Creation of channelrhodopsin variants which have various properties in ion selectivity and localization

• Project/Area Number: 25670103
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: General physiology
• Research Institution: Tohoku University
• Principal Investigator: YAWO Hiromu 東北大学, 生命科学研究科, 教授 (00144353)
• Project Period (FY): 2013-04-01 – 2015-03-31
• Project Status: Completed (Fiscal Year 2014)
• Budget Amount: ¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000); Fiscal Year 2014: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000); Fiscal Year 2013: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
• Keywords: 光遺伝学 / オプトジェネティクス / チャネルロドプシン / キネティクス / イオン選択性 / 近赤外 / ランタニドナノ粒子 / アップコンバージョン / イオンチャネル / Gd3+ / オルガネラ / 筋小胞体 / 筋収縮 / サルコメア / 受容体 / 国際情報交換 / アメリカ合衆国

Outline of Final Research Achievements

(1) We generated step-function opsin (SFO) variants of ChRWR and ChRFR, chimeric channelrhodopsins between ChR1 and ChR2, and analyzed kinetics of their photocurrents as functions of wavelength, fluence and irradiation time. One of them, ChRFR-C167A was expressed in the rat cortical neuron and the light-dependent change of membrane potential was quantitatively analyzed as a function of fluence and irradiation time. (2) We generated enhaced variants of MvChR1, which is more selective to Na+ than other ChRs. The mode of ion permeation was revealed to be different from other ChRs. (3) We made a near-infrared (NIR) optogenetics system which consists of lanthanide nanoparticles (LNP) as donors and ChRs as acceptors. The firing of neurons expressing ChRs were controled by NIR light through upconversion of LNPs.

Research Products

• [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
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [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
  • Peer Reviewed / Open Access
• [Journal Article] Structural basis for Na+ transport mechanism by a light-driven Na+ pump (2015)
  • 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
  • Peer Reviewed / Open Access
• [Journal Article] Chimeras of channelrhodopsin-1 and -2 from Chlamydomonas reinhardtii possess distinctive light-induced structural changes with less protonation change of a key glutamate residue for the gating behavior. (2015)
  • Author(s): Inaguma A, Tsukamoto H, Kato HE, Kimura T, Ishizuka T, Yawo H, Nureki O, Furutani Y
  • Journal Title: J Biol Chem Volume: Mar 21
  • Peer Reviewed
• [Journal Article] Near-infrared (NIR) optogenetics using up-conversion system (2015)
  • Author(s): Shoko Hososhima, Hideya Yuasa, Toru Ishizuka, Hiromu Yawo
  • Journal Title: Proc. SPIE, Optical Techniques in Neurosurgery, Neurophotonics, and Optogenetics II Volume: 9305 Pages: 93052R-93052R
  • DOI: 10.1117/12.2078875
  • Peer Reviewed
• [Journal Article] Optogenetic patterning of whisker-barrel cortical system in transgenic rat expressing channelrhodopsin-2. (2014)
  • Author(s): Honjoh T, Ji Z-G, Yokoyama Y, Sumiyoshi A, Shibuya Y, Matsuzaka Y, Kawashima R, Mushiake H, Ishizuka T, Yawo H.
  • Journal Title: PLoS ONE Volume: 9 (4) Issue: 4 Pages: 28134-28170
  • DOI: 10.1371/journal.pone.0093706
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Regulation of later neurogenic stages of adult-derived neural stem/progenitor cells by L-type Ca2+ channels. (2014)
  • Author(s): Teh DBL, Ishizuka T, Yawo H
