経路選択的光遺伝学機能操作法による行動制御

• Project Area: Mesoscopic neurocircuitry: towards understanding of the functional and structural basis of brain information processing
• Project/Area Number: 25115731
• Research Category: Grant-in-Aid for Scientific Research on Innovative Areas (Research in a proposed research area)
• Allocation Type: Single-year Grants
• Review Section: Biological Sciences
• Research Institution: National Institute for Physiological Sciences
• Principal Investigator: 伊佐 正 生理学研究所, 発達生理学研究系, 教授 (20212805)
• Project Period (FY): 2013-04-01 – 2015-03-31
• Project Status: Discontinued (Fiscal Year 2014)
• Budget Amount: ¥11,700,000 (Direct Cost: ¥9,000,000、Indirect Cost: ¥2,700,000); Fiscal Year 2014: ¥5,850,000 (Direct Cost: ¥4,500,000、Indirect Cost: ¥1,350,000); Fiscal Year 2013: ¥5,850,000 (Direct Cost: ¥4,500,000、Indirect Cost: ¥1,350,000)
• Keywords: 上丘 / 指向運動 / 逃避行動 / ウィルスベクター / 光遺伝学 / 脳幹網様体

Outline of Annual Research Achievements

げっ歯類において、中脳上丘から反対側網様体へ投射経路は視覚刺激等に対する指向運動を制御しているのに対し、同側網様体への投射経路は逃避行動を制御しているという仮説が提唱されてきた。しかし、その実験的証拠はあくまで間接的だった。本研究では、ウィルスベクターのみを用いて経路選択的にチャネルロドプシンを発現させ、それによってそれぞれの経路選択的に特異的な行動が観察されるかどうかを観察し、上記の仮説の検証を目的としている。今回、そのための第一歩として、AAV1-CAG-ChR2(H134R)-tdTomatoをマウスの片側上丘に注入し、５－７週間後に波長473nmの青色レーザーを直径250μmないしは500μmの光ファイバーを上丘の直上に埋めて照射し、行動を観察した。すると、直径250μmで尾外側部を刺激した際には、反対側への頭部の指向運動が誘発され、２－３秒にわたる長い時間の刺激では全身の回旋運動が誘発された。それに対し、吻内側部の刺激では上方向に頭を向ける指向運動様の動きが観察された。一方で、直径500μmの光ファイバーで上丘全体を強く刺激した際には、全身の逃避行動が観察された。次に、第二段階として、左の橋・延髄網様体に逆行性のウィルスベクターであるFuGC- MSCV-Creを注入し、その後右側の上丘にAAV-EF1α-double flex hChR2-YFPを注入し、交叉性上丘―網様体投射経路に選択的にチャネルロドプシンを発現させた。数か月以降に上丘に波長473 nmの青色レーザー光を照射したところ、反対側の頭部の指向運動を誘発できた。その後麻酔下でも上丘ニューロンがレーザー刺激に発火応答していることが確認できた。このようにして、遺伝子改変動物を作製することなく、２種類のウィルスベクターの組み合わせのみで、経路選択的な光遺伝学的手法によって、行動に影響を与えることができた。

