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Engineering of channelrhodopsins with improved ion selectivity as next-generation optogenetic tools and their applications

Research Project

Project/Area Number 25290002
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypePartial Multi-year Fund
Section一般
Research Field Neurophysiology / General neuroscience
Research InstitutionTohoku University

Principal Investigator

ISHIZUKA Toru  東北大学, 生命科学研究科, 講師 (10344714)

Co-Investigator(Renkei-kenkyūsha) YAWO Hiromu  東北大学, 大学院生命科学研究科, 教授 (00144353)
Research Collaborator HOSOSHIMA Shoko  
WATANABE Shota  
ZAMANI Alemeh  
HOQUE Mohammad Razuanul  
Project Period (FY) 2013-04-01 – 2016-03-31
Project Status Completed (Fiscal Year 2015)
Budget Amount *help
¥15,600,000 (Direct Cost: ¥12,000,000、Indirect Cost: ¥3,600,000)
Fiscal Year 2015: ¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000)
Fiscal Year 2014: ¥5,460,000 (Direct Cost: ¥4,200,000、Indirect Cost: ¥1,260,000)
Fiscal Year 2013: ¥6,240,000 (Direct Cost: ¥4,800,000、Indirect Cost: ¥1,440,000)
Keywordsオプトジェネティクス / チャネルロドプシン / 光遺伝学 / 微生物型ロドプシン / ナトリウムイオン / イオンチャネル
Outline of Final Research Achievements

The five glutamate residues in the second transmembrane helix of channelrhodopsin-2 (ChR2) are conserved among various ChRs. One of them, E97, interacts with cations to facilitate their permeation and is also a primary binding site for Gd(3+). However, the counterpart of MChR1 is alanine. We investigated the ion flux and the Gd(3+)-dependent channel block of MChR1. We found that the high-affinity binding site for Gd(3+) was absent, but was dependent on the negativity. The ion flux was markedly interfered with a negative charge at the position. Based on these findings, it is suggested that the channel pore of MChR1 is differently organized from that of ChR2.
Krokinobacter rhodopsin 2 (KR2) is a light-driven Na(+) pump. We introduced a codon-optimized KR2 fused with eYFP and membrane trafficking signals into cultured cortical neurons. The generation of action potential was completely blocked while KR2 activation with green light. KR2 would be useful as a next-generation optogenetic tool.

Report

(4 results)
  • 2015 Annual Research Report   Final Research Report ( PDF )
  • 2014 Annual Research Report
  • 2013 Annual Research Report
  • Research Products

    (18 results)

All 2016 2015 2014 Other

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

  • [Int'l Joint Research] フンボルト大学ベルリン(ドイツ)

    • Related Report
      2015 Annual Research Report
  • [Journal Article] The regulatory mechanism of ion permeation through a channelrhodopsin derived from Mesostigma viride (MvChR1)2016

    • Author(s)
      Watanabe S, Ishizuka T, Hososhima S, Zamani A, Hoque MR, Yawo H
    • Journal Title

      Photochemical & Photobiological Sciences

      Volume: 15 Issue: 3 Pages: 365-374

    • DOI

      10.1039/c5pp00290g

    • Related Report
      2015 Annual Research Report
    • Peer Reviewed / Open Access / Acknowledgement Compliant
  • [Journal Article] Near-infrared (NIR) up-conversion optogenetics.2015

    • Author(s)
      Shoko Hososhima, Hideya Yuasa, Toru Ishizuka, Mohammad R. Hoque, Takayuki Yamashita, Akihiro Yamanaka, Eriko Sugano, Hiroshi Tomita, and Hiromu Yawo
    • Journal Title

      Scientific Reports

      Volume: 5 Issue: 1 Pages: 16533-16533

    • DOI

      10.1038/srep16533

    • NAID

      120007179836

    • Related Report
      2015 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Membrane depolarization regulates intracellular RANKL transport in non-excitable osteoblasts2015

    • Author(s)
      Notomi T, Kuno M, Ezura Y, Honma M, Ishizuka T, Ohura K, Yao H and Noda M.
    • Journal Title

      Bone

      Volume: 81 Pages: 306-314

    • DOI

      10.1016/j.bone.2015.07.031

    • Related Report
      2015 Annual Research Report
    • Peer Reviewed / Open Access / Acknowledgement Compliant
  • [Journal Article] Structural basis for Na+ transport mechanism by a light-driven Na+ pump2015

    • Author(s)
      H. E. Kato, K. Inoue, R. Abe-Yoshizumi, Y. Kato, H. Ono, M. Konno, S. Hososhima, T. Ishizuka, M. R. Hoque, H. Kunitomo, J. Ito, S. Yoshizawa, K. Yamashita, M. Takemoto, T. Nishizawa, R. Taniguchi, K. Kogure, A. D. Maturana, Y. Iino, H. Yawo, R. Ishitani, H. Kandori*, O. Nureki*
    • Journal Title

      Nature

      Volume: 未定 Issue: 7550 Pages: 48-53

    • DOI

      10.1038/nature14322

    • Related Report
      2015 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Chimeras of Channelrhodopsin-1 and -2 from Chlamydomonas reinhardtii Exhibit Distinctive Light-induced Structural Changes from Channelrhodopsin-2.2015

    • Author(s)
      Asumi Inaguma, Hisao Tsukamoto, Hideaki E. Kato, Tetsunari Kimura, Toru Ishizuka, Satomi Oishi, Hiromu Yawo, Osamu Nureki, Yuji Furutani
    • Journal Title

