Develpment of imaging technology for structural dynamics of synapses

• Project Area: Resonance Biology for Innovative Bioimaging
• Project/Area Number: 18H04727
• Research Category: Grant-in-Aid for Scientific Research on Innovative Areas (Research in a proposed research area)
• Allocation Type: Single-year Grants
• Research Institution: The University of Tokyo
• Principal Investigator: 岡部 繁男 東京大学, 大学院医学系研究科(医学部), 教授 (60204012)
• Project Period (FY): 2018-04-01 – 2020-03-31
• Project Status: Completed (Fiscal Year 2019)
• Budget Amount: ¥10,010,000 (Direct Cost: ¥7,700,000、Indirect Cost: ¥2,310,000); Fiscal Year 2019: ¥5,070,000 (Direct Cost: ¥3,900,000、Indirect Cost: ¥1,170,000); Fiscal Year 2018: ¥4,940,000 (Direct Cost: ¥3,800,000、Indirect Cost: ¥1,140,000)
• Keywords: 分子神経生物学 / イメージング / 超解像技術 / シナプス / 分子動態

Outline of Annual Research Achievements

本研究では神経ネットワークの要であるシナプスの形成や維持の分子機構を探索するために必須の技術である、樹状突起スパインの形態と分子動態を定量的に測定するための技術開発を実施した。具体的な研究として（１）厚い標本での樹状突起スパイン形態の高解像度画像取得を目指した技術開発、（２）多数のスパイン形態データを定量的・客観的に解析するための手法の開発、（３）分子動態に関するデータとスパイン形態のデータを統合するための方法論の探索の３項目を行った。（１）については、これまで用いてきた構造化照明の手法に加えて、走査型レーザー顕微鏡で超解像を得る手法と組織の透明化と拡大手法を組み合わせることで、スパインの微細構造のデータを組織標本から得ることに成功した。（２）についてはスパイン形態の特徴を多次元空間内で表現し、かつスパイン形態の時間的な変化をこれに加えて解析することにより、新しいスパイン分類を提案した。この新しい分類はスパインの成長、安定化、消失過程を考慮したものであり、従来の分類よりも生物学的な意味が明確であるという利点がある。（３）については蛍光相関のシグナルをスパイン内部の微小体積から得ることで、細胞質内でアクチン細胞骨格が拡散を阻害する主要な要素であること、この拡散障壁が可塑性誘導刺激の際に数分間だけ緩和することで、高分子量のシグナル分子のスパイン内部での動きが高まることが明らかになった。これらの微細形態と分子動態を結び付ける方法論の開発によりスパインの形態と内部での分子動態を統合した理解が今後更に進展することが期待される。

