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Development and application of multiphoton excitation for simultaneous multi-color measurement and manipulation of neural activity

Research Project

Project/Area Number 21K19429
Research Category

Grant-in-Aid for Challenging Research (Exploratory)

Allocation TypeMulti-year Fund
Review Section Medium-sized Section 51:Brain sciences and related fields
Research InstitutionKyoto University

Principal Investigator

Sakamoto Masayuki  京都大学, 生命科学研究科, 特定准教授 (00777865)

Project Period (FY) 2021-07-09 – 2023-03-31
Project Status Completed (Fiscal Year 2022)
Budget Amount *help
¥6,500,000 (Direct Cost: ¥5,000,000、Indirect Cost: ¥1,500,000)
Fiscal Year 2022: ¥3,250,000 (Direct Cost: ¥2,500,000、Indirect Cost: ¥750,000)
Fiscal Year 2021: ¥3,250,000 (Direct Cost: ¥2,500,000、Indirect Cost: ¥750,000)
Keywordsカルシウムセンサー / 光遺伝学ツール / タンパク質工学 / 多色イメージング / 多光子励起 / カルシウムイメージング / 2光子励起顕微鏡 / 広視野顕微鏡 / FRET / 光遺伝学
Outline of Research at the Start

近年、カルシウムセンサーと光遺伝学ツールを用いることで、神経活動計測・操作が生体においても可能となってきている。しかしながら、従来の手法では、光遺伝学ツールがイメージングの励起光で活性化されてしまうなど、実用的な使用に向けては多くの課題が残っている。本研究では、新規光遺伝学ツールをデザインをおこない、多色イメージングと光遺伝学的操作が同時に可能な方法の開発をおこなう。

Outline of Final Research Achievements

We developed a new optogenetic tool and novel calcium indicator to establish a method for simultaneous calcium imaging and optogenetic manipulation in vivo. Through screening, we successfully developed a mutant with larger conductance than existing optogenetic tools. Furthermore, we obtained a new red calcium sensor, RCaMP3, that exhibited increased fluorescence brightness and fluorescence change rate compared to conventional red calcium indicators.

Academic Significance and Societal Importance of the Research Achievements

高次脳機能を制御する神経ネットワークを理解するためには、1細胞レベルの解像度で神経活動計測と神経活動操作を同時におこなう技術が必要である。本研究で開発した新規プローブを用いることで、神経活動計測と活動操作がクロストークフリーに生体脳で実現できるだけでなく、旧来の生理学の枠を超えた神経科学の問題に取り組みことも可能となる。

Report

(3 results)
  • 2022 Annual Research Report   Final Research Report ( PDF )
  • 2021 Research-status Report
  • Research Products

    (16 results)

All 2023 2022 2021 Other

All Int'l Joint Research (4 results) Journal Article (4 results) (of which Int'l Joint Research: 1 results,  Peer Reviewed: 3 results,  Open Access: 4 results) Presentation (8 results) (of which Int'l Joint Research: 2 results,  Invited: 8 results)

  • [Int'l Joint Research] University of Copenhagen(デンマーク)

    • Related Report
      2022 Annual Research Report
  • [Int'l Joint Research] Istituto Italiano di Tecnologia(イタリア)

    • Related Report
      2022 Annual Research Report
  • [Int'l Joint Research] Yale University/Columbia University/University of Washington(米国)

    • Related Report
      2022 Annual Research Report
  • [Int'l Joint Research] Beijing Normal University(中国)

    • Related Report
      2022 Annual Research Report
  • [Journal Article] A multicolor suite for deciphering population coding in calcium and cAMP<i>in vivo</i>2023

    • Author(s)
      Yokoyama Tatsushi、Manita Satoshi、Uwamori Hiroyuki、Tajiri Mio、Imayoshi Itaru、Yagishita Sho、Murayama Masanori、Kitamura Kazuo、Sakamoto Masayuki
    • Journal Title

      bioRxiv

      Volume: -

    • DOI

      10.1101/2023.01.06.522686

    • Related Report
      2022 Annual Research Report
    • Open Access
  • [Journal Article] In utero electroporation and cranial window implantation for in vivo wide-field two-photon calcium imaging using G-CaMP9a transgenic mice.2022

