記憶想起のトップダウン信号 : 新開発MRI分子プローブによる起源の同定と機能の解明

• Project/Area Number: 09J01438
• Research Category: Grant-in-Aid for JSPS Fellows
• Allocation Type: Single-year Grants
• Section: 国内
• Research Field: Neuroscience in general
• Research Institution: The University of Tokyo
• Principal Investigator: 田村 啓太 東京大学, 大学院・医学系研究科, 特別研究員(DC1)
• Project Period (FY): 2009 – 2011
• Project Status: Completed (Fiscal Year 2011)
• Budget Amount: ¥2,800,000 (Direct Cost: ¥2,800,000); Fiscal Year 2011: ¥800,000 (Direct Cost: ¥800,000); Fiscal Year 2010: ¥800,000 (Direct Cost: ¥800,000); Fiscal Year 2009: ¥1,200,000 (Direct Cost: ¥1,200,000)
• Keywords: 光遺伝学 / サル / MRI / 電気生理学 / 神経回路 / 可視化 / 制御 / 神経活動 / 脳機能 / イメージング

Research Abstract

本研究では、脳の特定部位の神経活動を人為的に制御した際に他の脳部位の神経活動に現れる影響を観察することでその部位間の機能的関係を明らかにし、全脳的な機能的相互作用に基づく記憶想起のメカニズムを解明することを目指している。
2011年度は、神経活動を高速かつ可逆的に制御できる光遺伝学的手法のサルの脳への適用に取り組んだ。まず試験的にラット小脳へこの手法を適用することで血液循環機能を双方向性に高い時間分解能で光制御出来ることに成功し、動物個体レベルでの光制御のための条件を検討することができた。サル脳深部の標的部位にウイルスベクターを注入し遺伝子導入するために、小型脳定位精密注入システムを開発した。また、昨年度ラット用として開発を始めた低侵襲性オプトロードをサル脳深部用へと発展させ、より高効率な光入出力が可能な新型に改良した。これらを総合的に動員した結果、サル脳の標的部位に光遺伝学的遺伝子を導入し、蛍光測定により動物生存中に遺伝子発現をモニタリングしながら、神経活動を光制御することを実現した。さらに、MRIで生理機能を解析するための手法として細胞内部に導入可能なMRI分子プローブの開発を進め、蛍光撮影とMRI撮像によるin vivo dual modality molecular imagingにより、マウス生体内において分子プローブの時空間動態を可視化することにも成功した。
このように、本年度は、脳機能の制御および観察のための複合的手法を整え、サルを用いて記憶想起神経回路の動作原理の解明を目指す上で必要なステップを着実に進めることができた。これらは異なる研究目的にも応用可能な基幹的な研究成果であり、学会および論文にて発表した(第13項参照)。

Research Products

• [Journal Article] Optogenetic inhibition of Purkinje cell activity reveals cerebellar control of blood pressure during postural alterations in anesthetized rats (2012)
  • Author(s): Tsubota, T., Ohashi, Y., Tamura, K., Miyashita, Y.
  • Journal Title: Neuroscience Volume: (掲載確定)(印刷中)
  • Peer Reviewed
• [Journal Article] Optogenetic manipulation of cerebellar purkinje cell activity in vivo (2011)
  • Author(s): Tsubota, T., Ohashi, Y., Tamura, K., Sato, A., Miyashita, Y.
  • Journal Title: PLoS One Volume: 6
  • Peer Reviewed
• [Journal Article] Method for Enhancing Cell Penetration of Gd3+-based MRI Contrast Agents by Conjugation with Hydrophobic Fluorescent Dyes (2011)
  • Author(s): Yamane, T., Hanaoka, K., Muramatsu, Y., Tamura, K., Adachi, Y., Miyashita, Y., Hirata, Y., Nagano, T.
  • Journal Title: Bioconjugate Chemistry Volume: 22 Issue: 11 Pages: 2227-2236
  • DOI: 10.1021/bc200127t
  • Peer Reviewed
• [Journal Article] A new glass-coated tungsten optrode enclosing multiple optic fibers for fluorescence measurement, optogenetic photo-stimulation, and single-unit recording in deep brain regions (2011)
  • Author(s): Tamura, K., Ohashi, Y., Tsubota, T., Takeuchi, D., Hirabayashi, T., Yaguchi, M., Wang, N., Miyashita, Y.
