High spatiotemporal resolution analyses of dynamics of synaptic vesicle proteins

• Project/Area Number: 18H02526
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Review Section: Basic Section 46010:Neuroscience-general-related
• Research Institution: Kyoto University
• Principal Investigator: Hirano Tomoo 京都大学, 理学研究科, 教授 (50181178)
• Project Period (FY): 2018-04-01 – 2021-03-31
• Project Status: Completed (Fiscal Year 2020)
• Budget Amount: ¥17,420,000 (Direct Cost: ¥13,400,000、Indirect Cost: ¥4,020,000); Fiscal Year 2020: ¥5,070,000 (Direct Cost: ¥3,900,000、Indirect Cost: ¥1,170,000); Fiscal Year 2019: ¥5,070,000 (Direct Cost: ¥3,900,000、Indirect Cost: ¥1,170,000); Fiscal Year 2018: ¥7,280,000 (Direct Cost: ¥5,600,000、Indirect Cost: ¥1,680,000)
• Keywords: シナプス / 神経伝達物質 / エキソサイトーシス / エンドサイトーシス / ライブイメージング / 蛍光タンパク質 / 脳・神経 / 蛍光ライブイメージング / 神経科学 / 生理学

Outline of Final Research Achievements

Exocytosis and endocytosis of a synaptic vesicle protein in and around a presynaptic active zone were studied by live-cell fluorescence imaging of synaptophysin (syp) tagged with a pH-sensitive fluorescent protein super-ecliptic phluorin (SEP) using a novel visualization technique. Syp-SEP which had moved to the cell membrane diffused after the exocytosis. Synchronous exocytosis occurring immediately after the electrical stimulation took place at several sites within an active zone, whereas asynchronous exocytosis occurring tens to hundreds milliseconds after the stimulation took a place at different sites. We also recorded endocytosis of syp-SEP occurring in the periphery of active zone. The endocytosis occurring within a second after the stimulation and that occurring later showed different characteristics such as temperature-dependence.

Academic Significance and Societal Importance of the Research Achievements

本研究では、未解明の問題が多く残されているシナプス前部からのシナプス小胞の開口放出と、シナプス小胞膜タンパク質の細胞内への回収過程の詳細を明らかにすることに寄与する実験方法を確立し、それを活用することにより新知見を得た。これらの研究成果は、脳・神経系における情報伝達の分子・細胞機構の解明に貢献するとともに、将来的には幾多の神経・精神疾患の発症メカニズムの理解向上を介して、それらの予防・治療方法の改善に寄与することが期待できる。

