language:English Book type:Scholarly book

  7. Septins and Cytokinesis (In Encyclopedia of Biological Chemistry (eds.W.J. Lennarz, M.D. Lane, D.W. Cleveland, et al.)) Reviewed International journal

    Kinoshita M, Field CM( Role: Joint author)

    Academic Press  2004 

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    Responsible for pages:22-26   Language:English Book type:Scholarly book

    DOI: 10.1016/B0-12-443710-9/00618-9

  8. Caspases in cell death. In Results and Problems in Cell Differentiation, vol. 24 Reviewed International journal

    Dorstyn L, Kinoshita M, Kumar S

    Springer-Verlag  1998 

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    Language:English Book type:Scholarly book

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MISC 29

  1. CDC42EP4/BORG4-septin complex beneath perisynaptic membrane domains of Bergmann glial processes facilitates GLAST-mediated glutamate clearance and motor learning.

    N. Ageta-Ishihara, M. Kinoshita

    MOLECULAR BIOLOGY OF THE CELL   Vol. 27   2016

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    Language:English   Publishing type:Research paper, summary (international conference)   Publisher:AMER SOC CELL BIOLOGY  

    Web of Science

  2. 「長寿遺伝子」SIRT1 はeNOS を脱アセチル化することで脳低灌流侵襲に対する抵抗性を賦与する Reviewed

    猪原匡史, 服部頼都, 山本由美, 大石直也, 長束一行, 高橋良輔, 福山秀直, 木下専

    第40回日本脳卒中学会総会     2015.3

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    Language:Japanese   Publishing type:Research paper, summary (national, other academic conference)  

  3. A POTENT PROTECTIVE ROLE OF SIRT1 AGAINST CHRONIC CEREBRAL HYPOPERFUSION Reviewed

    服部頼都, 岡本洋子, 山本由美, 大石直也, 長束一行, 高橋良輔, 福山秀直, 木下専, 猪原匡史

    第55回日本神経学会総会     2014.5

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    Language:Japanese   Publishing type:Research paper, summary (national, other academic conference)  

  4. セプチン変異マウスを用いた未知の空間学習・記憶メカニズムの探索

    上田(石原, 奈津実, 澤田明宏, 真野善有, 増田博紀, 西岡朋生, 貝淵弘三, 高雄啓三, 宮川剛, 重本隆一, 深澤有吾, 木下専

    日本分子生物学会年会プログラム・要旨集(Web)   Vol. 36th   page: WEB ONLY 2AW13-4   2013

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    Language:Japanese  

    J-GLOBAL

  5. Septins promote dendrite and axon development by negatively regulating microtubule stability via HDAC6-mediated deacetylation

    N. Ageta-Ishihara, T. Miyata, H. Bito, M. Kinoshita

    MOLECULAR BIOLOGY OF THE CELL   Vol. 24   2013

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    Language:English   Publishing type:Research paper, summary (international conference)   Publisher:AMER SOC CELL BIOLOGY  

    Web of Science

  6. SIRT1による脳虚血耐性機構に関する検討

    HATTORI YORITO, IHARA TADAFUMI, OKAMOTO YOKO, KITAMURA AKIHIRO, HASE YOSHIKI, AYAKI TAKASHI, KINOSHITA MAKOTO, OISHI NAOYA, FUKUYAMA HIDENAO, ITO HIDEFUMI, TAKAHASHI RYOSUKE

    日本神経学会学術大会プログラム・抄録集   Vol. 53rd   page: 414   2012

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    Language:Japanese  

    J-GLOBAL

  7. ドーパミン神経伝達と変性におけるセプチン細胞骨格系の役割 Invited

    木下 専

    日本神経精神薬理学会誌   Vol. 32   page: 25 - 29   2012

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    Language:Japanese   Publishing type:Article, review, commentary, editorial, etc. (scientific journal)  

  8. 慢性脳低灌流はタウオパチーモデルマウスのタウ毒性を加速する

    山田 真人, 猪原 匡史, 岡本 洋子, 伊東 秀文, 木下 専, 冨本 秀和, 高橋 良輔

    臨床神経学   Vol. 51 ( 12 ) page: 1333   2011.12

  9. Septin-dependent microtubule control during mammalian neurite outgrowth.

    N. Ageta-Ishihara, M. Kinoshita

    MOLECULAR BIOLOGY OF THE CELL   Vol. 22   2011

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    Language:English   Publishing type:Research paper, summary (international conference)   Publisher:AMER SOC CELL BIOLOGY  

    Web of Science

  10. Exploring functions of the septin cytoskeleton in the formation and remodeling of neuroglial network

    Natsumi Ageta-Ishihara, Akari Hagiwara, Hiroyuki Kurita, Takao Morita, Takashi Tomonaga, Yugo Fukazawa, Ryuichi Shigemoto, Makoto Kinoshita

    Neuroscience Research   Vol. 68   page: E124 - E124   2010.1

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    Language:English   Publisher:Elsevier {BV}  

    DOI: 10.1016/j.neures.2010.07.2121

    Web of Science

  11. セプチン細胞骨格の変幻自在な高次集合性と多彩な生理機能 Invited

    木下 専

    蛋白質核酸酵素   Vol. 54 ( 9 ) page: 1150 - 1158   2009.7

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    Publisher:共立出版  

  12. 関連遺伝子 Sept4 (パーキンソン病--基礎・臨床研究のアップデート) -- (病因) Invited

    猪原匡史, 木下 専

    日本臨床   Vol. 67   page: 75 - 78   2009.6

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    Publisher:日本臨床社  

  13. セプチン細胞骨格系の機能とドーパミン神経伝達における役割 (特集 大脳基底核--分子基盤から臨床まで) Invited

    猪原匡史, 木下 専

    Brain and nerve   Vol. 61 ( 4 ) page: 419 - 428   2009.4

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    Language:Japanese   Publisher:医学書院  

  14. 物質輸送システムの支持機構としてのセプチン系とその破綻 Invited

    木下 専

    遺伝子医学MOOK「創薬医学者必見!最新トランスポーター研究2009」 (杉山雄一、金井好克 編集)   ( 12 ) page: 95 - 101   2009

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    Language:Japanese   Publishing type:Book review, literature introduction, etc.  

  15. Parkin deficient mice display motor disfunction and learning and memory impairment

    Keiko Nishio, Nobuyuki Yamasaki, Masafumi Ihara, Makoto Kinoshita, Tsuyoshi Miyakawa, Hidekazu Tomimoto, Ryosuke Takahashi

    NEUROSCIENCE RESEARCH   Vol. 61   page: S136 - S136   2008

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    Language:English   Publishing type:Research paper, summary (international conference)   Publisher:ELSEVIER IRELAND LTD  

    Web of Science

  16. パーキンソン病の病態に迫る : とらわれていた善玉タンパク質

    猪原匡史, 木下 専

    化学と生物   Vol. 45 ( 9 ) page: 598 - 599   2007.9

  17. Sept4, a component of presynaptic scaffold and Lewy bodies, is required for the suppression of α-synuclein neurotoxicity

    Makoto Kinoshita, Masafumi Ihara, Akari Hagiwara, Nobutaka Hattori, Tsuyoshi Miyakawa

    Neuroscience Research   Vol. 58   page: S58 - S58   2007.1

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    Language:English   Publisher:Elsevier {BV}  

    DOI: 10.1016/j.neures.2007.06.341

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  18. Small molecule inhibitor of mitotic spindle bipolarity identified in a phenotype-based screen (ケミカルバイオロジー) -- (代表的論文からケミカルバイオロジーの歴史をさかのぼる 生理活性物質のケミカルバイオロジーを開拓した論文) Invited

    木下 専, 渡邊直樹

    蛋白質核酸酵素   Vol. 52 ( 13 ) page: 1796 - 1799,1509〜1510   2007

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  19. 細胞分裂と細胞形態形成におけるGTP結合タンパク質セプチンの役割

    木下専

    生化学   Vol. 78 ( 8 ) page: 755 - 759   2006.8

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    Publisher:日本生化学会  

  20. Special Review 再構成系と遺伝子破壊マウスによるセプチン系の解析:セプチン欠損マウスはなぜ潰れないのか?

    木下専

    細胞工学   Vol. 25 ( 7 ) page: 785 - 789   2006.7

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    Publisher:秀潤社  

  21. ポスドク留学記 見たり,聞いたり,感じたり(11)細胞生物学から「ケミカルジェネティクス」へ--Mitchison研究室における研究の展開

    木下専

    現代化学   ( 383 ) page: 21 - 24   2003.2

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    Publisher:東京化学同人  

  22. Self- and actin-templated assembly of mammalian septins

    M Kinoshita, CM Field, ML Coughlin, AF Straight, TJ Mitchison

    MOLECULAR BIOLOGY OF THE CELL   Vol. 13   page: 202A - 202A   2002.11

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    Language:English   Publishing type:Research paper, summary (international conference)   Publisher:AMER SOC CELL BIOLOGY  

    Web of Science

  23. Biochemical and ultrastructural analysis of the purified and reconstituted mammalian septin complexes

    M Kinoshita, CM Field, ML Coughlin, TJ Mitchison

    MOLECULAR BIOLOGY OF THE CELL   Vol. 12   page: 296A - 297A   2001.11

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    Language:English   Publishing type:Research paper, summary (international conference)   Publisher:AMER SOC CELL BIOLOGY  

    Web of Science

  24. Chronic Cerebral Hypoperfusion Induces MMP-2 but not MMP-9 Expression in the Microglia and Vascular Endothelium of White Matter

    Masafumi Ihara, Hidekazu Tomimoto, Makoto Kinoshita, Junseo Oh, Makoto Noda, Hideaki Wakita, Ichiro Akiguchi, Hiroshi Shibasaki

    Journal of Cerebral Blood Flow & Metabolism   Vol. 21   page: 828 - 834   2001.7

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    Publisher:Nature Publishing Group  

    White matter lesions are closely associated with cognitive impairment and motor dysfunction in the aged. To explore the pathophysiology of these lesions, the authors examined the expression of matrix metalloproteinase-2 (MMP-2) and MMP-9 in the white matter in a rat model of chronic cerebral hypoperfusion. After bilateral clipping of the common carotid arteries, myelin staining revealed demyelinating changes in the optic tract and the corpus callosum on day 7. Zymographic analyses indicated an increase in the level of MMP-2, but not MMP-9, after the hypoperfusion. Immunohistochemical analyses revealed the presence (most abundantly on day 3) of MMP-2-expressing activated microglia in the optic tract and corpus callosum. In contrast, the capillary endothelial cells expressed MMP-2 later. IgM-immunoreactive glial cells were absent in the sham-operated animals, but were present in the hypoperfused animals by day 3, reflecting the disrupted blood-brain barrier. These findings suggest that the main sources of the elevated MMP-2 were the microglia and the endothelium, and that these cells may contribute to the remodeling of the white matter myelin and microvascular beds in chronic cerebral hypoperfusion.