Research Progress Status

令和元年度が最終年度であるため、記入しない。

Strategy for Future Research Activity

令和元年度が最終年度であるため、記入しない。

Research Products

• [Int'l Joint Research] Edinburgh University(英国)
• [Int'l Joint Research] University of Massachusetts(米国)
• [Journal Article] Vasodilator-stimulated phosphoprotein (VASP) is recruited into dendritic spines via G-actin-dependent mechanism and contributes to spine enlargement and stabilization. (2020)
  • Author(s): Iwasaki K, Obashi K, Okabe S.
  • Journal Title: European Journal of Neuroscience Volume: 51 Issue: 3 Pages: 806-821
  • DOI: 10.1111/ejn.14634
  • Peer Reviewed
• [Journal Article] Impaired actin dynamics and suppression of Shank2-mediated spine enlargement in cortactin knockout mice (2020)
  • Author(s): Tanaka Shinji、Masuda Yasutaka、Harada Akihiro、Okabe Shigeo
  • Journal Title: Microscopy Volume: 69 Issue: 1 Pages: 44-52
  • DOI: 10.1093/jmicro/dfaa001
  • Peer Reviewed / Open Access
• [Journal Article] Precise temporal regulation of molecular diffusion within dendritic spines by actin polymers during structural plasticity. (2019)
  • Author(s): Obashi, K., Matsuda, A., Inoue, Y., Okabe, S.
  • Journal Title: Cell Reports Volume: 27 Issue: 5 Pages: 1503-1515
  • DOI: 10.1016/j.celrep.2019.04.006
  • Peer Reviewed / Open Access
• [Journal Article] Tumor suppressor protein CYLD regulates morphogenesis of dendrites and spines. (2019)
  • Author(s): Li, J., Sekine-Aizawa, Y., Ebrahimi, S., Tanaka, S., Okabe, S.
  • Journal Title: European Journal of Neuroscience Volume: 50 Issue: 4 Pages: 2722-2739
  • DOI: 10.1111/ejn.14421
  • Peer Reviewed
• [Journal Article] Neonatal social isolation increases the proportion of the immature spines in the layer 2/3 pyramidal neurons of the somatosensory cortex. (2019)
  • Author(s): Iguchi, R., Tanaka, S., Okabe, S.
  • Journal Title: Neuroscience Research Volume: 154 Pages: 27-34
  • DOI: 10.1016/j.neures.2019.05.004
  • Peer Reviewed
• [Journal Article] Temporal sequences of synapse disintegration triggered by afferent axon transection - time-lapse imaging study of pre- and post-synaptic molecules. (2019)
  • Author(s): Cho T, Kashiwagi Y, Okabe S.
  • Journal Title: eNeuro Volume: 6 Issue: 5 Pages: ENEURO.0459-18.2019
  • DOI: 10.1523/eneuro.0459-18.2019
  • Peer Reviewed / Open Access
• [Journal Article] Nano-scale analysis of synapse morphology in an autism mouse model with 15q11-13 copy number variation using focused ion beam milling and scanning electron microscopy (2019)
  • Author(s): Sato, Y., and S.Okabe
  • Journal Title: Microscopy Volume: 68 Issue: 2 Pages: 122-132
  • DOI: 10.1093/jmicro/dfy128
  • Peer Reviewed
• [Journal Article] Cellular cartography of the organ of Corti based on optical tissue clearing and machine learning (2019)
  • Author(s): Urata Shinji、Iida Tadatsune、Yamamoto Masamichi、Mizushima Yu、Fujimoto Chisato、Matsumoto Yu、Yamasoba Tatsuya、Okabe Shigeo
  • Journal Title: eLife Volume: 8
  • DOI: 10.7554/elife.40946
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Ultrastructural Observation of Glutamatergic Synapses by Focused Ion Beam Scanning Electron Microscopy (FIB/SEM) (2019)
  • Author(s): Takahashi-Nakazato Ai、Parajuli Laxmi Kumar、Iwasaki Hirohide、Tanaka Shinji、Okabe Shigeo
  • Journal Title: Methods in Molecular Biology Volume: 1941 Pages: 17-27
  • DOI: 10.1007/978-1-4939-9077-1_2