Research Progress Status

26年度が最終年度であるため、記入しない。

Strategy for Future Research Activity

26年度が最終年度であるため、記入しない。

Research Products

• [Journal Article] Decoding grasp force profile from electrocorticography signals in non-human primate sensorimotor cortex. (2014)
  • Author(s): Chen, C., Shin, D., Watanabe, H., Nakanishi, Y., Kambara, H., Yoshimura, N., Nambu, A. and Isa, T., Nishimura, Y. & Koike, Y.
  • Journal Title: Neurosci Res Volume: S0168-0102(14). Pages: 00047-9
  • DOI: 10.1016/j.neures.2014.03.010
  • Peer Reviewed / Open Access
• [Journal Article] Cervical Dystonia: a disorder of the midbrain network for covert attentional orienting. (2014)
  • Author(s): Hutchinson M, Isa T, Molloy A, Williams L, Kimmich O, Molloy F, Moore H, Healy DG, Lynch T, Walsh C, Butler J, Reilly R, Walsh R, O’Riordan S
  • Journal Title: Frontiers in Neurology Volume: なし
  • DOI: 10.3389/fneur.2014.00054
  • Peer Reviewed
• [Journal Article] Distinct local circuit properties of the superficial and intermediate layers of the rodent superior colliculus. (2014)
  • Author(s): Phongphanphanee, P., Marino, R., Kaneda, K., Yanagawa, Y., Munoz, D.P. and Isa, T.
  • Journal Title: Eur J Neurosci. Volume: 40（2） Issue: 2 Pages: 2329-43
  • DOI: 10.1111/ejn.12579
  • Peer Reviewed / Open Access
• [Journal Article] Reconstruction of intracortical whisker-evoked local field potential from electrocorticogram using a model trained forspontaneous activity in the rat barrel cortex. (2014)
  • Author(s): Watanabe H, Sakatani T, Suzuki T, Sato MA, Nishimura Y, Nambu A, Kawato M, Isa T
  • Journal Title: Neurosci Res. Volume: S0168-0102(14) Pages: 00121-7
  • Peer Reviewed
• [Journal Article] Decoding of the spike timing of primary afferents during voluntary arm movements in monkeys. (2014)
  • Author(s): Umeda T, Watanabe H, Sato MA, Kawato M, Isa T, Nishimura Y
  • Journal Title: Frontiers in Neuroscience Volume: なし Pages: 97-97
  • DOI: 10.3389/fnins.2014.00097
  • Peer Reviewed / Open Access
• [Journal Article] Reorganization of sensory pathways after neonatal hemidecortication in rats. (2014)
  • Author(s): Wanakhachornkrai, O., Umeda, T., Isa, K., Tantisira, M., Tantisira, B. and Isa, T.
  • Journal Title: Neuroscience Research Volume: 79 Pages: 94-98
  • DOI: 10.1016/j.neures.2013.11.003
  • Peer Reviewed
• [Journal Article] Plasticity for recovery after partial spinal cord injury-Hierarchical organization- (2014)
  • Author(s): Isa, T. and Nishimura, Y.
  • Journal Title: Neuroscience Research Volume: 78 Pages: 3-8
  • DOI: 10.1016/j.neures.2013.10.008
  • Peer Reviewed
• [Journal Article] Viral vector-mediated selective and reversible blockade of the pathway for visual orienting in mice. (2013)
  • Author(s): Sooksawate T, Isa K, Matsui R, Kato S, Kinoshita M, Kobayashi K, Watanabe D, Kobayashi K, Isa T.
  • Journal Title: Frontiers in Neural Circuits Volume: 7 Pages: 162-162
  • DOI: 10.3389/fncir.2013.00162
  • Peer Reviewed
• [Journal Article] Effects of early versus late rehabilitative training on manual dexterity after corticospinal tract lesion in macaque monkeys. (2013)
  • Author(s): Sugiyama Y, Higo N, Yoshino-Saito K, Murata Y, Nishimura Y, Oishi T, Isa T
  • Journal Title: Jornal of Neurophysiology Volume: 109 Issue: 12 Pages: 2853-2865
  • DOI: 10.1152/jn.00814.2012
  • Peer Reviewed
• [Journal Article] Differential Expression of Secreted Phosphoprotein l in the Motor Cortex among Primate Species and during Postnatal Development and Functional Recovery (2013)
  • Author(s): T. Yamamoto, T. Oishi, N. Higo, S. Murayama, A. Sato, I. Takashima, Y. Sugiyama, Y. Nishimura, Y. Murata, K. Yoshino-Saito, T. Isa, T. Kojima
  • Journal Title: PLOS ONE Volume: 8(5) Issue: 5 Pages: 1-13
  • DOI: 10.1371/journal.pone.0065701
  • Peer Reviewed
• [Journal Article] GABAergic mechanisms for shaping transient visual responses in the mouse superior colliculus. (2013)
  • Author(s): Kaneda K, Isa T
  • Journal Title: Neuroscience Volume: 235 Pages: 129-140
  • DOI: 10.1016/j.neuroscience.2012.12.061
  • Peer Reviewed
• [Journal Article] DNA Methylation and Methyl-Binding Proteins Control Differential Gene Expression in Distinct Cortical Areas of Macaque Monkey. (2013)
  • Author(s): Katsusuke Hata, Hiroaki Mizukami, Osamu Sadakane, Akiya Watakabe, Masanari Ohtsuka, Masafumi Takaji, Masaharu Kinoshita, Tadashi Isa, Keiya Ozawa, and Tetsuo Yamamori
  • Journal Title: The Journal of Neuroscience Volume: 33 Issue: 50 Pages: 19704-19714