  • Journal Title: Neuroscience Research Volume: 71supplement
  • Peer Reviewed
• [Journal Article] Selective visualization of rodent and macaque neural circuits using transsynaptic and stationary markers driven by a 2A-based bicistronic lentiviral vector (2011)
  • Author(s): Ohashi, Y., Tsubota, T., Sato, A., Koyano, K.W., Tamura, K., Matsuyama, M., Yaguchi, M., Wang, N., Miyashita, Y.
  • Journal Title: Neuroscience Research Volume: 71supplement Pages: e411-e411
  • DOI: 10.1016/j.neures.2011.07.1801
  • Peer Reviewed
• [Journal Article] Development of an optogenetic method for in vivo manipulation of cerebellar Purkinje cell activity (2011)
  • Author(s): Tsubota T, Ohashi Y, Tamura K, Sato A, Matsuyama M, Miyashita Y
  • Journal Title: Neuroscience Research Volume: 71supplement Pages: e311-e311
  • DOI: 10.1016/j.neures.2011.07.1357
  • Peer Reviewed
• [Journal Article] Similarities and differences of functional connectivity as measured by spontaneous correlation of fMRI signals and effective connectivity as measured by simultaneous intracortical microstimulation and fMRI (2011)
  • Author(s): Matsui, T., Koyano, K.W., Tamura, K., Watanabe, T., Takeuchi, D., Adachi, Y., Osada, T., Miyashita, Y.
  • Journal Title: Neuroscience Research Volume: 71supplement Pages: e88-e88
  • DOI: 10.1016/j.neures.2011.07.377
  • Peer Reviewed
• [Journal Article] Sensory and mnemonic demands flexibly recruit interlaminar microcircuits in macaque temporal cortex (2011)
  • Author(s): Takeuchi, D., Hirabayashi, T., Tamura, K., Miyashita, Y.
  • Journal Title: Neuroscience Research Volume: 71supplement Pages: e70-e70
  • DOI: 10.1016/j.neures.2011.07.300
  • Peer Reviewed
• [Journal Article] Reversal of Interlaminar Signal Between Sensory and Memory Processing in Monkey Temporal Cortex (2011)
  • Author(s): Takeuchi D, Hirabayashi T, Tamura K, Miyashita Y
  • Journal Title: Science Volume: 331 Pages: 1443-1447
  • Peer Reviewed
• [Journal Article] Direct Comparison of Spontaneous Functional Connectivity and Effective Connectivity Measured by Intracortical Microstimulation: An fMRI Study in Macaque Monkeys (2011)
  • Author(s): Matsui T, Tamura K, Koyano KW, Takeuchi D, Adachi Y, Osada T, Miyashita Y
  • Journal Title: Cerebal Cortex Volume: (掲載確定(印刷中))
  • Peer Reviewed
• [Journal Article] A bicistronic lentiviral vector-based method for differential transsynaptic tracing of neural circuits (2011)
  • Author(s): Ohashi Y, Tsubota T, Sato A, Koyano KW, Tamura K, Miyashita Y
  • Journal Title: Molecular and Cellular Neuroscience Volume: 46 Pages: 136-147
  • Peer Reviewed
• [Journal Article] Genetic tracing of the rat neural circuits originating from various local brain regions using a 2A-based bicistronic lentiviral vector. (2011)
  • Author(s): Ohashi Y, Tsubota T, Sato A, Koyano KW, Tamura K, Yaguchi M, Wang N, Miyashita Y
  • Journal Title: The Journal of Physiological Sciences Volume: 61
  • Peer Reviewed
• [Journal Article] Triphasic Dynamics of Stimulus-Dependent Information Flow between Single Neurons in Macague Inferior Temporal Cortex (2010)
  • Author(s): Hirabayashi T, Takeuchi D, Tamura K, Miyashita Y
  • Journal Title: Journal of Neuroscience Volume: 30 Pages: 10407-10421
  • Peer Reviewed
• [Journal Article] Polysynaptic neuronal connectivity of SI cortex revealed by simultaneous electrical microstimulation and fMRI in macaque monkeys. (2010)
  • Author(s): Matsui T, Tamura K, Koyano KW, Takeuchi D, Watanabe T, Adachi Y, Osada T, Miyashita Y
  • Journal Title: The Journal of Physiological Sciences Volume: 60
  • Peer Reviewed
• [Journal Article] Design and synthesis of a dual-modality MRI-and fluorescence-visible muscimol derivative (2009)
  • Author(s): K.Tamura, et al.