      J. Biol. Chem.

      Volume: 290 Issue: 18 Pages: 11623-11634

    • DOI

      10.1074/jbc.m115.642256

    • Related Report
      2015 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Kinetic evaluation of photosensitivity in bi-stable variants of chimeric channelrhodopsins.2015

    • Author(s)
      Hososhima S, Sakai S, Ishizuka T, Yawo H.
    • Journal Title

      PLOS ONE

      Volume: - Issue: 3 Pages: e0119558-e0119558

    • DOI

      10.1371/journal.pone.0119558

    • Related Report
      2015 Annual Research Report 2014 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Engineering Biological Systems for Light2015

    • Author(s)
      石塚徹、江川遼、梅田桂子、東海林亙、八尾寛
    • Journal Title

      Seibutsu Butsuri

      Volume: 55 Issue: 6 Pages: 311-316

    • DOI

      10.2142/biophys.55.311

    • NAID

      130005110040

    • ISSN
      0582-4052, 1347-4219
    • Related Report
      2015 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Optogenetic induction of contractile ability in immature C2C12 myotubes.2015

    • Author(s)
      Asano T, Ishizuka T, Morishima K, Yawo H
    • Journal Title

      Sci Rep.

      Volume: 5 Issue: 1 Pages: 8317-8317

    • DOI

      10.1038/srep08317

    • Related Report
      2014 Annual Research Report
    • Peer Reviewed / Open Access / Acknowledgement Compliant
  • [Presentation] 近赤外オプトジェネティクスを用いたニューロンの制御2016

    • Author(s)
      細島頌子,湯浅英哉,菅野江里子,冨田浩史,石塚徹,八尾寛
    • Organizer
      第93回日本生理学会大会
    • Place of Presentation
      札幌コンベンションセンター(北海道札幌市)
    • Year and Date
      2016-03-22
    • Related Report
      2015 Annual Research Report
  • [Presentation] Optogenetic silencing of neuronal activity using a light-driven sodium ion pump in marine bacteria2015

    • Author(s)
      Ishizuka T, Hososhima S, Hoque MR, Yoshida K, Inoue K, Kandori H, Yawo H
    • Organizer
      The International Chemical Congress of Pacific Basin Societies 2015 (Pacifichem 2015)
    • Place of Presentation
      ホノルル(アメリカ合衆国)
    • Year and Date
      2015-12-15
    • Related Report
      2015 Annual Research Report
    • Int'l Joint Research
  • [Presentation] Optogenetic induction of contractile ability in C2C12 myotubes2015

    • Author(s)
      Yawo H, Asano T, Ishizuka T
    • Organizer
      8th Federation of Asian and Oceanian Physiological Societies (FAOPS) Congress
    • Place of Presentation
      バンコク(タイ)
    • Year and Date
      2015-11-22
    • Related Report
      2015 Annual Research Report
    • Int'l Joint Research
  • [Presentation] ランタニドナノ粒子のアップコンバージョンを利用した近赤外オプトジェネティクス2015

    • Author(s)
      八尾寛,細島頌子,阿部健太,湯浅英哉,菅野江里子,冨田浩史,石塚徹
    • Organizer
      第38回日本神経科学大会
    • Place of Presentation
      神戸コンベンションセンター(兵庫県神戸市)
    • Year and Date
      2015-07-28
    • Related Report
      2015 Annual Research Report
  • [Presentation] Optogenetic silencing of neural activity using a light-driven Na+ pump from marine bacteria2015

    • Author(s)
      Hoque MR,細島頌子,吉田一帆,石塚徹,井上圭一,神取秀樹,八尾寛
    • Organizer
      第38回日本神経科学大会
    • Place of Presentation
      神戸コンベンションセンター(兵庫県神戸市)
    • Year and Date
      2015-07-28
    • Related Report
      2015 Annual Research Report
  • [Presentation] Bi-stable variants of chimeric channelrhodopsins: kinetics-dependent activation of neurons2014

    • Author(s)
      Hososhima, S., Ishizuka, T., Yawo, H.
    • Organizer
      16th International Conference on Retinal Proteins
    • Place of Presentation
      長浜ロイヤルホテル(滋賀県長浜市)
    • Year and Date
      2014-10-05 – 2014-10-10
    • Related Report
      2014 Annual Research Report
  • [Presentation] Light-induced membrane depolarization by two-component optogenetics2014

    • Author(s)
      Zamani, A., Ishizuka, T., Yawo, H.
    • Organizer
      16th International Conference on Retinal Proteins
    • Place of Presentation
      長浜ロイヤルホテル(滋賀県長浜市)
    • Year and Date
      2014-10-05 – 2014-10-10
    • Related Report
      2014 Annual Research Report
  • [Presentation] Modification of ion selectivity in channelrhodopsins2014

    • Author(s)
      Ishizuka, T.
    • Organizer
      Channelrhodopsin et al. Optogenetic Tools and Applications, 2014 Conference
    • Place of Presentation
      ビュルツブルク(ドイツ)
    • Year and Date
      2014-09-28 – 2014-10-01
    • Related Report
      2014 Annual Research Report
  • [Book] 光と生命の事典2016

    • Author(s)
      日本光生物学協会 光と生命の事典 編集委員会編 編集委員-真嶋哲朗,飯野盛利,七田芳則,藤堂剛,執筆者-石塚徹,他
    • Total Pages
      436
    • Publisher
      朝倉書店
    • Related Report
      2015 Annual Research Report

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Published: 2013-05-21   Modified: 2019-07-29  

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