    DOI: 10.1097/00004647-200107000-00008

    Scopus

    PubMed

  25. The mammalian septin complexes: composition, ultrastructure and interaction

    M Kinoshita, CM Field, ML Coughlin, TJ Mitchison

    MOLECULAR BIOLOGY OF THE CELL   Vol. 11   page: 102A - 102A   2000.12

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    Web of Science

  26. Diversity of the mammalian septins: Localization, interaction and functions

    M Kinoshita, M Valencik, T Haraguchi, Y Hiraoka, Pringle, JR, M Noda

    MOLECULAR BIOLOGY OF THE CELL   Vol. 9   page: 258A - 258A   1998.11

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  27. Mammalian septins: Involvement in cytokinesis, actin-based structures and neurodegenerative diseases

    M Kinoshita, A Kinoshita, T Haraguchi, Y Hiraoka, S Kumar, M Noda

    MOLECULAR BIOLOGY OF THE CELL   Vol. 8   page: 2145 - 2145   1997.11

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  28. Nedd5, a mammalian septin, is implicated in cytokinesis, stress fiber and focal adhesion

    M Kinoshita, T Haraguchi, Y Hiraoka, A Mizoguchi, C Ide, S Kumar, M Noda

    MOLECULAR BIOLOGY OF THE CELL   Vol. 7   page: 3237 - 3237   1996.12

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    Web of Science

  29. 脳虚血後の神経細胞死におけるシステインプロテアーゼcalpainとNedd2の関与

    木下専, 冨本秀和, 木下彩栄, KUMARSharad, 野田亮

    日本分子生物学会年会プログラム・講演要旨集   Vol. 19   1996.8

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Presentations 28

  1. 記憶の長期化に寄与するスパイン内小胞体の役割 Invited

    上田(石原)奈津実, 木下 専

    第19回神経科学研究会  2022.1.22  神経科学研究会

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    Event date: 2022.1

    Language:Japanese   Presentation type:Oral presentation (invited, special)  

    Venue:東京/オンライン   Country:Japan  

  2. Modification of physical properties and reconstruction of postsynaptic density by liquid-liquid phase separation Invited International conference

    Tomohisa Hosokawa, Pin-Wu Liu, Makoto Kinoshita

    2021.12.1 

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    Event date: 2021.12

    Language:English   Presentation type:Symposium, workshop panel (nominated)  

    Country:Japan  

  3. Activity-triggered extension of endoplasmic reticulum into dendritic spines as a synaptic basis of memory consolidation Invited International conference

    Makoto Kinoshita

    EMBO Workshop Molecular and cell biology of septins  2021.9.13  EMBO

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    Event date: 2021.9

    Language:English   Presentation type:Oral presentation (invited, special)  

    Venue:Berlin   Country:Germany  

  4. Calcium-induced persistent formation of protein condensate on Post-Synaptic Density by liquid-liquid phase separation Invited International conference

    Tomohisa Hosokawa, Pin-Wu Liu, Makoto Kinoshita, Yasunori Hayashi

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    Event date: 2021.7

    Language:English   Presentation type:Symposium, workshop panel (nominated)  

    Country:Japan  

  5. 液-液相分離によるシナプス後膜肥厚のカルシウム依存的かつ可塑的な蛋白質集合体の形成 International conference

    細川 智永、劉 品吾、木下 専、林 康紀

    第44回日本神経科学大会  2021.7.29  日本神経科学会

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    Event date: 2021.7

    Language:English   Presentation type:Symposium, workshop panel (nominated)  

    Venue:神戸   Country:Japan  

  6. 記憶固定化および長期記憶保持に寄与するポストシナプスのアクトミオシンとセプチンの協調 International conference

    上田(石原) 奈津実、深澤 有吾、高雄 啓三、見学 美根子、宮川 剛、井ノ口 馨、尾藤 晴彦、木下 専

    第44回日本神経科学大会  2021.7.30  日本神経科学会

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    Event date: 2021.7

    Language:English   Presentation type:Oral presentation (general)  

    Venue:神戸   Country:Japan  

  7. 記憶固定化のシナプス基盤:神経活動およびセプチン依存的なスパインへの小胞体伸展 International conference

    木下 専、上田-石原 奈津実、深澤 有吾、見学 美根子、高雄 啓三、井ノ口 馨、宮川 剛、尾藤 晴彦

    第43回日本神経科学大会  2020.7.31  日本神経科学学会

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    Event date: 2020.7 - 2020.8

    Language:English   Presentation type:Oral presentation (general)  

    Venue:神戸/オンライン   Country:Japan  

  8. Activity- and SEPT3-dependent ER entry into dendritic spines as a synaptic basis of memory consolidataion Invited

    Ageta-Ishihara N, Kosaka Y, Fukazawa Y, Takao K, Miyakawa T, Bito H, Inokuchi K, Kinoshita M

    Outcome Presentation Meeting FY2019. Platforms for Advanced Technologies and Research Resources  2020.2.4 

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    Event date: 2020.2

    Language:English   Presentation type:Poster presentation  

    Country:Japan  

  9. セプチン細胞骨格サブユニットSEPT3はスパインへの小胞体侵入を介して記憶保持に寄与する Invited

    木下 専

    第17回神経科学研究会  2019.11.23  大日本住友製薬

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    Event date: 2019.11

    Language:Japanese   Presentation type:Symposium, workshop panel (nominated)  

    Venue:東京   Country:Japan  

  10. ssTEM mapping of SEPT3 reveals its association with ER extending into active spines Invited International conference

    Fujiwara R, Kinoshita M

    The 7th Neural Circuits Joint Workshop  2019.10.17 

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    Event date: 2019.10

    Language:English   Presentation type:Symposium, workshop panel (public)  

    Country:Japan  

  11. Activity- and SEPT3-dependent ER entry into dendritic spines underlies long-term memory Invited International conference

    Asami Y, Kinoshita M

    The 7th Neural Circuits Joint Workshop  2019.10.17 

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    Event date: 2019.10

    Language:English   Presentation type:Symposium, workshop panel (public)  

    Country:Japan  

  12. Septin-dependent entry of smooth endoplasmic reticulum into dendritic spines as a synaptic basis of persistent memory Invited International conference

    Ageta-Ishihara N, Fukazawa Y, Takao K, Miyakawa T, Bito H, Inokuchi K, Kinoshita M

    OIST Mini-Symposium: The 16th International Membrane Research Forum  2019.3.18  OIST

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    Event date: 2019.3

    Language:English   Presentation type:Symposium, workshop panel (nominated)  

    Venue:OIST   Country:Japan  

  13. Requirement of a septin subunit SEPT3 in the dentate gyrus granule cells for spine maturation and spatial discrimination Invited

    2018.12.4 

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    Event date: 2018.12

    Language:Japanese   Presentation type:Symposium, workshop panel (nominated)  

    Country:Japan  

  14. Requirement of a septin subunit SEPT3 in the dentate gyrus granule cells for dendritic spine maturation and spatial discrimination International conference

    Sakakibara K, Fukumasu N, Ageta-Ishihara N, Fukazawa Y, Takao K, Miyakawa T, Bito H, Kinoshita M

    The 6th Neural Circuits Joint Workshop  2018.10.5  NSI

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    Event date: 2018.10

    Language:English   Presentation type:Poster presentation  

    Country:Japan  

  15. Unexpected motor and histopathological phenotype found in mice that lack SEPT7 in the cerebellum International conference

    Ageta-Ishihara N, Kurita H, Yamazaki M, Abe M, Fukazawa Y, Sakimura K, Kinoshita M

    The 6th Neural Circuits Joint Workshop  2018.10.5  NSI

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    Event date: 2018.10

    Language:English   Presentation type:Poster presentation  

    Country:Japan  

  16. A postsynaptic dysregulation in hippocampal granule cells that underlies spatial discrimination defect Invited International conference

    Ageta-Ishihara N, Fukazawa Y, Takao K, Miyakawa T, Bito H, Inokuchi K, Kinoshita M

    The Joint Congress of the 40th Annual Meeting of Japanese Society of Biological Psychiatry and the 61st Annual Meeting of the Japanese Society for Neurochemistry  2018.9.7  The Japanese Society of Biological Psychiatry, The Japanese Society for Neurochemistry

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    Event date: 2018.9

    Language:English   Presentation type:Symposium, workshop panel (public)  

    Country:Japan  

  17. An activity-regulated septin subunit SEPT3 is required for the entry of sER into dendritic spines and spatial discrimination Invited International conference

    2018.6.28 

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    Event date: 2018.6

    Language:English   Presentation type:Symposium, workshop panel (nominated)  

    Country:United States  

  18. Entorhinal cortex-hippocampal dentate gyrus synapse anomaly underlying spatial discrimination defects in Sept3-null mice Invited

    Makoto Kinoshita

    2018.5.10 

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    Event date: 2018.5

    Language:English   Presentation type:Symposium, workshop panel (nominated)  

    Venue:OIST   Country:Japan  

  19. CDC42EP4, a major septin-binding protein in astroglia, is required for glutamatergic tripartite synapse configuration Invited International conference

    Kinoshita M

    EMBO Workshop on Molecular and Cell Biology of Septins  2017.10.13  EMBO

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    Event date: 2017.10

    Language:English   Presentation type:Symposium, workshop panel (nominated)  

    Venue:Freie Universität Berlin, Berlin, Germany  

  20. セプチン欠損マウスを用いたシナプス制御機構の探索 Invited

    木下 専

    第1回先端領域融合医学研究会  2017.10.7  先端領域融合医学研究会

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    Event date: 2017.10

    Language:Japanese   Presentation type:Symposium, workshop panel (public)  

    Venue:神戸医療産業都市推進機構 先端医療研究センター   Country:Japan  

  21. セプチン細胞骨格サブユニットSEPT3の欠損による空間弁別障害 Invited

    木下 専

    第15回神経科学研究会  2017.10.7  大日本住友製薬

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    Event date: 2017.10

    Language:Japanese   Presentation type:Symposium, workshop panel (nominated)  

    Venue:東京   Country:Japan  

  22. セプチン細胞骨格系の生理機能の探索 Invited

    木下 専

    JSTさきがけ「生体分子の形と機能」10年間の展開  2017.9.19  大阪市立大学理学研究科宮田真教授

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    Event date: 2017.9

    Language:Japanese   Presentation type:Symposium, workshop panel (nominated)  

    Venue:大阪市立大学   Country:Japan  

  23. Molecular dynamics driving neuronal morphogenesis Invited International coauthorship International conference

    Makoto Kinoshita

    2016.7.22 

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    Event date: 2016.7

    Language:English   Presentation type:Symposium, workshop panel (nominated)  

    Country:Japan  

  24. トランスポーター(EAATs/VGLUTs)によるグルタミン酸分布の時空間制御と精神・神経疾患 Invited International conference

    木下 専

    第37回日本神経科学大会  2014.9.13  世話人:木下 専、田中 光一

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    Event date: 2014.9

    Language:English   Presentation type:Symposium, workshop panel (nominated)  

    Venue:横浜   Country:Japan  

  25. セプチン変異マウスを用いた未知の空間学習・記憶メカニズムの探索 Invited International conference

    上田 ( 石原 ) 奈津実、木下 専

    第36回日本分子生物学会年会  2013.12.4  世話人:木下 専、喜多村 和郎

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    Event date: 2013.12

    Language:Japanese   Presentation type:Symposium, workshop panel (public)  

    Venue:神戸   Country:Japan  

    記憶―動的環境情報を神経ネットワークに書き込む分子システム基盤―への多階層アプローチ

  26. Three dimensional distribution and stability of septin clusters in the mammalian brain Invited International coauthorship International conference

    BMB2007  2007.12.15 

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    Event date: 2007.12

    Language:Japanese   Presentation type:Symposium, workshop panel (nominated)  

    Country:Japan  

  27. Sept4, a component of presynaptic scaffold and Lewy bodies, is required for the suppression of alpha-synuclein neurotoxicity International coauthorship International conference

    Makoto Kinoshita

    Neuro 2007  2007.9.11 

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    Event date: 2007.9

    Language:English   Presentation type:Oral presentation (general)  