  • Journal Title: Develop Growth Differ. Volume: 56 Pages: 583-594
  • Peer Reviewed
• [Journal Article] Photofunctional nanomodulators for bioexcitation. (2014)
  • Author(s): Miyako E, Russier J, MauroM, Cebrian C, Yawo H, Menard-Moyon C, Hutchison JA, Yudasaka M, Iijima S, De Cola L, Bianco A
  • Journal Title: Angew Chem Int Ed. Volume: 53 Issue: 48 Pages: 13121-13125
  • DOI: 10.1002/anie.201407169
  • Peer Reviewed
• [Journal Article] オプトジェネティクスによる細胞機能の光操作． (2014)
  • Author(s): 江川遼，八尾寛
  • Journal Title: 細胞 Volume: 46 Pages: 35-38
  • NAID: 40020155660
• [Journal Article] Optogenetic stimulation of the auditory pathway. (2014)
  • Author(s): Hernandez VH, Gehrt A, Reuter K, Jing Z, Jeschke M, Mendoza Schulz A, Hoch G, Bartels M, Vogt G, Garnham CW, Hiromu Yawo H, Fukazawa Y, Augustine GJ, Bamberg E, Kügler S, Salditt T, de Hoz L, Strenzke N, Moser T
  • Journal Title: J Clin Invest Volume: 124 Issue: 3 Pages: 1114-1129
  • DOI: 10.1172/jci69050
  • Peer Reviewed
• [Journal Article] 光遺伝学に有用なツール開発：分子、遺伝子導入、光照射の実際. (2014)
  • Author(s): 八尾 寛、江川 遼
  • Journal Title: 細胞工学 Volume: 33 Pages: 243-248
• [Journal Article] Optogenetically induced Seizure and the Longitudinal Hippocampal Network. (2013)
  • Author(s): Shin-ichiro Osawa, , Hajime Mushiake
  • Journal Title: PLos One Volume: 8 Issue: 4 Pages: 1-1
  • DOI: 10.1371/journal.pone.0060928
  • Peer Reviewed
• [Journal Article] Optogenetic manipulation of neural and non-neural functions. (2013)
  • Author(s): Yawo H, Asano T, Sakai S, Ishizuka T
  • Journal Title: Development, Growth & Differentiation Volume: 55 Issue: 4 Pages: 474-490
  • DOI: 10.1111/dgd.12053
  • Peer Reviewed
• [Journal Article] Hindbrain V2a neurons in the excitation of spinal locomotor circuits during zebrafish swimming (2013)
  • Author(s): Kimura, Y., Satou, C., Fujioka, S., Shoji, W., Umeda, K., Ishizuka, T., Yawo, H., and Higashijima, S.
  • Journal Title: Current Biology Volume: 23 Issue: 10 Pages: 843-849
  • DOI: 10.1016/j.cub.2013.03.066
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] オプトジェネティクスを用いたin vivoラット脳の双方向性プロービング． (2013)
  • Author(s): 本城達也，八尾 寛
  • Journal Title: 実験医学 Volume: 31 Pages: 927-933
• [Presentation] チャネルロドプシン光電変換の特性と応用 (2015)
  • Author(s): 八尾 寛
  • Organizer: 理研シンポジウム「細胞システムの動態と論理 Ⅶ」
  • Place of Presentation: 和光市
  • Year and Date: 2015-04-02 – 2015-04-03
  • Invited
• [Presentation] A new reporter rat line which express bright red fluorescent protein (tdTomato) conditionally under Cre-loxP system. (2015)
  • Author(s): Igarashi H, Koizumi K, Kaneko R, Ikeda K, Onimaru H, Yanagawa Y, Muramatsu S-I, Ishizuka T, Yawo H
  • Organizer: 第92回日本生理学会大会
  • Place of Presentation: 神戸
  • Year and Date: 2015-03-21 – 2015-03-23
• [Presentation] Discrimination of optogenetic whisker pattern in channelrhodopsin-2 transgenic rat. (2015)
  • Author(s): Abe K, Ishizuka T, Yawo H
  • Organizer: 第92回日本生理学会大会
  • Place of Presentation: 神戸
  • Year and Date: 2015-03-21 – 2015-03-23
• [Presentation] Opt-fMRI study of whisker-barrel cortical responses using channelrhodopsin-2 expressing rat. (2015)
  • Author(s): Yokoyama Y, Sumiyoshi A, Kawashima R, Yawo H
  • Organizer: 第92回日本生理学会大会
  • Place of Presentation: 神戸
  • Year and Date: 2015-03-21 – 2015-03-23
• [Presentation] チャネルロドプシンの分子ダイナミクス. (2015)
  • Author(s): 八尾寛
  • Organizer: 蛋白研セミナー「光運動反応・光センサー蛋白質・光遺伝学」
  • Place of Presentation: 吹田
  • Year and Date: 2015-03-10 – 2015-03-11
