Advanced technology development for analyzing molecular dynamics and structure in synapse formation

• Project/Area Number: 26640039
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Neurochemistry/Neuropharmacology
• Research Institution: Shinshu University
• Principal Investigator: UEMURA Takeshi 信州大学, 学術研究院医学系, 准教授 (00372368)
• Co-Investigator(Renkei-kenkyūsha): SATO Chikara 独立行政法人産業技術総合研究所, 脳神経情報研究部門, 研究グループ長 (00357146)
• Project Period (FY): 2014-04-01 – 2016-03-31
• Project Status: Completed (Fiscal Year 2015)
• Budget Amount: ¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000); Fiscal Year 2015: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000); Fiscal Year 2014: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000)
• Keywords: 脳・神経 / 神経科学 / シナプス

Outline of Final Research Achievements

Elucidation of synapse formation process is essential for understanding synaptic dysfunction in neurodevelopmental disorders. Atmospheric scanning electron microscope (ASEM) is an electron microscope which was developed recently to directly observe subcellular structures and organelles through a silicon nitride (SiN) film under atmospheric pressure. In this study, we cultured neurons on poly-L-lysine and laminin coated ASEM dish with the SiN film. Presynaptic differentiation was induced by extracellular domain of synapse organizer-coated magnetic beads and analyzed by fluoresence microscope and ASEM. Fluorescence signals for presynaptic proteins were accumulated on the beads particularly concentrated at the site of contact with axon. Subsequent ASEM analysis revealed that presynaptic protein, presumably of an indistinguishable neurite branch, surrounded each beads. Thus, ASEM is a potentially useful tool for analysis of molecular localization and fine structure of synapse.

Research Products

• [Journal Article] Mechanisms of splicing-dependent trans-synaptic adhesion by PTPδ-IL1RAPL1/IL-1RAcP for synaptic differentiation (2015)
  • Author(s): Yamagata A., Yoshida, T., Sato, Y., Goto-Ito, S., Uemura, T., Maeda, A., Shiroshima, T., Iwasawa-Okamoto, S., Mori, H., Mishina, M., Fukai , S.
  • Journal Title: Nature Communications Volume: 6 Issue: 1 Pages: 6926-6926
  • DOI: 10.1038/ncomms7926
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Electron microscopy of primary cell cultures in solution and correlative optical microscopy using ASEM (2014)
  • Author(s): K.Hirano, T.Kinoshita, T.Uemura, H.Motohashi, Y.Watanabe, T.Ebihara, H.Nishiyama, M.Sato, M.Suga, Y.Maruyama, N.M.Tsuji, M.amamoto, S.Nishihara, C.Sato
  • Journal Title: Ultramicroscopy Volume: 143 Pages: 52-66
  • DOI: 10.1016/j.ultramic.2013.10.010
  • Peer Reviewed / Acknowledgement Compliant
• [Presentation] Insights into structure and assembly of GluD2-Cbln1-neurexin adhesion complex for cerebellar synapse formation (2015)
  • Author(s): 植村健
  • Organizer: 第８８回日本生化学大会
  • Place of Presentation: 神戸ポートアイランド（兵庫県神戸市）
  • Year and Date: 2015-12-01
  • Invited
• [Presentation] 小脳シナプス形成を担うGluRδ2-Cbln1-neurexin接着分子複合体の構造基 (2015)
  • Author(s): 植村健
  • Organizer: 第53回日本生物物理学会年会
  • Place of Presentation: 金沢大学（石川県金沢市）
  • Year and Date: 2015-09-14
  • Invited
• [Presentation] The Atmospheric Scanning Electron Microscope (ASEM) Observes Axonal Segmentation and Synaptic Induction in Solution (2014)
  • Author(s): 佐藤 主税, 木下 貴明, 植村 健, 平野 和己, 川田 正晃, 西山英利, 佐藤 真理, 海老原 達彦, 須賀三雄, 西原祥子
  • Organizer: 2014 Microscopy & Microanalysis meeting
  • Place of Presentation: ハートフォード, アメリカ合衆国
  • Year and Date: 2014-08-05
• [Presentation] 受容体でコートした磁気ビーズによる初代培養シナプス形成誘導のASEM水中免疫電顕 (2014)
  • Author(s): 植村 健, 佐藤 真理, 西山 英利, 須賀 三雄, 佐藤 主税
  • Organizer: 日本顕微鏡学会 第70回記念学術講演
  • Place of Presentation: 幕張メッセ国際会議場, 千葉
  • Year and Date: 2014-05-11