    Country:Japan  

  28. Unexpected motor and histopathological phenotype found in cerebellum-selective Sept7 knockout mice Invited

    Kinoshita M

    The 8th Brain Research Institute International Symposium“Innovative progress of neuroscientific research through the use of advanced animal models”  2018.2.10  Brain Research Institute, Niigata University

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    Language:English   Presentation type:Poster presentation  

    Country:Japan  

▼display all

Works 7

  1. 遺伝子改変マウスC57BL/6-Sept7<tm1Ksh>

  2. 遺伝子改変マウスC57BL/6-Cdc42ep4<tm1.1Ksh>

    木下 専

  3. 遺伝子改変マウスC57BL/6-Tg(Prnp-Sirt1,Cryaa-EGFP)1Ksh

    木下 専

  4. 遺伝子改変マウスC57BL/6-Cdc42ep4<tm1Ksh>

    木下 専

  5. 遺伝子改変マウスC57BL/6J-Tg(Prnp-Sept4)1Ksh

    木下 専

  6. 遺伝子改変マウス C.129X1(B6)-Sept4<tm1Ksh>

    木下 専

  7. 遺伝子改変マウス B6.129X1-Sept4<tm1Ksh>

    木下 専

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Research Project for Joint Research, Competitive Funding, etc. 15

  1. 細胞内に侵入した病原微生物を捕捉する新たな感染防御メカニズムの解析 International coauthorship

    2012.4 - 2013.3

    旭硝子財団 研究奨励 

    木下 専

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    Authorship:Principal investigator  Grant type:Competitive

    Grant amount:\2000000

  2. トランスジェニック動物マーカーの開発と応用

    2009.4 - 2010.3

    地域イノベーション創出支援事業 重点地域研究開発推進プログラム シーズ発掘試験A 

    木下 専

      More details

    Grant type:Competitive

  3. Travel Grant for Janelia Conference "Structure and Function of Septins" (at Ashburn, VA, USA) International coauthorship

    2009.3

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    Authorship:Principal investigator  Grant type:Competitive

    Grant amount:\154910

  4. 動物モデルと臨床検体を用いた肺線維症病態メカニズムの解析

    2008.4 - 2009.3

    病態代謝研究会 研究助成金 

    木下 専

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    Authorship:Principal investigator 

    Grant amount:\1000000

  5. パーキンソン病遺伝子ネットワーク解明と新規治療戦略

    Grant number:JPMJCR0734  2007.10 - 2013.3

    戦略的創造研究推進事業 CREST「精神・神経疾患の分子病態理解に基づく診断・治療へ向けた新技術の創出」 

    高橋良輔

      More details

    Authorship:Coinvestigator(s)  Grant type:Competitive

    パーキンソン病責任蛋白質(変異型alpha-synuclein)と関連因子(変異型tau, septin欠損)に関する多因子改変マウスを樹立して分子病態を解析するとともに分子標的治療シーズを探索する。

  6. アルツハイマー病(tauopathy)モデルマウスを用いた脆弱性因子の検討

    2007.4 - 2008.3

    武田科学振興財団 医学系研究奨励  

    木下 専

      More details

    Authorship:Principal investigator  Grant type:Competitive

    Grant amount:\2000000

  7. 凍結割断レプリカ多重標識法によるセプチン細胞骨格および関連分子の細胞膜直下における高分解能局在解析

    2005.4 - 2006.3

    自然科学研究機構 生理学研究所 共同利用研究 

    木下 専

      More details

    Authorship:Principal investigator  Grant type:Competitive

    Grant amount:\200000

  8. 早発性神経変性性障害のトランスジェニック動物モデルの開発と応用

    2005.4 - 2006.3

    研究成果展開事業 地域イノベーション創出総合支援事業 重点地域研究開発推進プログラム シーズ発掘試験 

    木下 専

      More details

    Authorship:Principal investigator 

  9. American Society for Cell Biology (Washington D.C., USA) 参加旅費

    2004.12

    金原一郎記念医学医療振興財団 第18回研究交流助成金 

    木下 専

      More details

    Authorship:Principal investigator 

    Grant amount:\180000

  10. 肝脂質代謝におけるセプチン系の解析

    2004.4 - 2005.3

    成人病の病因・病態の解明に関する研究会 研究助成金 

    木下 専

      More details

    Authorship:Principal investigator 

    Grant amount:\400000

  11. Traveling fee for Gordon Research Conference at Oxford University

    2004.4

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    Authorship:Principal investigator  Grant type:Competitive

    Grant amount:\150000

  12. 極低温電子線断層法によるセプチン系超分子構造体の解析

    Grant number:JPMJPR0362  2003.10 - 2007.3

    戦略的創造研究推進事業 個人型研究(さきがけ) 生体分子の形と機能 

    木下 専

      More details

    Authorship:Principal investigator  Grant type:Competitive

    Grant amount:\41700000

  13. 2つのパーキンソン病関連蛋白と脂質間相互作用の分子病理学的解析

    2003.4 - 2004.3

    武田科学振興財団 医学系研究奨励 

    木下 専

      More details

    Authorship:Principal investigator 

    Grant amount:\2000000

  14. Biochemical and ultrastructural analysis of the mammalian septins International coauthorship

    2000.1 - 2001.12

    Human Frontier Science Program Long-term fellowship 

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    Authorship:Principal investigator  Grant type:Competitive

    Grant amount:\5000000

  15. アポトーシス誘導活性を有する新規システインプロテアーゼNedd2と相互作用する分子の単離と解析

    1996.4 - 1997.3

    稲盛財団 研究助成金 

    木下 専

      More details

    Authorship:Principal investigator  Grant type:Competitive

    Grant amount:\1000000 ( Direct Cost: \1000000 )

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KAKENHI (Grants-in-Aid for Scientific Research) 36

  1. Investigation on a novel synaptic mechanism underlying memory consolidation

    Grant number:21K19427  2021.7 - 2023.3

    Grants-in-Aid for Scientific Research  Grant-in-Aid for Challenging Research (Exploratory)

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    Authorship:Principal investigator  Grant type:Competitive

    Grant amount:\6500000 ( Direct Cost: \5000000 、 Indirect Cost:\1500000 )

  2. 2相性インスリン極性分泌機構とその破綻による糖尿病発症

    Grant number:21H02431  2021.4 - 2024.3

    日本学術振興会  科学研究費助成事業  基盤研究(B)

    今泉 美佳, 青柳 共太, 木下 専, 大塚 稔久, 安田 和基

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    Authorship:Coinvestigator(s)  Grant type:Competitive

    Grant amount:\17290000 ( Direct Cost: \13300000 )

    膵β細胞からのインスリン分泌は2相性であり、毛細血管方向へ極性分泌されるがその分子機構は未だ不明な点が多く、インスリン分泌不全を呈する2型糖尿病の病態を明らかにするためにも解明が急がれている。本研究ではβ細胞の毛細血管に面した細胞膜領域に偏って局在しているアクティブゾーンタンパク質群とセプチン重合体に着目し、2相性インスリン極性分泌機構の全容を解明することを目的とする。

  3. シナプス可塑性の基盤となる液-液相分離制御機構の解明

    Grant number:21F21384  2021.11 - 2024.3

    日本学術振興会  特別研究員奨励費  特別研究員奨励費

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    Authorship:Collaborating Investigator(s) (not designated on Grant-in-Aid)  Grant type:Competitive

    Grant amount:\2300000 ( Direct Cost: \2300000 )

  4. シナプス活動で遊離したセプチンによるPSDと小胞体の再編が記憶固定化に寄与する

    Grant number:21H02579  2021.4 - 2024.3

    文部科学省  科学研究費補助金  基盤研究(B)

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    Authorship:Principal investigator  Grant type:Competitive

    Grant amount:\17420000 ( Direct Cost: \13400000 、 Indirect Cost:\4020000 )

  5. Exploration of synaptic mechanism underlying spatial cognitive defect of septin-null mice

    Grant number:18H02525  2018.4 - 2021.3

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    Authorship:Principal investigator 

    Grant amount:\17420000 ( Direct Cost: \13400000 、 Indirect Cost:\4020000 )

  6. 嗅内皮質-海馬歯状回シナプス機能不全による空間弁別障害と適応的代償機構の解析

    Grant number:17H05564  2017.4 - 2019.3

    科学研究費補助金(新学術領域研究(研究領域提案型))   新学術領域研究(研究領域提案型)

    木下 専

      More details

    Authorship:Principal investigator 

    Grant amount:\7020000 ( Direct Cost: \5400000 、 Indirect Cost:\1620000 )

    セプチン細胞骨格系を構成するサブユニットSEPT1-14のうち、SEPT3は脳特異的かつニューロン選択的に発現するが、生理機能は不明である。Sept3欠損マウスは空間文脈課題で長期記憶障害を呈する。責任領域(海馬歯状回;DG)の主要ニューロン(顆粒細胞)の樹状突起棘(スパイン)の体積は正常であるが滑面小胞体(ER)含有率が少なく、この異常は初代培養顆粒細胞のSEPT3枯渇によって再現された。スパイン内ERの役割として、Ca2+シグナル伝達や膜輸送を介してシナプス伝達ないし長期増強への寄与が推測されている。Sept3欠損マウスでER含有スパインが減少するメカニズムとして、①顆粒細胞のER生合成の減少、②スパインへの侵入率の減少・退出率の増加、が考えられる。本研究では、①の検証を目的として、透過型電子顕微鏡連続切片像3次元再構築法(serial section Transmission Electron Microscopy; ssTEM法)により、野生型およびSept3欠損マウスの樹状突起内ER体積を比較した。その結果、Sept3欠損マウスの長期記憶障害の責任領域であるDG領域において、ER含有スパインの割合が有意に低いことを再確認した。一方、樹状突起基幹部内のER体積、基部にER集積部を持つスパインの割合には野生型との間に有意差を認めなかったことから、上記①(ER生合成ないし樹状突起基幹部内のERネットワーク減少に起因する可能性)は否定された。樹状突起基幹部内で局所的に形成されるER集積部の生理機能は不明である。本研究で、スパイン基部におけるER集積とスパイン内ERに強い関連を認めたことから「ER集積部は、スパイン内へのER侵入を効率化する足場としてシナプス入力依存的に形成される」いう新たな仮説を立てた。
    平成30年度が最終年度であるため、記入しない。
    平成30年度が最終年度であるため、記入しない。

  7. 3D morphometry of synapses with focused ion beam-scanning electron microscopy

    Grant number:16K14767  2016.4 - 2018.3

    KINOSHITA MAKOTO

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    Authorship:Principal investigator 

    Grant amount:\3640000 ( Direct Cost: \2800000 、 Indirect Cost:\840000 )

    In this study we acquired serial 2D images from conventionally prepared tissue samples from wild type and genetically engineered mouse brain and compared 3D reconstructed images by using conventional ssTEM and FIB-SEM (in collaboration with the Fukazawa lab at Fukui Univ.). FIB-SEM, but not ssTEM, found a unique anomaly in synapse morphology, mainly due to the thickness along Z-axis (5nm vs. 50nm), whereas ssTEM could cover wider volume than FIB-SEM. Comparison of the two methods with immunogold-labeled samples is under investigation.