  • DOI: 10.1523/jneurosci.2355-13.2013
  • Peer Reviewed
• [Journal Article] Prediction of hand trajectory from electrocorticography signals in primary motor cortex. (2013)
  • Author(s): Chen, D., Shin, D., Watanabe, H., Nakanishi, Y., Kambara, H., Yoshimura, N., Nambu, A., Isa, T., Nishimura, Y. and Koike, Y.
  • Journal Title: PLos One Volume: 8 Issue: 12 Pages: 1-10
  • DOI: 10.1371/journal.pone.0083534
  • Peer Reviewed
• [Journal Article] Roles of direct vs indirect pathways from the motor cortex to spinal motoneurons in the control of hand dexterity. (2013)
  • Author(s): Isa, T., Kinoshita, M. and Nishimura, Y.
  • Journal Title: Frontiers in Neurology Volume: 4 Pages: 191-191
  • DOI: 10.3389/fneur.2013.00191
  • Peer Reviewed
• [Presentation] マウス上丘浅層におけるニューロン集団活動の空間的解析 (2014)
  • Author(s): 笠井昌俊，伊佐正
  • Organizer: バイオイメージ・インフォマティクスワークショップ２０１４
  • Place of Presentation: 岡崎コンファレンスセンター(愛知県、岡崎市）
  • Year and Date: 2014-06-09
• [Presentation] 障害脳が示す新たな機能 (2014)
  • Author(s): 伊佐正
  • Organizer: 第55回日本神経学会学術大会
  • Place of Presentation: 福岡国際会議場(福岡県、福岡市）
  • Year and Date: 2014-05-22
  • Invited
• [Presentation] Pathway-selective blocking technique with double viral vectors for dissecting large-scaled neural network (2014)
  • Author(s): Tadashi Isa
  • Organizer: Emerging technologies for exploring the normal and epileptic brain A Segerfalk-Pufendorf Symposium
  • Place of Presentation: Lund University(Sweden)
  • Year and Date: 2014-05-06
  • Invited
• [Presentation] Estimation of Firing of Primary Afferents from Joint Kinematics During Voluntary Arm Movements of Monkeys
  • Author(s): Umeda T, Sato M, Kawato M, Isa T, Nishimura Y
  • Organizer: 第35回国際IEEE大会
  • Place of Presentation: 大阪国際会議場(大阪）
  • Invited
• [Presentation] Selective blocakde of crossed tecto-reticular pathway by double infection of viral vectors impaired orienting responses in mice; methodological considerations.
  • Author(s): Kaoru Isa, Thongchai Sooksawate, Ryusuke Matsui, Masaharu Kinoshita, Shigeki Kato, Kenta Kobayashi, Dai Watanabe, Kazuto Kobayashi, Tadashi Isa
  • Organizer: Gordon Research Conference
  • Place of Presentation: Stonehill College(MA, USA)
• [Presentation] Acquisition of novel behaviour by visual reinforcement without awareness: instrumental learning in blindsight monkeys
  • Author(s): Rikako Kato, Norihiro Takakuwa, Abdelhafid Zeghbib, Peter Redgrave, Tadashi Isa
  • Organizer: Gordon Research Conference
  • Place of Presentation: Stonehill College(MA, USA)
• [Presentation] Verified contribution of propriospinal neurons to recovery of hand dexterity after a corticospinal tract lesion in the monkey
  • Author(s): T.Tohyama, M.Kinoshita, R.Matsui, S.Kato, K.Isa, D.Watanabe, K.Kobayashi, M.Liu, T.Isa
  • Organizer: Neuroscience2013
  • Place of Presentation: サンディエゴ国際会議場（USA)
• [Presentation] Process of forming novel behavior by visual reinforcement without awareness: Instrumental conditioning in blindsight monkeys.
  • Author(s): R.Kato, N.Takakuwa, A Zeghbib, P.Redgrave, T.Isa
  • Organizer: Neuroscience2013
  • Place of Presentation: サンディエゴ国際会議場（USA）
• [Presentation] Classical conditioning reinforced by visual stimuli in blindsight monkeys.
  • Author(s): N.Takakuwa, R.Kato, P.Redgrave, T.Isa
  • Organizer: Neuroscience2013
  • Place of Presentation: サンディエゴ国際会議場（USA）
• [Presentation] Key elements for functional recovery from the partial spinal cord injury; spinal cord, cortex and beyond
  • Author(s): Tadashi Isa
  • Organizer: Weekly Colloquium, The Winifred Masterson Burke Medical Research Institute (NY,USA)
  • Place of Presentation: The Winifred Masterson Burke Medical Research Institute (NY, USA)
  • Invited
• [Presentation] Key elements for functional recovery after partial spinal cord injury; spinal cord, cortex and beyond
  • Author(s): Tadashi Isa
  • Organizer: 15th Spinal Research Network Meeting
  • Place of Presentation: London, UK
  • Invited
• [Presentation] Dissectiong the circuit for functional recovery after neuronal damage
  • Author(s): Tadashi Isa
  • Organizer: Plenary Lecture, the 10th Biennial Conference of the Chinese Neuroscience Society
  • Place of Presentation: Beijin International Convention & Exhibition Centre
  • Invited
• [Presentation] Large-scaled network reorganization during recovery from partial spinal cord injury
  • Author(s): Tadashi Isa
  • Organizer: Distinguished Lecture Program
  • Place of Presentation: Katholic University Leuven (Belgium)
  • Invited