  • ISBN: 9781493990764, 9781493990771
  • Peer Reviewed
• [Journal Article] LAMP5 in presynaptic inhibitory terminals in the hindbrain and spinal cord: a role in startle response and auditory processing (2019)
  • Author(s): .Koebis, M., Urata, S., Shinoda, Y., Okabe, S., Yamasoba, T., Nakao, K., Aiba, A., and T. Furuichi,
  • Journal Title: Molecular Brain Volume: 12 Issue: 1 Pages: 20-20
  • DOI: 10.1186/s13041-019-0437-4
  • Peer Reviewed / Open Access
• [Journal Article] Computational geometry analysis of dendritic spines by structured illumination microscopy (2019)
  • Author(s): Kashiwagi, Y., Higashi, T., Obashi, K., Sato, Y., Komiyama, N., Grant, S. G. N. and S. Okabe
  • Journal Title: Nature Communications Volume: 10 Issue: 1 Pages: 1285-1285
  • DOI: 10.1038/s41467-019-09337-0
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Microglia permit climbing fiber elimination by promoting GABAergic inhibition in the developing cerebellum (2018)
  • Author(s): Nakayama Hisako、Abe Manabu、Morimoto Chie、Iida Tadatsune、Okabe Shigeo、Sakimura Kenji、Hashimoto Kouichi
  • Journal Title: Nature Communications Volume: 9 Issue: 1 Pages: 2830-2830
  • DOI: 10.1038/s41467-018-05100-z
  • Peer Reviewed / Open Access
• [Journal Article] The inflammasome-derived cytokine IL18 suppresses amyloid-induced seizures in Alzheimer’s prone mice (2018)
  • Author(s): Tzeng, T-C, Hasegawa, Y., Iguchi, R., Caffrey, D. R., Thatcher, E. J., Mao, W.J., Germain, G., Tamburro, N. D., Okabe, S., Heneka, M., Latz, E., Futai, K.,and D. T. Golenbock
  • Journal Title: Proc. Natl. Acad. Sci. USA Volume: 115 Issue: 36 Pages: 9002-9007
  • DOI: 10.1073/pnas.1801802115
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Red-Shifted Fluorogenic Substrate for Detection of lacZ-Positive Cells in Living Tissue with Single-Cell Resolution (2018)
  • Author(s): Ito Hiroki、Kawamata Yu、Kamiya Mako、Tsuda-Sakurai Kayoko、Tanaka Shinji、Ueno Tasuku、Komatsu Toru、Hanaoka Kenjiro、Okabe Shigeo、Miura Masayuki、Urano Yasuteru
  • Journal Title: Angewandte Chemie International Edition Volume: 57 Issue: 48 Pages: 15702-15706
  • DOI: 10.1002/anie.201808670
  • Peer Reviewed
• [Journal Article] Spatial impact of microglial distribution on dynamics of dendritic spines (2018)
  • Author(s): Iida, T., Tanaka, S., and S. Okabe
  • Journal Title: European Journal of Neuroscience Volume: NA Issue: 11 Pages: 1400-1417
  • DOI: 10.1111/ejn.14325
  • Peer Reviewed
• [Presentation] Morphological analysis of dendritic spines and microglia in cortactin knockout mice (2020)
  • Author(s): Y. Masuda, S. Tanaka, S. Okabe
  • Organizer: 第125 回日本解剖学会総会・学術集会
• [Presentation] Super-resolution microscopy-based computational analysis of synaptic structure (2020)
  • Author(s): Y. Kashiwagi, S. Okabe
  • Organizer: 第125 回日本解剖学会総会・学術集会
• [Presentation] AiCE mouse: a novel tool to probe LTP-related change in dendritic spines (2019)
  • Author(s): K. Kuboyama, T. Inoue, Y. Hashimotodani, T. Itoh, T. Suzuki, Y. Ohtsuka, R. Kinoshita, P. Gusain, M. Kano, S. Okabe, M.K. Yamada
  • Organizer: 第42回日本神経科学大会
• [Presentation] Comprehensive gene expression profiling between BTBR mice, a mouse model of autism spectrum disorder, and C57BL6/J showing high levels of sociality (2019)
  • Author(s): S. Mizuno, J. Hirota, H. Iwasaki, S. Okabe, Y. Sano, T. Furuichi
  • Organizer: 第42回日本神経科学大会