  8. 放射状グリア突起内CDC42EP4セプチン複合体の生理機能と凝集体残留機構の解析

    Grant number:16H01334  2016.4 - 2018.3

    科学研究費補助金(新学術領域研究(研究領域提案型))   新学術領域研究(研究領域提案型)

    木下 専

      More details

    Authorship:Principal investigator 

    Grant amount:\8840000 ( Direct Cost: \6800000 、 Indirect Cost:\2040000 )

    小脳バーグマングリア選択的に発現するCDC42EP4に着目して遺伝子欠損マウス系統を樹立し、以下の所見を得た。①平行線維-プルキンエ細胞間のグルタミン酸作動性シナプスを被覆するバーグマングリア突起において、CDC42EP4とセプチン線維はシナプスに面する細胞膜直下に集積する。②両者は複合体を形成し、ミオシン-10やグルタミン酸トランスポーターGLAST/EAAT1とも相互作用する。③CDC42EP4欠損によってこの相互作用が減弱し、GLASTはシナプスから遠ざかる方向に脱局在する。④電気生理学的には上記シナプスのEPSPが軽度に遷延するのみであるが、グルタミン酸トランスポーター阻害剤DL-TBOAの添加に過敏に反応し、後シナプス膜電位が顕著に上昇する。⑤運動学習障害は軽度であるが、野生型では無効な低濃度のDL-TBOAの投与によって重篤化する。以上から、CDC42EP4-セプチン複合体はGLASTをシナプス近傍に集積させる足場ないし拡散障壁としてグルタミン酸クリアランスに寄与することが示唆された(Nat Commun 2015)。グルタミン酸クリアランスを遅延させる形態異常の合併の可能性を精査したところ、CDC42EP4欠損マウスではBG突起先端がシナプス間隙から後退する一方、前後シナプス膜の接着領域が辺縁部でのみ拡大していた。N-cadherinや非筋型ミオシン重鎖ⅡB/MYH-10などニューロン側の接着関連分子の増加や、最近報告されたGLAST欠損マウス(宮崎らPNAS 2017)との類似性から、この異常は過剰なグルタミン酸に対するニューロン側の2次的反応と推測される。グルタミン酸滞留に起因するこのようなシナプス再編成は、細胞外死腔を拡大することでグルタミン酸クリアランスをさらに遅延させ、悪循環をもたらす可能性がある(Neurochem Int 2018)。
    29年度が最終年度であるため、記入しない。
    29年度が最終年度であるため、記入しない。

  9. Molecular network and physiological function of glutamatergic tripartite synapse in the cerebellum

    Grant number:15H04274  2015.4 - 2018.3

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research  Grant-in-Aid for Scientific Research (B)

    KINOSHITA MAKOTO, ISHIHARA NATSUMI

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    Authorship:Principal investigator 

    Grant amount:\18070000 ( Direct Cost: \13900000 、 Indirect Cost:\4170000 )

    The perforant path (pp)-granule cell (GC) synapse is poorly characterized at molecular/subcellular level. Here we show that mice that lack a pan-neuronal subunit of the septin cytoskeleton, pass hippocampus-dependent tasks. However, they consistently underperform in specific tasks that require discrimination among distinct spatial contexts. Hippocampal dentate gyrus (DG) neuron-selective supplementation of the septin subunit restores their performance, while the local depletion recapitulates the defects in wild-type mice. EM morphometry of three major hippocampal subregions shows normal synapse density, PSD area, and spine volume, but reveals a significant scarcity of spine apparatus that is most severe in the pp-GC synapse. Live imaging of septin-depleted primary cultured DG neurons reveals low frequency in ER entry from dendritic shafts into spines. These and other findings indicate a septin-mediated postsynaptic mechanism required in pp-GC synapses for spatial pattern separation.

  10. 嗅内皮質-海馬歯状回シナプス分子欠乏による空間失認と適応的代償機構の解析

    Grant number:15H01429  2015.4 - 2017.3

    文部科学省  科学研究費補助金(新学術領域研究(研究領域提案型))  新学術領域研究(研究領域提案型)

    木下 専

      More details

    Authorship:Principal investigator  Grant type:Competitive

    Grant amount:\6890000 ( Direct Cost: \5300000 、 Indirect Cost:\1590000 )

    空間認知・記憶を担う海馬神経回路のうち、嗅内皮質(貫通線維)から海馬歯状回(顆粒細胞)への投射 (pp-GC)は発火閾値が高く、sparse codingによるパターン分離(空間文脈の弁別)に重要とされている。このユニークなグルタミン酸作動性シナプスの樹状突起棘(スパイン)には重合性GTPaseセプチンの複数のサブユニットが局在するが、生理的意義は不明である。我々は、海馬歯状回顆粒細胞に高発現する機能未知のセプチン細胞骨格サブユニットの1つに着目し、その遺伝子欠損マウスの神経機能異常を系統的行動解析でスクリーニングしたところ、空間定位・記憶を含めて行動様式がほぼ正常であるにもかかわらず、異なる空間文脈を混同するという、限局的な空間認知・弁別障害を見出した。そこで、1)野生型マウス歯状回へのRNAi注入実験により、顆粒細胞のセプチンが空間弁別に必要であること、2) レスキュー実験により、欠損マウスの顆粒細胞でのセプチン発現が空間弁別に十分であることを示し、空間弁別障害の主要な責任領域がpp-GCシナプスであることを確定した。引き続き、セプチンの欠乏ないし欠損からシナプス機能障害に至るメカニズムの解明を目指して、蛍光ライブイメージング、神経薬理学的操作、プロテオミクスを組み合わせた探索を進めている。行動>海馬神経回路>シナプス活動>シナプス微細構造>シナプス分子の階層での異常を詳細に精査することを通じて、認知症や統合失調症などの精神・神経疾患の部分症状として頻度の高い空間見当識障害の病態理解につながることも期待される。
    28年度が最終年度であるため、記入しない。
    28年度が最終年度であるため、記入しない。

  11. 免疫電子顕微鏡(凍結割断および3次元)を用いたシナプスとグリアの微細形態異常解析

    Grant number:25116514  2013.4 - 2015.3

    文部科学省  科学研究費補助金(新学術領域研究(研究領域提案型))  新学術領域研究(研究領域提案型)

    木下 専

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    Authorship:Principal investigator  Grant type:Competitive

    Grant amount:\7020000 ( Direct Cost: \5400000 、 Indirect Cost:\1620000 )

    前・後のシナプス膜上には神経伝達物質の開口放出装置/受容体/トランスポーターやイオンチャネルなどシナプス伝達の最前線の分子群が集積する一方、シナプス膜直下の細胞骨格ないし足場蛋白質もシナプス機能に不可欠である。例えば、興奮性シナプス後膜の脱分極に伴うアクチン重合は樹状突起棘(スパイン)容積を増加させ、シナプス伝達の長期増強(LTP)持続の支持基盤となる。シナプス膜直下の細胞骨格/足場の構成成分であるセプチン(SEPT1-14)は精神・神経疾患に密接に関連する重合性GTP結合蛋白質である。統合失調症に限っても、大規模GWASにおいてSEPT3が責任遺伝子候補とされ(Schizophrenia Working Group, Nature 2014)、死後脳プロテオームの大規模解析でも多数例でSEPT5、SEPT6、SEPT11の過剰蓄積が報告された(PenningtonらMol Psychiatry 2008)。本研究ではスパイン内に局在するSEPT3の欠損が空間認知・弁別機能障害をもたらすことを新たに見出した。原因として、回路レベルでは貫通線維―海馬歯状回顆粒細胞(pp-GC)間グルタミン酸作動性シナプスの伝達障害を、シナプスレベルではSDS処理凍結割断レプリカ免疫標識電子顕微鏡法によってGluA1の欠乏を、連続切片像3D再構築(ssTEM)電子顕微鏡法ではスパイン内小胞体の貧弱化を認めた。引き続き関連データの取得を行い、研究を推進する予定である。
    26年度が最終年度であるため、記入しない。
    26年度が最終年度であるため、記入しない。

  12. Exploration of physiological functions of mammalian septins with genetically engineered mouse lines

    Grant number:23370084  2011.4 - 2014.3

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research  Grant-in-Aid for Scientific Research (B)

    KINOSHITA Makoto

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    Authorship:Principal investigator  Grant type:Competitive

    Grant amount:\20280000 ( Direct Cost: \15600000 、 Indirect Cost:\4680000 )

    Septins are evolutionarily conserved polymerizing GTPases like the major cytoskeletal nucleotide-binding proteins of tubulins and actins, however, their physiological functions in metazoans are largely unknown. To address this: 1) We have established a line of conditional SEPT7 knockout mice and uncovered that septins promote dendrite and axon development by negatively regulating microtubule stability via HDAC6-mediated deacetylation. 2) We established prion-promoter-driven SEPT4 transgenic mice and uncovered that chronic overload of SEPT4, a parkin substrate that aggregates in Parkinson's disease, causes behavioral alterations but not neurodegeneration in mice. These and other findings, in addition to mouse lines and a set of reagents created in this study, will help understand cellular/molecular mechanisms of septin functions in relation to other cytoskeletal systems in physiologically relevant contexts.

  13. 前シナプス膜とグリア膜直下のセプチン細胞骨格の破綻を伴う精神・神経疾患病態の解析

    Grant number:23110531  2011.4 - 2013.3

    文部科学省  科学研究費補助金(新学術領域研究(研究領域提案型))  新学術領域研究(研究領域提案型)

    木下 専

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    Authorship:Principal investigator  Grant type:Competitive

    Grant amount:\5590000 ( Direct Cost: \4300000 、 Indirect Cost:\1290000 )

    重合性GTPaseであるセプチン(SEPT1-14)はニューロンやグリアの細胞骨格の構成要素である。優性変異型SEPT9による家族性ニューロパシー(KuhlenbaumerらNat Genet 2005)、統合失調症や双極性障害におけるSEPT5/6/11の増加(PenningtonらMol Psychiatry 2008)、ParkinによるSEPT4とSEPT5のユビキチン化などが報告されてきたが(ZhangらPNAS 2001など)、いずれも病態生理学的意義は不明である。シナプス病態班員としての研究期間を通じて以下の進捗があった。
    パーキンソン病の病態モデルとして「Parkinの変異で分解を免れたSEPT4などの基質の蓄積が神経を障害する」仮説が優勢である。そこで脳選択的SEPT4過剰発現マウスを作製して検証したが、軽度の自発活動量低下傾向以外に行動学的異常はなく、凝集体も含めた病理変化もなく、上記仮説の反証となった(in revision)。
    黒質-線条体ドーパミン伝達が減弱するSEPT4欠損マウス(猪原、木下らNeuron 2007)より重篤な病態を期待して、必須サブユニットを脳の一部で欠損するマウスを作製したところ、パーキンソン病末期に出現するアミロイド小体に類似した微小な凝集体が多発した。凝集体は幼若マウスにおいて既に多発しており、ほぼ全てのセプチンを含み、細胞質成分を巻き込んだ独特の形状であった。中間径線維や隔離膜で包囲されていなかったことから、アグリソームとして封入されていないことがわかった。多数の微小凝集体が残留する原因として、凝集蛋白質の輸送、アグリソームへの集積、オートファジーによる分解系からエスケープしている可能性を考え、これらパスウェイの既知分子を解析したところ、特定の蛋白質が著減していることがわかった(in preparation)。
    24年度が最終年度であるため、記入しない。
    24年度が最終年度であるため、記入しない。

  14. Exploration for the mechanism of exercise-induced neural activation and protection

    Grant number:23650191  2011

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

    KINOSHITA Makoto

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    Authorship:Principal investigator  Grant type:Competitive

    Grant amount:\4030000 ( Direct Cost: \3100000 、 Indirect Cost:\930000 )

    By using a line of transgenic mouse line in which GFP expression is driven by a neural activity-dependent promoter, we mapped brain areas where neural activity is upregulated after exercise. In mice underwent voluntary exercise on running wheels, GFP fluorescence became significantly more intense than the control mice in the following areas : hippocampus(medial CA1 area, pyramidal layer, medial dentate gyrus), entorhinal cortex, somatosensory area(S1). Since a subset of the dentate gyrus neurons were positive in GFP and phosphorylated TrkB(indicating activation of the major BDNF receptor) we conducted proteomic analysis of this area using comparative shotgun LC-MS/MS method. The data indicated that a few cytoskeletal proteins, adaptor proteins, metabolic enzymes(whose identities will be disclosed upon publication of the study) were significantly upregulated in activity-dependent manner. After validation with immunoblot and immunohistochemistry, we focused on one candidate protein whose cognate gene knockout mouse line had been deposited in a public repository. We have already completed backcrossing of the line to the C57BL/6N background. Using mice with or without the candidate gene, we are preparing for comparative analysis on exercise-mediated biochemical, metabolic, behavioral changes and susceptibility against oxidataive and/or proteotoxic stresses.