Research Products

• [Journal Article] Norepinephrine facilitates induction of long-term depression through β-adrenergic receptor at parallel fiber-to-Purkinje cell synapses in the flocculus. (2020)
  • Author(s): Inoshita T, Hirano T.
  • Journal Title: Neuroscience Volume: - Pages: 141-150
  • DOI: 10.1016/j.neuroscience.2020.05.037
  • Peer Reviewed / Open Access
• [Journal Article] Amyloid-β oligomers suppress subunit-specific glutamate receptor increase during LTP (2019)
  • Author(s): Hiroshi Tanaka, Daiki Sakaguchi, Tomoo Hirano
  • Journal Title: Alzheimer’s & Dementia: Translational Research & Clinical Interventions Volume: 5 Issue: 1 Pages: 797-808
  • DOI: 10.1016/j.trci.2019.10.003
  • NAID: 120007006206
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] LTDとrebound potentiationのOKR制御における役割 (2019)
  • Author(s): 井下拓真、平野丈夫
  • Journal Title: Clinical Neuroscience Volume: 37 Pages: 940-942
• [Journal Article] Visualization of Synchronous or Asynchronous Release of Single Synaptic Vesicle in Active-Zone-Like Membrane Formed on Neuroligin-Coated Glass Surface (2018)
  • Author(s): Junichiro Funahashi, Hiromitsu Tanaka, Tomoo Hirano
  • Journal Title: Frontiers in Cellular Neuroscience Volume: 12 Pages: 1-10
  • DOI: 10.3389/fncel.2018.00140
  • NAID: 120006478048
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Suppression of AMPA Receptor Exocytosis Contributes to Hippocampal LTD (2018)
  • Author(s): Shumpei Fujii, Hiromitsu Tanaka, Tomoo Hirano
  • Journal Title: Journal of Neuroscience Volume: 38 Issue: 24 Pages: 5523-5537
  • DOI: 10.1523/jneurosci.3210-17.2018
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Synaptic m6A Epitranscriptome Reveals Functional Partitioning of Localized Transcripts for Dynamic Tripartite Synapse Modulation. (2018)
  • Author(s): Merkurjev, D., Hong, W. T., Iida, K., Oomoto, I., Goldie, B. J., Yamaguti, H., Ohara, T. Kawaguchi, S. Y., Hirano, T., Martin, K. C., Pellegrini, M. & Wang, D. O.
  • Journal Title: Nature Neuroscience Volume: 21 Issue: 7 Pages: 1004-1014
  • DOI: 10.1038/s41593-018-0173-6
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Ulk2 controls cortical excitatory?inhibitory balance via autophagic regulation of p62 and GABAA receptor trafficking in pyramidal neurons (2018)
  • Author(s): Sumitomo Akiko、Yukitake Hiroshi、Hirai Kazuko、Horike Kouta、Ueta Keisho、Chung Youjin、Warabi Eiji、Yanagawa Toru、Kitaoka Shiho、Furuyashiki Tomoyuki、Narumiya Shuh、Hirano Tomoo、Niwa Minae、Sibille Etienne、Hikida Takatoshi、Sakurai Takeshi、Ishizuka Koko、Sawa Akira、Tomoda Toshifumi
  • Journal Title: Human Molecular Genetics Volume: 27 Issue: 18 Pages: 3165-3176
  • DOI: 10.1093/hmg/ddy219
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Visualization of exo- and endocytosis of AMPA receptors during hippocampal synaptic plasticity around postsynaptic-like membrane formed on glass surface. (2018)
  • Author(s): Hirano, T.
  • Journal Title: Frontiers in Cellular Neuroscience. Volume: 12 Pages: 1-10
  • DOI: 10.3389/fncel.2018.00442
  • NAID: 120006547374
  • Peer Reviewed / Open Access
• [Journal Article] Regulation and interaction of multiple types of synaptic plasticity in a Purkinje neuron and their contribution to motor learning. (2018)
  • Author(s): Hirano,T.
  • Journal Title: Cerebellum Volume: 17 Issue: 6 Pages: 756-765
  • DOI: 10.1007/s12311-018-0963-0
  • Peer Reviewed
• [Journal Article] Purkinje Neurons: Development, Morphology and Function. (2018)
  • Author(s): Hirano,T.
  • Journal Title: Cerebellum Volume: 17 Issue: 6 Pages: 699-700
  • DOI: 10.1007/s12311-018-0985-7
  • Peer Reviewed
• [Presentation] 膜電位感受性蛍光プローブを用いた神経活動の時空間解析 (2020)
  • Author(s): 森田雅登、平野丈夫、川口真也
  • Organizer: 第72回日本細胞生物学会大会
• [Presentation] Spatio-temporal integration of synaptic inputs in hippocampal neurons studied by membrane potential imaging (2020)
  • Author(s): 森田雅登、平野丈夫、川口真也
  • Organizer: 第43回日本神経科学大会
• [Presentation] 海馬神経細胞におけるシナプス入力統合の膜電位イメージングによる時空間解析 (2020)
  • Author(s): 森田雅登、平野丈夫、川口真也
  • Organizer: 第97回日本生理学会大会
• [Presentation] Effects of β-adrenergic receptor on long-term depression at parallel fiber to Purkinje neuron synapses in the cerebellar flocculus (2019)
  • Author(s): Inoshita T, Hirano T.
  • Organizer: Neuroscience 2019, Chicago USA
  • Int'l Joint Research
• [Presentation] Different effects of noradrenaline on synaptic transmission and long-term depression at parallel fiber to Purkinje neuron synapses among cerebellar areas (2019)
  • Author(s): Inoshita T, Hirano T.
  • Organizer: 第42回 日本神経科学大会
• [Presentation] Contribution of excitatory and inhibitory synaptic plasticity in a Purkinje neuron to oculomotor learning paradigms (2018)
  • Author(s): Hirano, T.
  • Organizer: 75th Fujihara Seminar “Cerebellum as a CNS hub
  • Int'l Joint Research / Invited
• [Presentation] Subunit-specific effects on AMPA-type glutamate receptors caused by amyloid beta oligomers during hippocampal long-term potentiation (2018)
  • Author(s): Tanaka, H., Sakaguchi, D. & Hirano, T.
  • Organizer: Neuroscience 2018 in USA
  • Int'l Joint Research
• [Presentation] アミロイドベータによるサブユニット特異的なグルタミン酸受容体の動態異常 (2018)
  • Author(s): 田中 洋光, 坂口 大輝, 平野 丈夫
  • Organizer: 第41回日本分子生物学会年会
• [Presentation] Regulation of reflex eye movement and its adaptation by norepinephrine in the cerebellar flocculus. (2018)
  • Author(s): Inoshita, T. & Hirano, T.
  • Organizer: 第41回 日本神経科学大会
• [Book] 何のための脳？ AI時代の行動選択と神経科学 (2019)
  • Author(s): 平野丈夫
  • Total Pages: 145
  • Publisher: 京都大学学術出版会
  • ISBN: 9784814002306
• [Remarks] 京都大学大学院理学研究科生物科学専攻生物物理学教室神経生物学分科
  • URL: http://www.nb.biophys.kyoto-u.ac.jp/
• [Remarks] 平野研究室
  • URL: http://www.neurosci.biophys.kyoto-u.ac.jp/main.html