経路選択的光遺伝学機能操作法による行動制御

2013 Fiscal Year Annual Research Report

• 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)
• 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
• Keywords: ウィルスベクター / 上丘 / 脳幹網様体 / 指向運動 / 逃避行動

Research Abstract

マウスの中脳上丘から反対側の橋・延髄網様体に至る経路を選択的に遮断すべく、橋・延髄網様体に高頻度逆行性レンチウィルスベクターにテトラサイクリン反応性配列TREに緑色蛍光タンパクFPと破傷風毒素(eTeNT)を搭載したウィルスベクター(HiRet-TRE-eGFP.eTeNT)を注入。約１週間後に反対側上丘に高感度Tet-ON配列rtTAV16を搭載したアデノ随伴ウィルスベクター(AAV2-CMV-rtTAV16)を注入した。そして６週間後にマウスにドキシサイクリンを投与したところ、上丘の反対側に向かう頭部の指向運動、visual placing response、回旋運動が阻害された。
また、このウィルスベクター感染による選択的・可逆的経路遮断技術の技術的詳細について、このマウスの指向運動系を対象として最適化を行った。

Current Status of Research Progress

2: Research has progressed on the whole more than it was originally planned.

Reason

ウィルスベクター２重感染による経路遮断をマウスでも成功させた。そして、現在、破傷風毒素の代わりにチャネルロドプシンを搭載する、またHiRet-Tetのウィルス系だけでなく、HiRetにCre-floxの系を用いた経路選択的機能操作系の開発も行っている。

Strategy for Future Research Activity

マウスでの経路選択的機能操作の技法について確立してきたので、平成２６年度は、同側の上丘ー網様体路に遮断を適用し、さらにチャネルロドプシンを適用することで経路選択的な活性化を行い、マウスで指向運動と逃避行動を別々に制御したい。

Research Products

• [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: 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
  • DOI: doi: 10.3389/fncir.2013.00162. eCollection 2013.
  • 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: Journal of Neurophysiology Volume: 109 Pages: 2853-2865
  • DOI: doi: 10.1152/jn. 00814. 2012. Epub 2013 Mar 20.
  • Peer Reviewed
• [Journal Article] Expression of secreted phosphoprotein 1 in the motor cortex among primate species and duriing postnatal development and functional recovery. (2013)
  • Author(s): Yamamoto T, Oishi T, Higo N, Murayama S, Sato A, Takashima I, Sugiyama Y, Nishimura Y, Murata Y, Yoshino-Saito K, Isa T, Kojima T
  • Journal Title: PLos One Volume: 8 Pages: e65701
  • DOI: doi: 10.1371/journal.pone.0065701. Print 2013
  • 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: doi: 10.1016/j.neuroscience.2012.12.061. Epub 2013 Jan 18.
  • Peer Reviewed
• [Journal Article] Reorganization of sensory pathways after neonatal hemidecortication in rats. (2013)
  • 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: doi: 10.1016/j.neures.2013.11.003.Epub 2013 Nov 16.
  • Peer Reviewed
• [Journal Article] DNA methylation and methyl-binding proteins control differential gene expression in distinct cortical areas of macaque monkey. (2013)
  • Author(s): Hata, K., Mizukami, H., Sadakane, O., Watakabe, A., Kinoshita, M., Isa, T., Ozawa, K. & Yamamori, T
  • Journal Title: Journal of Neuroscience Volume: 33 Pages: 19704-19714
  • DOI: 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, Koike Y
  • Journal Title: PLos One Volume: 8 Pages: e83534
  • DOI: doi: 10.1371/journal.pone.0083534. eCollection 2013.
  • 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
  • DOI: doi: 10.3389/fneur.2013.00191
  • Peer Reviewed
• [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