  • Invited
• [Presentation] Near-infrared (NIR) optogenetics using up-conversion system. (2015)
  • Author(s): Hososhima S, Yuasa H, Ishizuka T, Yawo H
  • Organizer: SPIE Photonics West-Bios
  • Place of Presentation: San Francisco, USA
  • Year and Date: 2015-02-07 – 2015-02-12
• [Presentation] 光で生命機能に干渉する ～オプトジェネティクス（光遺伝学）の現況と展望～（チュートリアル）. (2014)
  • Author(s): 八尾寛
  • Organizer: 応用物理学会 量子エレクトロニクス研究会
  • Place of Presentation: 軽井沢
  • Year and Date: 2014-12-19 – 2014-12-20
  • Invited
• [Presentation] 触覚時空間パターン脳内表現のオプトジェネティクス. (2014)
  • Author(s): 八尾寛
  • Organizer: 第35 回日本レーザー医学会総会
  • Place of Presentation: 東京
  • Year and Date: 2014-11-29 – 2014-11-30
  • Invited
• [Presentation] ランタニドナノ粒子のアップコンバージョン効果を応用した生体の近赤外光操作. (2014)
  • Author(s): 細島頌子, 湯浅英哉, 石塚徹, 八尾寛
  • Organizer: 第36回日本バイオマテリアル学会大会
  • Place of Presentation: 東京
  • Year and Date: 2014-11-17 – 2014-11-18
• [Presentation] Developmental connectomics of axonal reorganization in avian ciliary ganglion. (2014)
  • Author(s): R. Egawa, S. Hososhima, T. Ishizuka, H. Yawo
  • Organizer: Neuroscience 2014
  • Place of Presentation: Washington DC, USA
  • Year and Date: 2014-11-15 – 2014-11-19
• [Presentation] Evaluation of a novel reporter rat line which conditionally expresses red fluorescent protein (tdTomato). (2014)
  • Author(s): H. Igarashi, K. Koizumi, R. Kaneko, K. Ikeda, H. Onimaru, Y. Yanagawa, S.-I. Muramatsu, T. Ishizuka, H. Yawo
  • Organizer: Neuroscience 2014
  • Place of Presentation: Washington DC, USA
  • Year and Date: 2014-11-15 – 2014-11-19
• [Presentation] 感覚の遺伝子工学. (2014)
  • Author(s): 八尾寛
  • Organizer: 第５回感性センシング応用ロードマップ技術分科会
  • Place of Presentation: 東京
  • Year and Date: 2014-11-04
  • Invited
• [Presentation] Optogenetic patterning of touch perception -- sensing light by skin. (2014)
  • Author(s): Yawo H, Yokoyama Y, Sumiyoshi A, Abe A, Koizumi K, Egawa R, Liu Y, Ohshiro T, Matsuzaka Y, Kawashima R, Mushiake H, Ishizuka T.
  • Organizer: The 16th International Conference on Retinal Proteins (ICRP2014)
  • Place of Presentation: Nagahama, Japan
  • Year and Date: 2014-10-05 – 2014-10-10
  • Invited
• [Presentation] Bi-stable variants of chimeric channelrhodopsins - kinetics-dependent activation of neurons. (2014)
  • Author(s): Hososhima S, Ishizuka T, Yawo H
  • Organizer: The 16th International Conference on Retinal Proteins (ICRP2014)
  • Place of Presentation: Nagahama, Japan
  • Year and Date: 2014-10-05 – 2014-10-10
• [Presentation] Light-induced membrane depolarization by two-component optogenetic. (2014)
  • Author(s): A. Zamani, T. Ishizuka, H. Yawo
  • Organizer: The 16th International Conference on Retinal Proteins (ICRP2014)
  • Place of Presentation: Nagahama, Japan
  • Year and Date: 2014-10-05 – 2014-10-10
• [Presentation] Functional MRI signal to optogenetic tactile pattern of channelrhodopsin-2 expressing rat in barrel cortex. (2014)
  • Author(s): Y. Yokoyama, A. Sumiyoshi, R. Kawashima, H. Yawo
  • Organizer: The 16th International Conference on Retinal Proteins (ICRP2014)
  • Place of Presentation: Nagahama, Japan
  • Year and Date: 2014-10-05 – 2014-10-10
• [Presentation] 触覚パターンのオプトジェネティクス制御. (2014)
  • Author(s): 八尾寛, 横山超一, 住吉晃, 阿部健太, 小泉協, 江川遼, 劉越人, 大城朝一, 松坂義哉, 川島隆太, 虫明元, 石塚徹
  • Organizer: 第52回日本生物物理学会年会
  • Place of Presentation: 札幌
  • Year and Date: 2014-09-25 – 2014-09-27
  • Invited
• [Presentation] 赤色蛍光タンパク質tdTomatoをCre-loxPコンディショナルに発現するレポーターラットの評価． (2014)
