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Development and application of photoactivatable molecules for single-pathway activation

Research Project

Project/Area Number 18K19382
Research Category

Grant-in-Aid for Challenging Research (Exploratory)

Allocation TypeMulti-year Fund
Review Section Medium-sized Section 46:Neuroscience and related fields
Research InstitutionNational Institute for Physiological Sciences

Principal Investigator

Murakoshi Hideji  生理学研究所, 脳機能計測・支援センター, 准教授 (90608142)

Project Period (FY) 2018-06-29 – 2020-03-31
Project Status Completed (Fiscal Year 2019)
Budget Amount *help
¥6,240,000 (Direct Cost: ¥4,800,000、Indirect Cost: ¥1,440,000)
Fiscal Year 2019: ¥3,120,000 (Direct Cost: ¥2,400,000、Indirect Cost: ¥720,000)
Fiscal Year 2018: ¥3,120,000 (Direct Cost: ¥2,400,000、Indirect Cost: ¥720,000)
Keywordsシグナル伝達 / 光応答性分子
Outline of Final Research Achievements

In this project, we aimed to establish a single-pathway imaging method by combining an optogenetic tool and fluorescence imaging techniques. We have developed a new photoactivatable kinase tool that is capable of selectively activating signaling pathways. Using two-photon excitation, we successfully activated the photoactivatable kinase. Furthermore, we combined the photoactivatable kinase with 2-photon fluorescence lifetime imaging microscopy, and successfully observed the photoactivatable kinase-dependent downstream protein activity at the level of single-synapses.

Academic Significance and Societal Importance of the Research Achievements

本研究では、新規の遺伝子コード型光応答性分子の開発に成功し、単一シナプスレベルで可塑的変化を惹起できた点で学術的意義が高い。さらにこの分子を2光子蛍光寿命イメージング顕微鏡システムを用いたFRETイメージングと組み合わせた。これにより、単一経路の分子活性化イメージングが組織深部でしかも単一シナプスレベルで可能になり、新たな分子イメージング法の開拓に成功した。

Report

(3 results)
  • 2019 Annual Research Report   Final Research Report ( PDF )
  • 2018 Research-status Report
  • Research Products

    (17 results)

All 2020 2019 2018 Other

All Journal Article (6 results) (of which Int'l Joint Research: 2 results,  Peer Reviewed: 3 results,  Open Access: 3 results) Presentation (9 results) (of which Int'l Joint Research: 5 results,  Invited: 9 results) Remarks (2 results)

  • [Journal Article] 二光子蛍光寿命イメージングと色素タンパク質によるタンパク質間相互作用の可視化2020

    • Author(s)
      村越秀治、植田大海
    • Journal Title

      光学

      Volume: 49 Pages: 20-25

    • Related Report
      2019 Annual Research Report
  • [Journal Article] Reciprocal Activation within a Kinase-Effector Complex Underlying Persistence of Structural LTP.2019

    • Author(s)
      Saneyoshi T, Matsuno H, Suzuki A, Murakoshi H, Hedrick NG, Agnello E, O'Connell R, Stratton MM, Yasuda R, and Hayashi Y
    • Journal Title

      Neuron

      Volume: 102 Issue: 6 Pages: 11991210-11991210

    • DOI

      10.1016/j.neuron.2019.04.012

    • Related Report
      2019 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] ShadowR: a novel chromoprotein with reduced non-specific binding and improved expression in living cells2019

    • Author(s)
      Murakoshi Hideji、Horiuchi Hiroshi、Kosugi Takahiro、Onda Maki、Sato Aiko、Koga Nobuyasu、Nabekura Junichi
    • Journal Title

      Scientific Reports

      Volume: 9 Issue: 1 Pages: 12072-12072

    • DOI

      10.1038/s41598-019-48604-4

    • Related Report
      2019 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Rap2 and TNIK control Plexin-dependent tiled synaptic innervation in C. elegans.2018

    • Author(s)
      Chen Xi, Shibata AC, Hendi A, Kurashina M., Fortes E, Weilinger N, MacVicar B, Murakoshi H, Mizumoto K.
    • Journal Title

      eLife

      Volume: 7

    • DOI

      10.7554/elife.38801

    • Related Report
      2018 Research-status Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Neuronal Synaptic Connections Organized by Small Numbers of Molecules.2018

    • Author(s)
      Murakoshi H.
    • Journal Title

      Minorities and Small Numbers from Molecules to Organisms in Biology. Springer Singapore.

      Volume: - Pages: 7-13

    • DOI

      10.1007/978-981-13-2083-5_2

    • ISBN
      9789811320828, 9789811320835
    • Related Report
      2018 Research-status Report
  • [Journal Article] 共焦点レーザー走査顕微鏡vi.FLIM-②タイムドメイン2018

    • Author(s)
      村越秀治
    • Journal Title

      実験医学増刊 羊土社

      Volume: - Pages: 57-58

    • Related Report
      2018 Research-status Report
  • [Presentation] Optogenetic manipulation and imaging of signaling molecules in dendritic spines of neurons2020

    • Author(s)
      Hideji Murakoshi
    • Organizer
      UK-Japan Neuroscience Symposium
    • Related Report
      2019 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] Optical manipulation and imaging of signaling molecules in dendritic spines of neurons2019

    • Author(s)
      村越秀治
    • Organizer
      第60回日本組織細胞化学会総会・学術集会
    • Related Report
      2019 Annual Research Report
    • Invited
  • [Presentation] シナプス内シグナル分子の光操作と分子活性イメージング2019

    • Author(s)
      村越秀治
    • Organizer
      "Chemistry for Neuroscience 2019”研究会
    • Related Report
      2019 Annual Research Report
    • Invited
  • [Presentation] Optical manipulation and imaging of signaling molecules in dendritic spines of neurons.2019

    • Author(s)
      Hideji Murakoshi
    • Organizer
      Current Trends and Future Directions of Synapse-Circuit Plasticity Research
    • Related Report
      2019 Annual Research Report
    • Invited
  • [Presentation] Optogenetic induction of synaptic plasticity at single synapses by photoactivatable CaMKII2019

    • Author(s)
      Hideji Murakoshi
    • Organizer
      Cold Spring Harbor Laboratory Meeting
    • Related Report
      2019 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] Spatiotemporal manipulation and imaging of signaling molecules in neurons2019

    • Author(s)
      Hideji Murakoshi
    • Organizer
      The 50th NIPS International Symposium
    • Related Report
      2019 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] Optogenetic manipulation of CaMKII activity in synapses.2019

    • Author(s)
      Hideji Murakoshi
    • Organizer
      The 16th International Membrane Research Forum
    • Related Report
      2018 Research-status Report
    • Int'l Joint Research / Invited
  • [Presentation] Optical manipulation and imaging of signaling molecules in dendritic spines of neurons2018

    • Author(s)
      村越秀治
    • 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学会
    • Related Report
      2018 Research-status Report
    • Invited
  • [Presentation] Imaging Protein Activity by 2-photon Fluorescence Lifetime Imaging Microscopy2018

    • Author(s)
      村越秀治
    • Organizer
      The 66th NIBB Conference
    • Related Report
      2018 Research-status Report
    • Int'l Joint Research / Invited
  • [Remarks] 生理学研究所 村越研究室

    • URL

      http://www.nips.ac.jp/multiphoton/

    • Related Report
      2019 Annual Research Report
  • [Remarks] 村越研究室

    • URL

      http://www.nips.ac.jp/multiphoton/

    • Related Report
      2018 Research-status Report

URL: 

Published: 2018-07-25   Modified: 2023-03-16  

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