    • Author(s)
      Sakamoto M, Ota K, Kondo Y, Okamura M, Fujii H, Bito H.
    • Journal Title

      Star Protocols

      Volume: 3 Issue: 2 Pages: 101421-101421

    • DOI

      10.1016/j.xpro.2022.101421

    • Related Report
      2022 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] A Flp-dependent G-CaMP9a transgenic mouse for neuronal imaging in vivo2022

    • Author(s)
      Sakamoto Masayuki, Inoue Masatoshi, Takeuchi Atsuya, Kobari Shigetaka, Yokoyama Tatsushi, Horigane Shin-ichiro, Takemoto-Kimura Sayaka, Abe Manabu, Sakimura Kenji, Kano Masanobu, Kitamura Kazuo, Fujii Hajime, Bito Haruhiko
    • Journal Title

      Cell Reports Methods

      Volume: 2 Issue: 2 Pages: 100168-100168

    • DOI

      10.1016/j.crmeth.2022.100168

    • URL

      https://pure.teikyo.jp/en/publications/8054d3cd-fd72-478d-8c3f-48fd86436357

    • Related Report
      2021 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] Imaging Voltage with Microbial Rhodopsins2021

    • Author(s)
      Zhang Xiao Min、Yokoyama Tatsushi、Sakamoto Masayuki
    • Journal Title

      Frontiers in Molecular Biosciences

      Volume: 8 Pages: 733829-733829

    • DOI

      10.3389/fmolb.2021.738829

    • Related Report
      2021 Research-status Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Presentation] Imaging voltage and biochemical signaling in vivo2022

    • Author(s)
      Masayuki Sakamoto
    • Organizer
      Merocyanine 540/Flash meeting
    • Related Report
      2022 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] Fluorescent biosensors for probing neuronal activity and biochemical signaling2022

    • Author(s)
      Masayuki Sakamoto
    • Organizer
      Cell Biology, Developmental Biology, and Systems Biology Course Meeting
    • Related Report
      2022 Annual Research Report
    • Invited
  • [Presentation] 神経活動可視化のための蛍光プローブの開発と応用2022

    • Author(s)
      坂本雅行
    • Organizer
      第11回パラレル脳研究部門勉強会・セミナー
    • Related Report
      2022 Annual Research Report
    • Invited
  • [Presentation] Fluorescent biosensors for probing neuronal activity and biochemical signaling2022

    • Author(s)
      Masayuki Sakamoto
    • Organizer
      Hamburg University - Kyoto University Joint Symposium “Cellular mechanisms of learning and memory”
    • Related Report
      2022 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] 神経活動可視化のための蛍光プローブの開発と応用2022

    • Author(s)
      坂本雅行
    • Organizer
      第5回これからの神経回路研究の会
    • Related Report
      2022 Annual Research Report
    • Invited
  • [Presentation] Engineering microbial rhodopsins for genetically encoded voltage indicators2021

    • Author(s)
      Masayuki Sakamoto
    • Organizer
      第44回日本神経科学大会JNS-CAN合同シンポジウム「Understanding of synaptic and neural circuits through advanced optical technologies.」
    • Related Report
      2021 Research-status Report
    • Invited
  • [Presentation] 膜電位感受性プローブを用いた神経活動計測2021

    • Author(s)
      坂本雅行
    • Organizer
      第64 回日本神経化学会大会シンポジウム「先端的イメージング技術で読み解く高次脳機能の仕組み」
    • Related Report
      2021 Research-status Report
    • Invited
  • [Presentation] 膜電位感受性プローブを用いた神経活動計測2021

    • Author(s)
      坂本雅行
    • Organizer
      第42 回生体膜と薬物の相互作用シンポジウム
    • Related Report
      2021 Research-status Report
    • Invited

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Published: 2021-07-13   Modified: 2024-01-30  

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