  • Author(s): 五十嵐敬幸，小泉協，金子涼輔，池田啓子，鬼丸洋，柳川右千夫，村松 慎一，石塚 徹，八尾寛
  • Organizer: 第37回日本神経科学大会
  • Place of Presentation: 横浜
  • Year and Date: 2014-09-11 – 2014-09-13
• [Presentation] ニワトリ胚毛様体神経節の発達期におけるカスパーゼ6依存的なシナプスリモデリング． (2014)
  • Author(s): 加藤秀理，江川遼，細島頌子，石塚徹，八尾寛
  • Organizer: 第37回日本神経科学大会
  • Place of Presentation: 横浜
  • Year and Date: 2014-09-11 – 2014-09-13
• [Presentation] ニワトリ胚毛様体神経節における軸索再編成の発達期コネクトミクス研究． (2014)
  • Author(s): 江川遼，細島頌子，石塚徹，八尾寛
  • Organizer: 第37回日本神経科学大会
  • Place of Presentation: 横浜
  • Year and Date: 2014-09-11 – 2014-09-13
• [Presentation] オプトジェネティックスを用いた触覚パターンによるバレル野のfMRI応答． (2014)
  • Author(s): 横山超一，住吉晃，川島隆太，八尾寛
  • Organizer: 第37回日本神経科学大会
  • Place of Presentation: 横浜
  • Year and Date: 2014-09-11 – 2014-09-13
• [Presentation] 光を触覚として感じるラット (2014)
  • Author(s): 八尾寛
  • Organizer: 光操作研究会シンポジウム
  • Place of Presentation: 仙台
  • Year and Date: 2014-08-21 – 2014-08-22
  • Invited
• [Presentation] Target-dependent direction of Muse cell differentiation -- a proposal of in vitro model system. (2014)
  • Author(s): H. Igarashi, T. Morita, T. Murakami, Y. Matsuzaki, A. Konno, T. Ishizuka, M. Dezawa, H. Yawo
  • Organizer: 5th International Congress on Stem Cells and Tissue Formation
  • Place of Presentation: Dresden, Germany
  • Year and Date: 2014-07-08 – 2014-07-11
• [Presentation] 発達期ニワトリ胚毛様体神経節で見られるプレシナプス刈り込みへのカスパーゼ6の関与 (2014)
  • Author(s): 加藤秀理，江川遼，細島頌子，石塚徹，八尾寛
  • Organizer: 第47回発生生物学会
  • Place of Presentation: 名古屋
  • Year and Date: 2014-05-27 – 2014-05-30
• [Presentation] Rat-Based Systems for Basic and Clinical Optogenetics. (2014)
  • Author(s): Yawo H
  • Organizer: Gordon Research Conference on Photosensory Receptors and Signal Transduction
  • Place of Presentation: Lucca (Barga), Italy
  • Year and Date: 2014-04-06 – 2014-04-11
  • Invited
• [Presentation] The molecular determinants involved in the Gd3+-dependent channel block of channelrhodopsins. (2014)
  • Author(s): Yawo H, Watanabe S, Ishizuka T
  • Organizer: Gordon Research Conference on Photosensory Receptors and Signal Transduction
  • Place of Presentation: Lucca (Barga), Italy
  • Year and Date: 2014-04-06 – 2014-04-11
• [Presentation] Optogenetic Patterning of Somatosensory Perception (2014)
  • Author(s): Yawo H
  • Organizer: "International Institute for Advanced Studies" Conference
  • Place of Presentation: 木津川市
  • Invited
• [Presentation] キメラチャネルロドプシンの双安定性 (2014)
  • Author(s): 細島頌子、石塚徹、八尾 寛
  • Organizer: 第91回日本生理学会大会
  • Place of Presentation: 鹿児島
• [Presentation] Light-Evoked Touch Sense with Channelrhodopsin-Optogenetics (2013)
  • Author(s): Yawo H
  • Organizer: NIH-Tohoku University-JSPS Symposium
  • Place of Presentation: 仙台
  • Invited
• [Presentation] Optogenetic delivery of touch sense into peripheral and central nervous systems (2013)
  • Author(s): Yawo H
  • Organizer: IUPS2013
  • Place of Presentation: Birmingham, イギリス
  • Invited
• [Presentation] The molecular determinants involved in ion flux regulation of channelrhodopsins (2013)
  • Author(s): Yawo H
  • Organizer: International Symposium Optogenetics 2013
  • Place of Presentation: 東京
  • Invited
• [Presentation] Optogenetic patterning of whisker-barrel tactile perception (2013)
  • Author(s): Yawo H
  • Organizer: A symposium organized by the WCI Center for Functional Connectomics “lluminating brain function: New tools and new applications”