  15. 自己組織性蛋白質と脂質二重膜のダイナミックな相互作用の直接観察と動力学的解析

    Grant number:21015010  2009 - 2011

    科学研究費補助金(特定領域研究)  特定領域研究

    滝口金吾

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    Authorship:Coinvestigator(s) 

    細胞や細胞内小器官は、脂質二重膜を基本構造に持つ生体膜によって区画されその機能を維持している。生体膜の形態調節や変形、移動運搬は膜と相互作用する細胞骨格線維をはじめとする蛋白質によって制御されると考えられているが、その詳しい機構はまだ分かっていない。本研究では、神経シナプスのスパインや細胞分裂時の分裂溝に存在するセプチンを膜小胞に添加すると激しい膜突起形成が生じることを明らかにした。
    以上のように新たに発見されたセプチンの様な蛋白質による膜突起形成機構を理解するために、線維形成といった異方性を持つ蛋白質の膜への結合の効果を組込んだ膜弾性モデルを使用して、膜小胞の形状変化過程を解析した。膜の曲げエネルギーが、結合した蛋白質の密度およびその配向に応じて決定されるモデルから計算された結果は、膜周囲の溶液中に充分量の蛋白質が存在すると、突起が小胞から伸長することを示し、実験結果を良く再現した。
    ところで、細胞分裂時に膜変形力を発生する細胞膜直下の蛋白質ネットワークのうち、これまではアニリンが膜に直接会合すると考えられていた。この仮説を検証するために、人工リン脂質膜と組み換え蛋白質を用いた実験を行った結果、セプチンとは異なりアニリンは直接膜とは相互作用しないことが明らかとなり、細胞膜-セプチン-アニリンという分子構築が示唆された。アニリンは細胞骨格アクチンおよび分子モーターのミオシンへの連結を担うと考えられる。

  16. Mechanism of cortical septin heteropolymerization/dissociation

    Grant number:20370077  2008 - 2010

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research  Grant-in-Aid for Scientific Research (B)

    KINOSHITA Makoto

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    Authorship:Principal investigator  Grant type:Competitive

    Grant amount:\20670000 ( Direct Cost: \15900000 、 Indirect Cost:\4770000 )

    Despite the cortical localization of septin heteropolymers in vivo and their direct interaction with phospholipid membranes in vitro, their behavior and roles remain elusive. In this project, we characterized the major cortical septin assembly found in mammalian tissue culture cells by fluorescence recovery after photobleaching (FRAP) analysis. GFP-tagged septin subunits exhibited slower turnover compared with other cortical proteins analyzed. Perturbation of actin turnover retarded the cortical septin turnover, while septin depletion by RNAi did not affect cortical actin turnover. These phenomena are interpreted by septins' selective association with a subset of actin-based membrane skeleton, as revealed by quick-freeze deep-etch immunoelectron microscopy. We applied the assay system to test septins' presumptive scaffold function on their physiological binding partners. Septin filament destabilization by RNAi-mediated subunit depletion facilitated the turnover of GLAST, while a septin-stabilizing drug forchlorfenuron restrained more GLAST in the unexchangeable fraction. Thus, cortical septin heteropolymers are components of the actin-based membrane skeleton, providing scaffolds for their interacting partners by impeding their lateral diffusion. We predict that diverse submembranous septin clusters found in vivo may serve as scaffolds or reserve pools for specific membrane-bound proteins.

  17. プルキンエ細胞およびバーグマングリア特異的Sept7欠損マウスの解析

    Grant number:20022022  2008 - 2009

    文部科学省  科学研究費補助金(特定領域研究)  特定領域研究

    木下 専

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    Authorship:Principal investigator  Grant type:Competitive

    Grant amount:\8000000 ( Direct Cost: \8000000 )

    Sept7(exon3)floxedマウスを作製し、L7Cre、S100b-CreおよびDAT-Creドライバーマウスと交配した。組み換えはPCRでは検出されるものの、組織染色ではSept7蛋白質欠損細胞は検出されなかったことから、両アレル同時には欠失しない程度に組み換え効率が低いと推測された。一方、Tリンパ球特異的なCd4-Creでは組み換えが起こって期待される形質が発現したことから(投稿準備中)、Sept7(exon3)floxedアレルの組み換え効率が分化状態ないし細胞系譜による影響を受けやすいことがわかった。対策として、1 hitでnullが得られるようdel/floxedマウスの作製を開始した。また、支援班の崎村教授グループの協力を得て(GluR2-Creマウスとの交配を試みるとともに、組み換え効率の改善を期待してSept7(exon4)floxedマウスを作製した。
    関連研究として、大脳皮質ニューロンにおけるSept7RNAiが軸索・樹状突起の伸長を阻害することを示した。これは線虫のセプチン変異体unc-59/-61で報告された神経突起伸長・分岐異常と符合する。このメカニズムが既知のセプチン機能(微小管、アクチン、tSNAREの機能修飾・安定化)で説明できるか否かを検証中である。また、低分子量G蛋白質Cdc42はエフェクター蛋白質ファミリーCdc42ep1-/1Borg1-5を介してセプチン系を制御する(Nat Cell Biol 2001)。このパスウェイの生理的意義を検証するため、バーグマングリアと海馬歯状回ニューロンで選択的に発現するCdc42ep/Borgに着目してノックアウトマウスを作製し、解析中である。
    以上、本研究で作製した研究リソースが、脳研究に貢献し、さらに、がん、不妊、肝線維化、感染防御、免疫系の研究にも役立つ得ることがわかった。

  18. 膜骨格系による膜輸送複合体の安定化機構

    Grant number:20056017  2008 - 2009

    日本学術振興会  科学研究費助成事業  特定領域研究

    木下 専

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    Authorship:Principal investigator  Grant type:Competitive

    Grant amount:\5000000 ( Direct Cost: \5000000 )

    (1) マウスゲノムに存在する13種類のセプチン遺伝子のうち成熟脳特異的に発現するSept4に着目して遺伝子破壊を行い、網羅的行動解析によってSept4欠損マウスのドパミン神経伝達が減弱していることを見出した。Sept4を含むセプチン複合体がドパミン・トランスポーター(DAT/Slc6A3)やtSNAREを含むトランスポートソームの構成成分であることなどから、シナプス前スカフォールドとしてのセプチン系の新たな役割が示された。さらに、パーキンソン病においてSept4が減少することがドパミン神経伝達を低下させ、病態を加速させる可能性も示唆された。セプチン系の破綻が微小管による膜蛋白質の輸送障害をもたらすことも、複雑な病態の背景にあるかもしれない。
    (2) マウス神経組織におけるセプチン分子の局在から機能に関する手がかりを得るため、免疫電子顕微鏡(immunogold法)連続切片像(70nm厚×20切片)からシナプスやグリア突起形状と金粒子の分布情報を抽出してコンピュータ上で3次元再構築した。これによりシナプス近傍やグリア突起における特定の細胞膜ドメインにおいてセプチンが集積する傾向が明らかになった。これら細胞膜直下のセプチン・クラスターの一部はGLAST/Slc1A3などの神経伝達物質輸送体やその他重要な膜蛋白質と共局在することから、DATでみられたようにトランスポートソームのスカフォールドを構成している可能性がある。また、一部は膜骨格として樹状突起棘(スパイン)やグリア突起形状の構造的支持に寄与する可能性がある。

  19. 自己組織性ポリペプチドと脂質二重膜相互作用の直接観察と動力学的解析

    Grant number:19031012  2007 - 2008

    日本学術振興会  科学研究費助成事業  特定領域研究

    瀧口 金吾, 木下 専, 梅田 民樹

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    セプチンは植物以外の全ての真核生物から見いだされるよく保存されている蛋白質である。セプチンが線維形成能を持ち、細胞の形態形成や運動などに必須なことから、セプチンは第4の細胞骨格と考えられている。セプチンの欠損や異常が、生物に致死を招き、神経変性、不妊症、腫瘍発生などの多くの疾患の発症進展に関与していることなど、その重要性が数多く示されてきたにも関わらず、セプチンの基本的な性質や機能はまだ不明であった。
    今までよく知られている細胞骨格とセプチンが異なるのは、リン脂質と結合して直接生体膜に作用できるところである。そこで本研究では、ブタやラットの脳から得たセプチンを含む画分や、昆虫細胞で発現させ単離してきたセプチンを、細胞とほぼ同じサイズを持つ巨大リポソームに作用させ、リポソーム膜に対してセプチンがどのような作用を及ぼすかを暗視野顕微鏡を用いてリアルタイム観察した。その結果セプチンに膜突起を誘導する強力な活性があることを世界で初めて明らかにし、膜突起の形成伸長過程を可視化することに成功した。暗視野顕微鏡でリポソームの変形を予め確認しておいた試料を電子顕微鏡で観察したところ、セプチンは膜の表面で重合して線維を形成しており、セプチン線維は管状に伸びた膜突起の周囲を取り巻くように並んでいた。膜突起部分の太さは約400nmと揃っており、これは丁度神経シナプス形成に重要なスパイン構造や精子鞭毛の付け根部分などと同じ大きさである。今回の発見により、今まで役割や機能が明確ではなかった第4の細胞骨格であるセプチンが実はダイナミックで細胞膜の形態制御に直接関与していることが明らかになった。
    本研究で用いたリポソームを利用した実験系によって、セプチンの活性を簡単に評価できることが示されたので、今まで困難だったセプチンの制御調節因子の探索も可能になった。

  20. Analysis of the mechanism of septin polymerization and depolymerization

    Grant number:18370077  2006 - 2007

    Grant-in-Aid for Scientific Research  Grant-in-Aid for Scientific Research(B)

    KINOSHITA Makoto

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    Authorship:Principal investigator  Grant type:Competitive

    Grant amount:\17620000 ( Direct Cost: \15400000 、 Indirect Cost:\2220000 )

  21. パーキンソン病およびアルツハイマー病で共通に凝集するセプチン蛋白質の解析

    Grant number:18659111  2006 - 2007

    文部科学省  科学研究費補助金(萌芽研究)  萌芽研究

    木下 専

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    Authorship:Principal investigator  Grant type:Competitive

    Grant amount:\3300000 ( Direct Cost: \3300000 )