  • Journal Title: The Journal of Physiological Sciences 59(suppl.) Pages: 426-426
  • Peer Reviewed
• [Journal Article] Transneuronal circuit tracing of the rat cerebellar pathways with a bicistronic lentiviral vector co-expressing green fluorescent protein and wheat germ agglutinin (2009)
  • Author(s): Y.Ohashi, et al.
  • Journal Title: Neuroscience 2009 Neuroscience Meeting Planner, on line
  • Peer Reviewed
• [Journal Article] Development of A Liver-specific Probe for MR and NIRF Dual Imaging (2009)
  • Author(s): T.yamane, et al.
  • Journal Title: Abstracts of the World Molecular Imaging Congress, on line Pages: 659-659
  • Peer Reviewed
• [Presentation] A new glass-coated optical and electrical probe for optogenetic investigation of monkey brain functions (2012)
  • Author(s): Tamura K, Ohashi Y, Tsubota T, Takeuchi D, Hirabayashi T, Yaguchi M, Matsuyama M, Sekine T, Miyashita Y
  • Organizer: Kyoto University and The University of Tokyo Global COE Joint Symposium
  • Place of Presentation: University of Tokyo, Tokyo, Japan
  • Year and Date: 2012-01-20
• [Presentation] A lentivirus-based optogenetic method for cell-type-specific manipulation of cerebellar Purkinje cell activity in vivo (2012)
  • Author(s): Tsubota T, Ohashi Y, Tamura K, Sato A, Yaguchi M, Matsuyama M, Sekine T, Miyashita Y
  • Organizer: Kyoto University and The University of Tokyo Global COE Joint Symposium
  • Place of Presentation: University of Tokyo, Tokyo, Japan
  • Year and Date: 2012-01-20
• [Presentation] Development of a novel glass-coated optrode for monkey optogenetics (2012)
  • Author(s): Tamura K, Ohashi Y, Tsubota T, Takeuchi D, Hirabayashi T, Yaguchi M, Matsuyama M, Sekine T, Miyashita Y
  • Organizer: The University of Tokyo Global COE, Integrative Life Sciences Based on the Study of Biosignaling Mechanisms, the 5th Retreat/Symposium
  • Place of Presentation: Yatsugatake Royal Hotel, Yamanashi, Japan
• [Presentation] Cell-type-specific optogenetic manipulation of Purkinje cell activity in vivo using lentiviral vectors (2012)
  • Author(s): Tsubota T, Ohashi Y, Tamura K, Sato A, Yaguchi M, Matsuyama M, Sekine T, Miyashita Y
  • Organizer: The University of Tokyo Global COE, Integrative Life Sciences Based on the Study of Biosignaling Mechanisms, the 5th Retreat/Symposium
  • Place of Presentation: Yatsugatake Royal Hotel, Yamanashi, Japan
• [Presentation] Reciprocal interlaminar connectivity is differentially recruited during sensory and mnemonic processing in monkey temporal cortex (2011)
  • Author(s): Takeuchi D, Hirabayashi T, Tamura K, Miyashita Y
  • Organizer: The 41^<st> annual meeting of Society for Neuroscience
  • Place of Presentation: Walter E.Washington Convention Center, Washington DC, USA
  • Year and Date: 2011-11-16
• [Presentation] Selective manipulation of cerebellar Purkinje cell activity using optogenetics in vivo (2011)
  • Author(s): Tsubota T, Ohashi Y, Tamura K, Sato A, Miyashita Y
  • Organizer: The 41^<st> annual meeting of Society for Neuroscience
  • Place of Presentation: Walter E.Washington Convention Center, Washington DC, USA
  • Year and Date: 2011-11-13
• [Presentation] Selective visualization of rodent and macaque neural circuits using transsynaptic and stationary markers driven by a 2A-based bicistronic lentiviral vector (2011)
