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Application of fluorescent probes to single vesicle analysis

Research Project

Project/Area Number 15K14321
Research Category

Grant-in-Aid for Challenging Exploratory Research

Allocation TypeMulti-year Fund
Research Field Neurophysiology / General neuroscience
Research InstitutionDoshisha University

Principal Investigator

Sakaba Takeshi  同志社大学, 脳科学研究科, 教授 (80609511)

Co-Investigator(Renkei-kenkyūsha) KAWAGUCHI Shinya  京都大学, 産官学連携本部, 特定准教授 (00378530)
MIDORIKAWA Mitsuharu  東京女子医科大学, 医学部, 准講師 (60632643)
Project Period (FY) 2015-04-01 – 2018-03-31
Project Status Completed (Fiscal Year 2017)
Budget Amount *help
¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000)
Fiscal Year 2017: ¥650,000 (Direct Cost: ¥500,000、Indirect Cost: ¥150,000)
Fiscal Year 2016: ¥650,000 (Direct Cost: ¥500,000、Indirect Cost: ¥150,000)
Fiscal Year 2015: ¥2,600,000 (Direct Cost: ¥2,000,000、Indirect Cost: ¥600,000)
Keywordsシナプス / 神経 / イメージング / 神経科学 / 生理学
Outline of Final Research Achievements

In mammalian CNS synapses, transmitter is released by fusion of synaptic vesicles to the plasma membrane. Because diameter of synaptic vesicles is small, it is difficult to look at dynamics of single synaptic vesicles. In this program, we have applied total internal reflection fluorescent microscopy to acutely-dissociated presynaptic terminals. By sparsely staining synaptic vesicles with FM1-43, we could monitor dynamics of synaptic vesicles such as fusion and tethering to the release sites at the calyx of Held synapse in the auditory brainstem as well as hippocampal mossy fiber synapse. Moreover, we found that tethering of synaptic vesicles to the release sites was not sufficient for vesicle fusion. In addition to tethering, molecular priming of synaptic vesicles was required for fusion events.

Report

(4 results)
  • 2017 Annual Research Report   Final Research Report ( PDF )
  • 2016 Research-status Report
  • 2015 Research-status Report
  • Research Products

    (16 results)

All 2018 2017 2016 2015 Other

All Int'l Joint Research (1 results) Journal Article (7 results) (of which Int'l Joint Research: 2 results,  Peer Reviewed: 7 results,  Open Access: 2 results,  Acknowledgement Compliant: 4 results) Presentation (6 results) (of which Int'l Joint Research: 1 results,  Invited: 4 results) Remarks (2 results)

  • [Int'l Joint Research] マックスプランク実験医学研究所(ドイツ)

    • Related Report
      2016 Research-status Report
  • [Journal Article] Kinetics of transmitter release at the calyx of Held synapse2018

    • Author(s)
      Sakaba T.
    • Journal Title

      Proceedings of the Japan Academy, Series B

      Volume: 94 Issue: 3 Pages: 139-152

    • DOI

      10.2183/pjab.94.010

    • NAID

      130006464084

    • ISSN
      0386-2208, 1349-2896
    • Related Report
      2017 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Kinetics of Releasable Synaptic Vesicles and Their Plastic Changes at Hippocampal Mossy Fiber Synapses.2017

    • Author(s)
      Midorikawa M, Sakaba T.
    • Journal Title

      Neuron

      Volume: 96 Issue: 5 Pages: 1033-1040

    • DOI

      10.1016/j.neuron.2017.10.016

    • Related Report
      2017 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Fast Ca2+ Buffer-Dependent Reliable but Plastic Transmission at Small CNS Synapses Revealed by Direct Bouton Recording.2017

    • Author(s)
      Kawaguchi SY, Sakaba T.
    • Journal Title

      Cell Reports

      Volume: 21 Issue: 12 Pages: 3338-3345

    • DOI

      10.1016/j.celrep.2017.11.072

    • Related Report
      2017 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Synaptic Vesicle Endocytosis Occurs on Multiple Timescales and Is Mediated by Formin-Dependent Actin Assembly.2017

    • Author(s)
      Soykan T, Kaempf N, Sakaba T, Vollweiter D, Goerdeler F, Puchkov D, Kononenko NL, Haucke V.
    • Journal Title