    パーキンソン病におけるalpha-シヌクレインのリン酸化とレビー小体形成、アルツハイマー病におけるタウのリン酸化と神経原線維変化形成はいずれも家族性神経変性原因遺伝子産物を主成分とし、病態とよく相関することから、神経変性疾患の共通分子機構への手がかりを与えると考えられる。レビー小体形成に抑制的に作用するセプチンと、これら凝集体構成要素との相互作用に着目して、モデルマウスの作製・病態解析・バイオマーカー探索を行った。
    (1)alpha-シヌクレイン過剰発現マウスの病態が129位のセリン残基のリン酸化とよく相関することと、この両者がSept4の存在によって抑制される事実を見出した。今後はこの重要な分子間相互作用の詳細を明らかにし、治療への応用の可能性を探索する。
    (2)PD進行例ではドパミンニューロン内にリン酸化alpha-シヌクレインが増加する一方、Sept4が欠乏する。ヒト脳脊髄液中に微量検出されるSept4、リン酸化alpha-シヌクレイン、その他関連蛋白質の濃度の変動ないしこれらの組み合わせがPDによるドパミン神経障害の病勢や治療効果をモニターするバイオマーカーとなる可能性がある。そこで、これらの関連蛋白質のELISA系を開発できるよう、抗体の作製を行った。ELISA系の構築のため、神戸薬科大学との共同研究を開始した。
    (3)Sept4はPDのレビー小体の他、アルツハイマー病等で出現する神経原線維変化の副成分でもある。そこで、上記と同様にヒトtauP301Sを発現するtauopathyモデルマウス(吉山ら、 Neuron 2007)とSept4欠損マウスの交配を行った。。

  22. パーキンソン病におけるセプチンスカフォールドの破綻とドパミンニューロンの障害

    Grant number:18059017  2006 - 2007

    文部科学省  科学研究費補助金(特定領域研究)  特定領域研究

    木下 専

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    Authorship:Principal investigator  Grant type:Competitive

    Grant amount:\5200000 ( Direct Cost: \5200000 )

    (1)マウスの13種類のセプチン遺伝子のうち成熟脳特異的に発現するSept4を破壊し、網羅的行動解析によってドパミン神経伝達が減弱していることを見出した。これらのことから、ドパミン神経終末のセプチン系がドパミントランスポーターやtSNAREを安定化するスカフォールドとして機能することと、その破綻がパーキンソン病の病態に深く関わることが示唆された。
    (2)免疫電子顕微鏡(immunogold法)連続切片像から細胞形状と金粒子の分布情報を抽出してin silico3次元再構築を行い、シナプス近傍やグリア突起の特定の細胞膜ドメインの直下に集積する多様なセプチン・クラスターを同定した。これらの一部はシナプス近傍のグルタミン酸トランスポーターGLASTなどと共局在していた。一方、樹状突起棘起始部にあるものは構造的支持や隣接シナプス間の区画化に寄与する可能性がある。
    (3)培養神経細胞の細胞膜直下に集積したGFP-septinに対するFRAP(光退色後蛍光回復)法により、セプチン集合体の安定性が示された。このユニークな特性は細胞膜形状の保持や、膜蛋白質のスカフォールドないし拡散障壁としての役割に適したものと考えられる。RNAiでセプチンを枯渇させた細胞ではsyntaxin-1やGLASTのターンオーバーが亢進することから、(2)で同定されたセプチン・クラスターの一部は特定の膜蛋白質の局在や安定化に寄与するスカフォールドないし拡散障壁と推測される。また、(1)のSept4欠損マウスのドパミン神経ではこの機構が破綻するためにsyntaxin-1やドパミントランスポーターが欠乏するのかもしれない。

  23. 脳特異的Sept7欠損マウスを用いた神経系セプチンスカフォールドの機能解析

    Grant number:18022020  2006 - 2007

    日本学術振興会  科学研究費助成事業  特定領域研究

    木下 専

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    Authorship:Principal investigator  Grant type:Competitive

    Grant amount:\8400000 ( Direct Cost: \8400000 )

    マウス小脳Purkinje細胞樹状突起棘のくびれ部分(spine neck)、およびこれを取り巻くBergmann glial endfeetなど、細胞膜陥凹部直下に高度に集積するセプチン・クラスターを発見し、免疫電顕3次元再構築法で解析した(J Neurosci投稿中)。一方、大脳皮質初代培養ニューロンにおいて樹状突起棘のくびれ〜起始部に集積するセプチン・クラスターをRNAiで枯渇させると不安定なspineが多発することを示した(Curr Biol2007)。さらに、細胞膜直下のセプチン系が開口放出装置であるtSNAREsの安定化に寄与する拡散障壁であることをFRAP法(J Neurosci投稿中)およびSept4欠損マウスで示した(Neuron 2007)。しかし、Sept4欠損マウスの小脳異常は軽度であったため(Mol Cell Biol改訂中)、重複遺伝子の存在しないSept7遺伝子に着目してPurkinje細胞およびBergmann gliaにおける条件欠損(cK0)マウスの作製を行った。同研究班への公募参加が引き続き認められたので、H20-21年度中に行動・組織構築・微細形態・生化学的解析を完了する。

  24. Development of transgenic animal models of premature neurodegeneration

    2005.4 - 2006.3

    JST 

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  25. 「がん抑抑遺伝子」Sept4遺伝子破壊マウスの解析

    Grant number:17013048  2005

    文部科学省  科学研究費補助金(特定領域研究)  特定領域研究

    木下 専

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    Authorship:Principal investigator  Grant type:Competitive

    Grant amount:\4900000 ( Direct Cost: \4900000 )

    白血病や固形腫瘍におけるセプチン遺伝子異常が多数報告されている。しかし、その意義や分子機構はよくわかっていない(MCB2005)。培養細胞レベルの知見であるが、スタンフォード大学との共同研究により、細胞質分裂に先立つ染色体整列・分配にもセプチン系が必須であることを明らかにした。すなわち、セプチン系の破綻が染色体不安定化(異数性)につながり、がんの発生や悪性化に寄与する可能性がある(Science 2005)。セプチンサブユニットの1つSept4に関しては、ヒトの大腸がんにおいて高発現する一方、白血病においてはアポトーシス誘導活性によってがん抑制効果をもつという矛盾した仮説が異なるグループから提示されている。我々は、この問題を解決するために、Sept4遺伝子欠損マウスとSept4過剰発現マウスを作成して個体レベルの機能解析を行った。まず、いずれのマウス系統でも自然発がん傾向は見られないことを確定した。次に、Fasによる劇症肝炎モデルにおいて、Sept4欠損マウスでアポトーシス耐性が低いことが確定した。その原因を探索する過程で、Sept4蛋白質が肝実質細胞ではなく、肝炎や細胞外マトリックスの蓄積に重要な役割を果たす星細胞(Hepatic Stellate Cells, HSC)に特異的に発現することがわかった。このユニークな発現パターンを利用して、四塩化炭素による肝線維化モデルやジメチルニトロサミンによる発がんモデルにおけるHSC機能とSept4の存在意義を解析し、興味深い知見が得られた(祝迫ら、投稿準備中)。なお、本領域の主旨からは外れるが、生物学的に重要な知見も得られたので付記する。本マウスが予想外の雄性不妊を呈することが判明したため、精子鞭毛内の輪状小体がセプチンの環状高次集合体(セプチンリング)であり、その破綻によって生殖機能を失うこともわかった。同時にヒトの精子無力症検体でもセプチンリングの破綻を認め、セプチン細胞骨格の生殖医学的意義を発見した(Dev Cell 2005)。以上のような研究の進展によってセプチン系を再定義する機が熟し、総説を発表した(Curr Opin Cell Biol 2006)。

  26. 肝線維化と肝再生におけるアポトーシス関連遺伝子Septin4の機能解析

    Grant number:17659410  2005

    日本学術振興会  科学研究費助成事業  萌芽研究

    波多野 悦朗, 木下 専

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    Authorship:Coinvestigator(s)  Grant type:Competitive

    セプチンは重合性を持つGTP結合蛋白質で、多様な蛋白質や複合体を局在化ないしは安定化すると考えられる(Curr Opin Cell Biol 2006)。セプチン系の破綻によって細胞分裂に必須なチューブリンを主体とする染色体分配機構(紡錘体)とアクチンを主体とする細胞質分裂機構(収縮輪)が損なわれ、染色体不安定化や多核化をもたらすと考えられる(Genes Dev1997,Science 2005)。一方、「Sept4遺伝子に由来するアポトーシス誘導蛋白質ARTSはTGF-β依存的にミトコンドリアから核内に移行し、caspase-3を活性化しアポトーシスを誘導する」という、上記事実とは一見矛盾する仮説が提唱された(Larisch ら Nat Cell Biol 2001,EMBO J 2004)。我々は、「Sept4=アポトーシス誘導因子」仮説の検証を試みた。しかし、Fas抗体を用いた劇症肝炎モデルで、Sept4欠損マウスはヘテロタイプ、野生型と比較して、血清transaminaseの上昇およびHE染色における肝組織アポトーシス像に有意差を認めなかった。これらの結果から「Sept4=アポトーシス誘導因子」は否定的と考えられた。
    一方、マウス肝臓の免疫組織染色にてSept4は肝非実質細胞に発現を認めた。発現細胞を明らかにするためにマウス肝より密度勾配遠心法を用いて肝細胞、星細胞以外の間葉系細胞、肝星細胞を分離培養しそれぞれSept4の発現をWestern blottingにて検討した。その結果、Sept4は肝星細胞に特異的に発現していることが示唆された。星細胞の活性化は肝線維化をもたらすことより、肝線維化におけるSept4の役割を明らかにするために肝星細胞初代培養系およびマウス総胆管結紮による肝線維化モデルにて検討を進めている。

  27. 小脳バーグマングリア終末足に集積するセブチン系細胞骨格の機能解析

    Grant number:17024028  2005

    日本学術振興会  科学研究費助成事業  特定領域研究

    木下 専, 北野 歩

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    Authorship:Principal investigator  Grant type:Competitive

    Grant amount:\3600000 ( Direct Cost: \3600000 )

    セプチンサブユニットSept4の特異的な時空間的発現パターン(JCN 2000で報告済)に基づいてSept4遺伝子欠損マウスを作成し、組織学的解析と網羅的行動スクリーニングによって脳構築および脳機能の障害を探索した。その結果、幼若期の運動学習と成熟後の音刺激に対する驚愕反応のプレパルス抑制の2つのパラダイムで異常を見出した。そこで行動薬理学・組織学・電子顕微鏡学・細胞生物学・生化学的手法を用いた検討を重ねることにより、小脳発達過程におけるプルキンエ細胞や顆粒細胞に対するバーグマングリア細胞のニッシェ機能と黒質・線条体ドパミン系の高次生理機能におけるセプチン・スカフォールド系の重要性を発見・確立した(いずれも投稿中)。また、当該分野の主旨からは外れるが、本研究から医学・生物学的に重要な知見が派生したので付記する。本マウスが予想外の雄性不妊を呈することが判明したため、精子鞭毛内の輪状小体がセプチンの環状高次集合体(セプチンリング)であり、その破綻によって生殖機能を失うこともわかった。同時にヒトの精子無力症検体でもセプチンリングの破綻を認め、セプチン細胞骨格の臨床的意義を発見した(Dev Cell 2005)。
    以上のような研究の進展に基づいて、細胞生物学分野でセプチン系を再定義する総説を発表した(Curr Opin Cell Biol 2006)。

  28. Analysis of assembly and disassembly mechanisms of the septin cytoskeleton

    Grant number:16570158  2004 - 2005

    Grant-in-Aid for Scientific Research  Grant-in-Aid for Scientific Research(B)

    KINOSHITA Makoto

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    Grant amount:\3700000 ( Direct Cost: \3700000 )

    In a preceding international cooperative project, we reported biochemical interaction and possible functional interaction between mammalian septin complexes and the Cdc42 effector protein family, borgs (Nat.Cell Biol., 2001), however, the cellular functions of borgs have been unknown. To study whether the Cdc42-borg signaling system is involved in physiological regulation of the assembly and disassembly of septins, we raised antibody against a major borg species, revealed subcellular localization of the native borg protein, and depleted it by RNAi up to 90%. Unexpectedly, however, the depletion did not affect the localization and assembly of the septins and the major cytoskeleton (unpublished). To explore unidentified molecules that play major roles in the regulation of assembly and disassembly of the septin system, we purified septin-interacting molecules from cell extract and identified several septin-binders by a peptide mass fingerprinting method. We have revealed that a molecule involved in an important signal transduction pathway interact with septins both physically and functionally. We are currently testing the relevance of this interaction in tissue culture cells.
    In parallel with the above in vitro experiments, we attempted to reveal septin functions in vivo by creating mice lacking a septin gene. Through an unexpected phenotype of absolute male infertility of the septin-knockout mice, we found that a ring-shaped higher-order septin assembly in sperm flagellum (previously known as annulus) is essential for the structural and mechanical integrity of the spermatozoa. Using clinical materials collected under informed consent by urologists specialized in reproductive medicine, we found the ring-shaped higher-order septin assembly is lost in 25% of human asthenospemia, a male infertility syndrome with impaired flagellar motility (Dev.Cell 2005).
    Based on the above results, the researcher was invited as an author to review the progress of the septin biology (Curr.Opin.Cell Biol., 2006).