• [Presentation] Computational geometry analysis of dendritic spines by structured illumination microscopy (2019)
  • Author(s): Y. Kashiwagi, T. Higashi, K. Obashi, S. Okabe
  • Organizer: Japan-US Neuroscience Workshop Current trends and future directions of synapse-circuit plasticity research
  • Int'l Joint Research / Invited
• [Presentation] Nanostructure and dynamics of dendritic spines (2019)
  • Author(s): S. Okabe
  • Organizer: 2019 Annual Meeting of Korean Association of Anatomists
  • Int'l Joint Research / Invited
• [Presentation] Nanostructure and molecular dynamics of dendritic spines (2019)
  • Author(s): S. Okabe
  • Organizer: Resonance Bio International Symposium
  • Int'l Joint Research
• [Presentation] Nanoscale analysis of neuronal spine synapses (2019)
  • Author(s): S. Okabe
  • Organizer: The 20th International Conference on Systems Biology (ICSB2019)
  • Int'l Joint Research / Invited
• [Presentation] 海馬分散培養細胞における脳由来神経栄養因子(BDNF)の放出の解析 (2019)
  • Author(s): 福田裕太、柏木有太郎、岡部繁男
  • Organizer: 第124回日本解剖学会総会・学術集会
• [Presentation] Neonatal social isolation induces immature spines in layer 2/3 neurons of the somatosensory cortex in vivo (2019)
  • Author(s): 井口理沙、田中慎二、岡部繁男
  • Organizer: 第124回日本解剖学会総会・学術集会
• [Presentation] イメージングによる脳神経回路の発達とその障害の理解 (2018)
  • Author(s): 岡部繁男
  • Organizer: 第７７回日本医学放射線学会総会
  • Invited
• [Presentation] Synapse elimination triggered by BMP4 exocytosis and presynaptic BMP4 receptor activation (2018)
  • Author(s): Takahito Higashi, Shinji Tanaka, Tadatsune Iida, Shigeo Okabe
  • Organizer: 第70回日本細胞生物学会 第51回日本発生生物学会 合同大会
• [Presentation] Rapid clearing and labeling of mouse cochlea by modified scaleS enable exhaustive analysis of hair cell (2018)
  • Author(s): Shinji Urata, Tadatsune Iida, Yu Mizushima, Chisato Fujimoto, Yu Matsumoto, Tatsuya Yamasoba, Shigeo Okabe
  • Organizer: 第70回日本細胞生物学会 第51回日本発生生物学会 合同大会
• [Presentation] ACF7, an actin-microtubule crosslinking protein, stably associates with postsynaptic sites (2018)
  • Author(s): Yutaro Kashiwagi, Shigeo Okabe
  • Organizer: 第70回日本細胞生物学会 第51回日本発生生物学会 合同大会
• [Presentation] Spatial relationship between microglia and synapse stability studied by in vivo imaging (2018)
  • Author(s): Shinji Tanaka, Tadatsune Iida, Shigeo Okabe
  • Organizer: 第70回日本細胞生物学会 第51回日本発生生物学会 合同大会
• [Presentation] Synapse remodeling in vitro and in vivo (2018)
  • Author(s): Shigeo Okabe
  • Organizer: Sattelie symposium for the 41th Annual Meeting of the Japan Neuroscience Society “Frontiers of Neurointelligence”
  • Int'l Joint Research / Invited
• [Presentation] Neurotecnologly innovation from the bottom up: strategies for product development at major brain research initiatives (2018)
  • Author(s): Shigeo Okabe
  • Organizer: Minding Neurotechnology: Delivering responsible innovation for health and well being
  • Int'l Joint Research / Invited
• [Presentation] Japan Brain/MINDS (2018)
  • Author(s): Shigeo Okabe
  • Organizer: Global Neuroethics Summit
  • Int'l Joint Research / Invited
• [Presentation] Imaging synapse remodeling in vitro and in vivo (2018)
  • Author(s): Shigeo Okabe
  • Organizer: Joint Conference of 8th Asia Pacific International Congress of Anatomists and 68th Korean Association of Anatomists
  • Int'l Joint Research / Invited
• [Remarks] 東京大学大学院医学系研究科神経細胞生物学分野
  • URL: http://synapse.m.u-tokyo.ac.jp/
• [Remarks] http://synapse.m.u-tokyo.ac.jp/index.html