      Neuron

      Volume: 93 Issue: 4 Pages: 854-866

    • DOI

      10.1016/j.neuron.2017.02.011

    • Related Report
      2016 Research-status Report
    • Peer Reviewed / Int'l Joint Research / Acknowledgement Compliant
  • [Journal Article] Distinct modes of endocytotic presynaptic membrane and protein uptake at the calyx of Held terminal of rats and mice.2016

    • Author(s)
      Okamoto Y, Lipstein N, Hua Y, Lin KH, Brose N, Sakaba T, Midorikawa M.
    • Journal Title

      eLife

      Volume: 5

    • DOI

      10.7554/elife.14643

    • Related Report
      2016 Research-status Report
    • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
  • [Journal Article] Ca(2+) current facilitation determines short-term facilitation at inhibitory synapses between cerebellar Purkinje cells.2015

    • Author(s)
      Francois Diaz-Rojas, Takeshi Sakaba, Shin-ya Kawaguchi
    • Journal Title

      The Journal of Physiology

      Volume: 593 Issue: 22 Pages: 4889-4904

    • DOI

      10.1113/jp270704

    • Related Report
      2015 Research-status Report
    • Peer Reviewed / Acknowledgement Compliant
  • [Journal Article] Imaging Exocytosis of Single Synaptic Vesicles at a Fast CNS Presynaptic Terminal.2015

    • Author(s)
      Midorikawa M, Sakaba T.
    • Journal Title

      Neuron

      Volume: 88 Issue: 3 Pages: 492-8

    • DOI

      10.1016/j.neuron.2015.09.047

    • Related Report
      2015 Research-status Report
    • Peer Reviewed / Acknowledgement Compliant
  • [Presentation] 哺乳類シナプス前終末の生理学的研究2017

    • Author(s)
      坂場 武史
    • Organizer
      日本生理学会
    • Place of Presentation
      浜松アクトシティ(静岡県・浜松市)
    • Year and Date
      2017-03-28
    • Related Report
      2016 Research-status Report
  • [Presentation] Mechansims of transmitter release at the calyx of Held synapse..2017

    • Author(s)
      Sakaba T
    • Organizer
      Goettingen Neurocience meeting
    • Place of Presentation
      ゲッチンゲン(ドイツ)
    • Year and Date
      2017-03-23
    • Related Report
      2016 Research-status Report
  • [Presentation] パッチクランプ法をはじめとする電気生理学的計測法について2017

    • Author(s)
      坂場 武史
    • Organizer
      日本神経科学学会
    • Related Report
      2017 Annual Research Report
    • Invited
  • [Presentation] 中枢神経系におけるシナプス伝達の生理学、塚原仲晃賞記念講演2016

    • Author(s)
      坂場 武史
    • Organizer
      日本神経科学学会
    • Place of Presentation
      パシフィコ横浜(神奈川県・横浜市)
    • Year and Date
      2016-07-20
    • Related Report
      2016 Research-status Report
    • Invited
  • [Presentation] Physiology of axon and axon terminals.2015

    • Author(s)
      Sakaba, T
    • Organizer
      The 6th Faons congress and the 11th Biennial conference of CNS
    • Place of Presentation
      Wuzhen, Zhejiang Province, China
    • Year and Date
      2015-09-21
    • Related Report
      2015 Research-status Report
    • Int'l Joint Research / Invited
  • [Presentation] シナプス前終末の生理学2015

    • Author(s)
      坂場 武史
    • Organizer
      日本糖質学会
    • Place of Presentation
      東京大学 (東京)
    • Year and Date
      2015-08-02
    • Related Report
      2015 Research-status Report
    • Invited
  • [Remarks] 同志社大学大学院脳科学研究科

    • URL

      http://brainscience.doshisha.ac.jp/introduction/mol/msf.html

    • Related Report
      2017 Annual Research Report
  • [Remarks] 同志社大学大学院脳科学研究科シナプス分子機能部門

    • URL

      http://brainscience.doshisha.ac.jp/introduction/mol/msf.html

    • Related Report
      2015 Research-status Report

URL: 

Published: 2015-04-16   Modified: 2019-03-29  

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