  29. セプチン系細胞骨格と脂質2重膜との相互作用の解析

    Grant number:16048214  2004

    文部科学省  科学研究費補助金(特定領域研究)  特定領域研究

    木下 専

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    Authorship:Principal investigator  Grant type:Competitive

    Grant amount:\3600000 ( Direct Cost: \3600000 )

    研究計画1:セプチン・リポソーム間の相互作用の解析
    組み換えセプチン複合体(野生型および各種変異体)とリポソームを作成・ゲルろ過精製し、電子顕微鏡で形状観察した。現在、両者の親和性を表面プラスモン共鳴法(Biacore)で測定するための条件検討を行っており、次年度以降報告予定である。
    研究計画2:細胞膜直下におけるセプチン集合体と細胞膜のダイナミックな相互作用の可視化と定量的解析
    培養細胞や遺伝子改変マウスの精子を用いて、細胞膜の裏打ちないし膜蛋白の拡散障壁の構成成分としてのセプチン系細胞骨格の機能を検討した。まず、哺乳類培養細胞の分裂時における染色体の赤道部への配列と分配、さらにそれに引き続く細胞質分裂において、微小管上から細胞膜直下のアクチン性収縮輪へとダイナミックにYFP-セプチンの局在が移行することをリアルタイム3D蛍光顕微鏡を用いて詳細に解析した。さらに、その局在のいずれもが細胞分裂に必須であることをRNAi技法を駆使して実証した。この研究により、細胞分裂において、セプチン細胞骨格が主要細胞骨格系と同様に重要かつ独自の役割を果たすことが明確に示された。次いで、精子尾部細胞膜において膜蛋白の拡散障壁として知られている輪状小体(annulus)の主成分がセプチンフィラメントの高次集合体であることを個体レベルで証明した(文献2)。この発見は精子形成の分子機構に迫るものであり、生物学的な興味に加えて臨床的にもヒトの男性不妊症研究に資する有意義な成果である。
    Sept4欠損マウス精子が輪状小体を持たないことを利用し、蛍光ラベルした脂質2重膜や膜蛋白の動態が正常コントロールとどのように異なるかをFRAP法を用いて解析するという課題が残った。

  30. 極低温電子線断層法によるセプチン系超分子構造体の解析

    2003.10 - 2007.3

    科学技術振興機構(JST)  戦略的創造研究推進事業PRESTO  個人型研究(さきがけ) 生体分子の形と機能

    木下 専

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  31. 細胞形態制御における細胞質骨格蛋白セプチンの機能解析

    Grant number:10213204  1998 - 2001

    日本学術振興会  科学研究費助成事業  特定領域研究(B)

    北山 仁志, 木下 専, 野田 亮

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    Grant amount:\44000000 ( Direct Cost: \44000000 )

    セプチンは真核生物の細胞分裂や細胞極性維持に必須なGTP結合タンパク質である。ヒトにおいては少なくとも9種類のセプチン・ファミリー・タンパク質が存在し、それらが複合体を形成してアクチン細胞骨格系や小胞分泌系と密接に関連することが判明しつつあるが(Longtine et al. Curr.Opin.Cell Biol.8,106-119,1996;Field and Kellogg, Trends Cell Biol.9,387-394,1999;Konoshita and Noda,2002)、その詳細な機能や作用機構は未知である。本研究課題では主に、セプチンと相互作用する分子の同定、アルツハイマー病患者および正常ラットの脳におけるセプチンの発現パターンの解析(木下ら、Am.J.Pathol,153,1551-1560,1998; J.Comp.Neurol,428,223-239.2000))、脳特異的な発現を示すH5のノックアウト・マウス作成を進めた。これまでに得られた成果として、セプチンがBorgファミリータンパク質と直接結合すること、Borgはセプチンの繊維構造を破壊し、Cdc42はこの活性を負に制御することが見出された(Jobertyら、Nature Cell Biol.2001)。すなわち、Cdc42がセプチンによる繊維状構造構築を正に制御する上流因子である可能性が示唆された。一方、ノックアウト・マウスの作成に関しては、これまでにマウス・ゲノムH5遺伝子の単離、ターゲテイィグ・ベクターの構築、相同組換えによるヘテロ・ノックアウトES細胞の樹立、良好なキメラマウスの作成までを進めた。今後、交配によるホモ欠損マウスの作成および表現型の解析を進める。

  32. Mechanisms of formation and destruction of the contractile ring during cytokinesis

    Grant number:10213101  1998 - 2001

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research  Grant-in-Aid for Scientific Research on Priority Areas

    MABUCHI Issei, HOSOYA Hiroshi, NUMATA Osamu, HAMAGUCHI Yukihisa, TANAKA Kazuma, KITAYAMA Hitoshi

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    We analyzed mechanism of formation of the contractile ring using various cells. In fission yeast cells, the contractile ring is formed from F-actin cables which accumulate at the medial region of the cell during nuclear division. The aster-like structure consisting of F-actin cables and the leading cable which elongates from the structure are important in this process. Cdcl2 plays a role in elongation of the leading cable, while Cdc15 plays a role in the fusion of the F-actin cables to the leading cable. We also analyzed function of Rho proteins in fission yeast, and found a novel G-protein Rho3. Rho3 is localized to the cell membrane .It was found that a novel formin family protein For3 is present downstream of either Rho3 or Cdc42. Rho3 and For3 controls both actin cytoskeleton and microtubules, and thereby regulate shape of the cell and position of cytokinesis. Cla4(PAK) and Bni1 (formin) cooperatively function in regulation of the septin ring in budding yeast. F-actin plays an important role in this process. In Tetrahymena, dimeric EF1a bundles F-actin. Ca/calmodulin dissociates this EF1a dimer and thus inhibit the bundling of F-actin. Tetrahymena fimbrin binds and bundles F-actin in a Ca-insensitive manner. It is localized to the cleavage furrow. We found that actin is less concentrated in the cortex near the centrosome while concentrated in the cortex away from it, where the furrow is formed, in the fourth division in sea urchin embryos. Role of microtubules in formation of the contractile ring in Xenopus eggs was analyzed. Disruption of microtubules by microtubule poisons did not interfere with contraction of the contractile ring, but interfered with its formation. We also found that the microtubules emanated from the both poles link with each other beneath the cleavage furrow. We tested a possibility that Ca ions play a role in cleavage signaling by analyzing Ca release around the cleavage furrow. Neither Ca waves nor small Ca releases such as Ca puffs or Ca blips were found at the leading edge of the cleavage furrow. Rho-kinase in HeLa cells binds to filamin A. Thus, it is possible that these proteins co-localize in the contractile ring.

  33. フラット・リバ-ジョン誘導遺伝子RECKによるがん転移抑制機構

    Grant number:10151224  1998

    文部科学省  科学研究費補助金(特定領域研究(A))  特定領域研究(A)

    高橋智聡, 糸原重美, 高橋雅英, 野田亮

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    RECK遺伝子はras癌遺伝子でトランスフォ-ムした細胞の形質を正常形質に復帰させる活性を指標として発現クロ-ニング法により単離された遺伝子である。RECK遺伝子はほとんどすべての正常組織で発現しているが、癌細胞や種々の癌遺伝子でトランスフォ-ムした細胞では発現が著しく抑制されている。この遺伝子を転移性の高い細胞(B16やHT1080)で強制発現させると、浸潤・転移能の著明な抑制が起こる。本研究は、野生型及び変異を導入したRECK蛋白質の機能解析、RECK遺伝子プロモ-タ-領域の解析、RECK遺伝子欠損マウスの作製と表現型解析、及び、種々のステ-ジの癌組織の免疫組織学的観察等を行い、RECK遺伝子の生理学的機能及び作用機構、さらには、発がんとがんの悪性化に於ける役割について洞察を加えることを目的として開始され、以下の諸点を明らかにした。
    1. RECK遺伝子がコ-ドする蛋白質は分子量約110kDaの膜アンカ-糖蛋白質であり、Matrix Metalloprotease(MMP)-9と直接に結合してその分泌を負に制御する。また、MMP-2の中間活性化型から活性化型への転換を阻害する。
    2. セリン・プロテア-ゼ・インヒビタ-様領域中に存在するアスパラギン酸の置換は、RECK蛋白質のMMP制御活性を低下させる。
    3. マウスRECK遺伝子のプロモ-タ-領域にはSp1/Sp3転写因...

  34. セプチンフィラメントによるアルツハイマー病動物モデルの作成に関する研究

    Grant number:199700699A  1997.4 - 1998.3

    厚生省(厚生労働省)  厚生科学研究費補助金 脳科学研究推進特別事業(若手研究者特別枠) 

    木下 専

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    Authorship:Principal investigator  Grant type:Competitive

    Grant amount:\5000000

  35. 電気刺激後の脳に発現誘導される分泌性および膜結合性神経再生関連蛋白の探索

    Grant number:09780722  1997 - 1998

    日本学術振興会  科学研究費助成事業  奨励研究(A)

    木下 専

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    Authorship:Principal investigator  Grant type:Competitive

    Grant amount:\2000000 ( Direct Cost: \2000000 )

    神経再生や可塑性に関与する分泌性ないし膜結合性蛋白の探索を目的として、電気刺激後のマウス脳由来のcDNAライブラリーをシグナルシークエンス・トラップ法によってスクリーニングした。これにより単離された既知遺伝子はBMP2 receptor,somatostatin receptor4,glycine receptor α1,L1,neuronatin-3,IGF-bindingprotein5等多数であった。一方、ESTのみに類似性を有する遺伝子は12クローン、データベース上の遺伝子と全く類似性のない遺伝子は16クローンであった。全長が単離できた未知遺伝子のうち、培養細胞に発現させると細胞接着抑制活性を有する分子量27.8kDのI型膜蛋白(eludin-1と仮称)をコードするものに集中して解析を進めた。
    1) ヒトのESTデータベースにcludin-1と類似した27.5kDのI型膜蛋白をコードするeludin-2を見出し、同様の抗細胞接着活性を有することを確認した。
    2) eludin-1-GFP融合蛋白も同等の活性を有し、細胞膜に集積することを確認した。
    3) eludin-1を高発現した線維芽細胞細胞においては接着斑の数が極端に減少していることを見出した。
    4) eludin-1の接着斑形成阻害・抗接着活性における、各種プロテインキナーゼ阻害剤の効果をスクリーニングした。このうちstaurosporinによって活性がブロックされることから、リン酸化・脱リン酸化による活性制御の可能性を見出した。
    5) eludin-1の抗細胞接着活性は、collagenないしfibronectinを基質にコーティングすることで拮抗されたことから、細胞外マトリックスとの相互作用が重要であることを確認した。