  • Place of Presentation: Seoul, 韓国
  • Invited
• [Presentation] Optogenetic manipulation of presynaptic activity in the ciliary ganglion during embryogenesis of the chick (2013)
  • Author(s): Shoko Hososhima, Toru lshizuka, Hiromu Yawo
  • Organizer: IUPS 2013
  • Place of Presentation: Birmingham, イギリス
• [Presentation] A DMD-based multicolor irradiation system for the optogenetic analysis of neural circuits (2013)
  • Author(s): Seiichiro Sakai, Takaaki Mishima, Toru lshizuka, Hiromu Yawo
  • Organizer: IUPS 2013
  • Place of Presentation: Birmingham, イギリス
• [Presentation] Whisker photostimulation evokes barrel cortical response of ChR2 transgenic rat (2013)
  • Author(s): Tatsuya Honjoh, Akira Sumiyoshi, Yukinobu Yokoyama, Zhi-Gang Ji, Toru lshizuka, Ryuta Kawashima, Hiromu Yawo
  • Organizer: IUPS 2013
  • Place of Presentation: Birmingham, イギリス
• [Presentation] Involvement of caspases in the synaptogenesis of chick ciliary ganglion (2013)
  • Author(s): Hidetaka Katow, Ryo Egawa, Shoko Hososhima, Toru lshizuka, Hiromu Yawo
  • Organizer: IUPS 2013
  • Place of Presentation: Birmingham, イギリス
• [Presentation] Optogenetically-evoked Ca2+ release from intracellular store in the calyx-type presynaptic terminal of the embryonic chick ciliary ganglion (2013)
  • Author(s): Ryo Egawa, Shoko Hososhima, Hidetaka Katow, Toru Ishizuka, Hiromu Yawo
  • Organizer: IUPS 2013
  • Place of Presentation: Birmingham, イギリス
• [Presentation] 光遺伝学的に誘起された杯状シナプス前終末小胞体ストアからのCa2+放出 (2013)
  • Author(s): 江川遼、細島頌子、加藤秀理、石塚徹、八尾寛
  • Organizer: 第45回東北生理談話会
  • Place of Presentation: 仙台
• [Presentation] ニワトリ胚毛様体神経節杯状シナプス前終末における光遺伝学的操作・計測 (2013)
  • Author(s): 江川遼、細島頌子、侯旭濱、加藤秀理、石塚徹、仲村春和、八尾 寛
  • Organizer: 第36回日本神経科学大会
  • Place of Presentation: 京都
• [Book] Optogenetics: Light-Sensing Proteins and Their Applications (2015)
  • Author(s): Yawo H, Kandori H, Koizumi Eds.
  • Total Pages: 512
  • Publisher: Springer, Tokyo
• [Book] “Optogenetics: Light-Sensing Proteins and Their Applications”, Chaptor 8. General description: Future prospects of optogenetics. (2015)
  • Author(s): Yawo, H., Egawa, R., Hososhima, S., Wen, L.
  • Total Pages: 22
  • Publisher: Springer, Tokyo
• [Book] “Optogenetics: Light-Sensing Proteins and Their Applications”, Chaptor 21. Strategies to probe mechanoreception: from mechanical to optogenetic approaches. (2015)
  • Author(s): Ji, Z.-G., Ishizuka, T., Yawo, H
  • Total Pages: 10
  • Publisher: Springer, Tokyo
• [Book] オプトジェネティクス―光工学と遺伝学による行動制御技術の最前線 (2013)
  • Author(s): 八尾 寛
  • Publisher: エヌ・ティー・エス
• [Book] Optogenetics (2013)
  • Author(s): Oertner TG, Helmchen F, de Lecea L, Beck H, Konnerth A, Kaupp B, Knopfel T, Yawo H, Musser M
  • Publisher: De Gruyter, Dahlem Workshop Reports, Berlin
• [Remarks] 東北大学研究者紹介
  • URL: http://db.tohoku.ac.jp/whois/detail/0684d30b13c056e926b65120463918fb.html
• [Remarks] 東北大学大学院生命科学研究科
  • URL: http://www.lifesci.tohoku.ac.jp/
• [Remarks] 脳機能解析分野
  • URL: http://neuro.med.tohoku.ac.jp/
• [Remarks] 東北大学脳科学センター
  • URL: http://www.bsc.tohoku.ac.jp/contents/c2_32/c2_32_yao_h.html
• [Remarks] 東北大学プレスリリース
  • URL: http://www.tohoku.ac.jp/japanese/2015/02/press20150205-01.html
• [Remarks] 東北大学研究者紹介
  • URL: http://db.tohoku.ac.jp/whois/detail/0684d30b13c056e926b65120463918fb.html
• [Remarks] 東北大学大学院生命科学研究科
  • URL: http://www.lifesci.tohoku.ac.jp/teacher/t_yawo/
• [Remarks] 脳機能解析分野
  • URL: http://neuro.med.tohoku.ac.jp/