  36. 収縮輪に局在する新規細胞骨格蛋白Nedd5の解析

    Grant number:08259205  1996

    文部科学省  科学研究費補助金(重点領域研究)  重点領域研究

    木下 専

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    Authorship:Principal investigator  Grant type:Competitive

    Grant amount:\2000000 ( Direct Cost: \2000000 )

    出芽酵母およびショウジョウバエのseptinは収縮輪に局在するフィラメントの構成成分であり、その機能喪失型変異体は不完全な細胞質分裂により多核化することが知られている。
    研究代表者らは、本研究課題において、
    1)マウス胚神経系で高発現する遺伝子としてcDNAサブトラクション法により単離したNedd5が、septinファミリーに属し、特徴的なGTPaseドメインを有する41.5kDの蛋白をコードすることを示した。
    2)Nedd5がヒトおよびマウス細胞の収縮輪に局在し、抗Nedd5抗体の微小注入によりヒト癌細胞HeLaの細胞質分裂を阻害することを示した。
    3)Nedd5‐green fluorescent protein融合蛋白がanaphaseにおいて赤道部に帯状に局在し、細胞質分裂期に収縮輪近傍に濃縮し、さらにはmidbodyに局在することを、HeLa細胞における連続観察で確認した。
    GTPaseドメインに変異を導入し、GTP結合活性を欠くNedd5は、HeLa細胞において正常に局在しないのみならず、内在性Nedd5の分布も阻害することを示した。
    5)細胞質分裂において重要なアクチンやRhoと相互作用する可能性を探るため、これらの特異的阻害剤cytochalasin DまたはC3 toxinをHeLa細胞に作用させると、Nedd5が不規則に凝集するとともに、細胞質分裂が停止することを示した。
    以上より研究代表者らは、哺乳動物細胞の細胞質分裂におけるseptin、Nedd5の重要性を明らかにするとともに、分裂間期におけるNedd5に関しても興味深い知見を得た(投稿中Kinoshita et al. : A mammalian septin,Nedd5,is a novel cytoskeletal component interacting with rhoactin system.)。

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Teaching Experience (On-campus) 126

  1. Bioscience Laboratory

    2020

  2. 生体構築論

    2020

  3. 生物学実験

    2020

  4. Biology Experiment

    2020

  5. 生物学基礎I

    2020

  6. Bioscience Laboratory

    2019

  7. 生体構築論

    2019

  8. 卒業実験

    2019

  9. 生物科学実験IV

    2019

  10. 情報機構学特論2

    2019

  11. 情報機構学講究2

    2019

  12. 生物学実験

    2019

  13. Cell Biology 3

    2019

  14. Biology Experiment

    2019

  15. 生物学基礎I

    2019

  16. 遺伝学Ib

    2019

  17. 分子生物学演習Ⅰ

    2019

  18. Bioscience Laboratory

    2018

  19. Bioscience Laboratory

    2018

  20. 分子生物学演習Ⅰ

    2018

  21. 生体構築論

    2018

  22. 卒業実験

    2018

  23. 情報機構学特論2

    2018

  24. 情報機構学講究2

    2018

  25. 生物学実験

    2018

  26. Cell Biology 3

    2018

  27. 学部向け集中講義

    2018

     詳細を見る

    非常勤講師:北海道大学 電子科学研究所 生命科学研究部門 光細胞生理研究分野 根本 知己 教授

  28. Biology Experiment

    2018

  29. 生物学基礎I

    2018

  30. Special Lecture Tokuron, Neuroscience Course, Nagoya Univ Graduate School of Medicine

    2018

     詳細を見る

    Exploring pre-/post-/peri-synaptic functions of the septin cytoskeleton

  31. Bioscience Laboratory

    2017

  32. 分子生物学演習Ⅰ

    2017

  33. 生体構築論

    2017

  34. 卒業実験

    2017

  35. 情報機構学特論2

    2017

  36. 情報機構学講究2

    2017

  37. 生物学実験

    2017

  38. Cell Biology 3

    2017

  39. Biology Experiment

    2017

  40. 生物学基礎I

    2017

  41. Bioscience Laboratory

    2016

  42. 分子生物学演習Ⅰ

    2016

  43. 生体構築論

    2016

  44. 卒業実験

    2016

  45. 情報機構学特論2

    2016

  46. Biology Experiment

    2016

  47. 生物学実験

    2016

  48. Cell Biology 3

    2016

  49. 生物学基礎I

    2016

  50. Bioscience Laboratory

    2015

  51. 分子生物学演習Ⅰ

    2015

  52. 生体構築論

    2015

  53. 卒業実験

    2015

  54. 情報機構学特論2

    2015

  55. Biology Experiment

    2015

  56. 生物学実験

    2015

  57. Cell Biology 3

    2015

  58. 学部向け集中講義

    2015

     詳細を見る

    非常勤講師:京都大学 再生医科学研究所 附属ナノ再生医工学研究センター バイオメカニクス研究領域 安達 泰治 教授

  59. 生物学入門

    2015

  60. 生物学基礎I

    2015

  61. Bioscience Laboratory

    2014

  62. 分子生物学演習Ⅰ

    2014

  63. 生体構築論

    2014

  64. 卒業実験

    2014

  65. 情報機構学特論2

    2014

  66. 情報機構学講究2

    2014

  67. 生物学実験

    2014

  68. Cell Biology 3

    2014

  69. 学部向け集中講義

    2014

     詳細を見る

    非常勤講師: 国立がん研究センター研究所 分子細胞治療研究分野 落谷 孝広 分野長

  70. Biology Experiment

    2014

  71. 生物学入門

    2014

  72. Bioscience Laboratory

    2013

  73. 情報機構学特論2

    2013

  74. 分子生物学演習Ⅰ

    2013

  75. 生体構築論

    2013

  76. 卒業実験

    2013

  77. 情報機構学講究2

    2013

  78. 生物学実験

    2013

  79. Cell Biology 3

    2013

  80. 学部向け集中講義

    2013

     詳細を見る

    非常勤講師: 京都大学 iPS細胞研究所(CIRA) 臨床応用研究部門 井上 治久 准教授

  81. Biology Experiment

    2013

  82. 生物学入門

    2013

  83. 分子生物学演習Ⅰ

    2012

  84. 生体構築論

    2012

  85. 生物科学実験Ⅴ

    2012

  86. 基礎遺伝学II

    2012

  87. 生命化学I

    2012

  88. 生物学実習

    2012

  89. 生物学入門

    2012

  90. Bioscience Laboratory

    2012

  91. 情報機構学特論2

    2012

  92. 卒業実験

    2012

  93. 情報機構学講究2

    2012

  94. 生物学入門

    2012

  95. 生物学実験

    2012

  96. Cell Biology 3

    2012

  97. 学部向け集中講義

    2012

     詳細を見る

    非常勤講師:理化学研究所 発生・再生科学総合研究センター 笹井 芳樹 グループディレクター

  98. Biology Experiment

    2012

  99. 医学系研究科グローバルCOEプログラム「機能分子医学への神経疾患・腫瘍の融合拠点」

    2012

     詳細を見る

    遺伝子改変マウスを用いたセプチン細胞骨格系の神経機能と病態へのアプローチ

  100. Introduction to Biology

    2011

  101. First Year Seminar A

    2011

  102. 生物学実習

    2011

  103. 分子生物学演習Ⅰ

    2011

  104. 生体構築論

    2011

  105. 生物科学実験Ⅴ

    2011

  106. 生命化学I

    2011

  107. 情報機構学特論2

    2011

  108. 卒業実験

    2011

  109. 情報機構学講究2

    2011

  110. 学部向け集中講義

    2011

     詳細を見る

    非常勤講師:大阪大学大学院 生命機能研究科 木村 宏 准教授

  111. 生物学実験

    2011

  112. 学部向け集中講義

    2011

     詳細を見る

    非常勤講師:藤田保健衛生大学 総合医科学研究所 宮川 剛 教授

  113. 分子生物学演習Ⅰ

    2010

  114. 生命化学Ⅰ

    2010

  115. 生物学入門

    2010

  116. 生物科学実験Ⅴ

    2010

  117. 生物学実習

    2010

  118. 生体構築論

    2010

  119. 情報機構学特論2

    2010

  120. 卒業実験

    2010

  121. 情報機構学講究2

    2010

  122. 学部向け集中講義

    2010

     詳細を見る

    非常勤講師:藤田保健衛生大学 総合医科学研究所 宮川 剛 教授

  123. 生体構築論

    2009

  124. 分子遺伝学I

    2009

  125. 卒業実験

    2009

  126. 生命化学Ⅰ

    2009

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Social Contribution 3

  1. 大学での生命科学の現状と高校・中学「生物」に求められる新課程への対応

    Role(s):Lecturer, Advisor, Informant

    四天王寺学園 中学校・高等学校  2016.12

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    Audience: Teachers

    Type:Seminar, workshop

  2. 藤田保健衛生大学共同利用・共同研究拠点運営委員会

    2015

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    拠点運営への助言および評価

  3. 脳と心はどこまで理解できるようになったか?

    Role(s):Appearance, Lecturer, Informant

    愛知県立時習館高等学校  オープンフォーラム  2013.10

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    Audience: Teachers, General

    Type:Lecture

Media Coverage 10

  1. ユビキチン修飾系、オートファジーに次ぐ新しいUBL3翻訳後修飾系を世界で初めて発見 Internet

    日本の研究.com  日本の研究.comニュース  【注目プレスリリース】  2018.9

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    Author:Other 

  2. 神経伝達物質グルタミン酸をシナプスから浄化するしくみに迫る Internet

    名古屋大学ウェブサイト  MONOist 医療技術ニュース  2015.12

  3. Sirt1 overexpression in neural and vascular endothelial cells Promotional material

    RIKEN BioResource Center  BRC News  Mouse of the Month  2015.2

  4. 長寿遺伝子サーチュインを働かせて認知症予防~動脈硬化による認知症の新たな治療法確立に寄与すると期待~ Internet

    国立循環器病研究センターウェブサイト  2014.9

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    Author:Other 

  5. 長寿遺伝子産物SIRT1(サーチュイン)の活性化で神経難病ALSマウスが延命―神経難病の治療法開発へ期待― Internet

    名古屋大学ウェブサイト  2014.9

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    Author:Myself 

  6. 神経線維の伸長 セプチンが不可欠 Newspaper, magazine

    日経産業新聞  日経産業新聞  2013.10

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    Author:Other 

  7. 神経細胞つくる細胞骨格 Newspaper, magazine

    中日新聞  中日新聞  2013.10

  8. 神経線維の伸長を促進する微小管の「脱」安定化機構の発見 Internet

    名古屋大学ウェブサイト  2013.10

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    Author:Myself 

  9. 細胞形状の変形過程解明 たんぱく質セプチン関与顕微鏡で可視化 Newspaper, magazine

    日刊工業新聞  日刊工業新聞  2009.1

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    Author:Other 

  10. 神経細胞つくるタンパク質 セプチンの機能解析 Newspaper, magazine

    中日新聞  中日新聞  2009.1

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    Author:Other 

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