  • Author(s): Ohashi Y, Tsubota T, Sato A, Koyano KW, Tamura K, Matsuyama M, Yaguchi M, Wang N, Miyashita Y
  • Organizer: The 34th Annual Meeting of the Japan Neuroscience Society
  • Place of Presentation: PACIFICO Yokohama, Kanagawa, Japan
  • Year and Date: 2011-09-17
• [Presentation] Similarities and differences of functional connectivity as measured by spontaneous correlation of fMRI signals and effective connectivity as measured by simultaneous intracortical microstimulation and fMRI (2011)
  • Author(s): Matsui T, Koyano K W, Tamura K, Watanabe T, Takeuchi D, Adachi Y, Osada T, Miyashita Y
  • Organizer: The 34th Annual Meeting of the Japan Neuroscience Society
  • Place of Presentation: PACIFICO Yokohama, Kanagawa Japan
  • Year and Date: 2011-09-17
• [Presentation] A new glass-coated tungsten optrode enclosing multiple optic fibers for fluorescence measurement, optogenetic photo-stimulation, and single-unit recording in deep brain regions (2011)
  • Author(s): Tamura K, Ohashi Y, Tsubota T, Takeuchi D, Hirabayashi T, Yaguchi M, Wang N, Miyashita Y
  • Organizer: The 34th Annual Meeting of the Japan Neuroscience Society
  • Place of Presentation: PACIFICO Yokohama, Kanagawa, Japan
  • Year and Date: 2011-09-16
• [Presentation] Development of an optogenetic method for in vivo manipulation of cerebellar Purkinie cell activity (2011)
  • Author(s): Tsubota T, Ohashi Y, Tamura K, Sato A, Matsuyama M, Miyashita Y
  • Organizer: The 34th Annual Meeting of the Japan Neuroscience Society
  • Place of Presentation: PACIFICO Yokohama, Kanagawa, Japan
  • Year and Date: 2011-09-16
• [Presentation] Sensory and mnemonic demands flexibly recruit interlaminar microcircuits in macaaue temporal cortex (2011)
  • Author(s): Takeuchi D, Hirabayashi T, Tamura K, Miyashita Y
  • Organizer: The 34th Annual Meeting of the Japan Neuroscience Society
  • Place of Presentation: PACIFICO Yokohama, Kanagawa, Japan
  • Year and Date: 2011-09-16
• [Presentation] Genetic tracing of the rat neural circuits originating from various local brain regions using a 2A-based bicistronic lentiviral vector. (2011)
  • Author(s): Ohashi Y, Tsubota T, Sato A, Koyano KW, Tamura K, Yaguchi M, Wang N, Miyashita Y
  • Organizer: 第88回日本生理学会
  • Place of Presentation: (東日本大震災のため誌上開催)
  • Year and Date: 2011-03-30
• [Presentation] Reversal of interlaminar singal flow between sensory and mnemonic processing in monkey temporal cortex (2011)
  • Author(s): Takeuchi D, Hirabayashi T, Tamura K, Miyashita Y
  • Organizer: The University of Tokyo Global COE, Integrative Life Sciences Based on the Study of Biosignaling Mechanisms, the 4th Retreat / Symposium
  • Place of Presentation: 山梨県北杜市 八ヶ岳ロイヤルホテル
  • Year and Date: 2011-03-05
• [Presentation] Lentivector-based transsynaptic circuit tracing : visualization of rat cortico-cortical, cortico-thalamic and cerebellar efferent pathways (2011)
  • Author(s): Ohashi Y, Tsubota T, Sato A, Koyano KW, Tamura K, Yaguchi M, Wang N, Miyashita Y
  • Organizer: The University of Tokyo Global COE, Integrative Life Sciences Based on the Study of Biosignaling Mechanisms, the 4th Retreat / Symposium
  • Place of Presentation: 山梨県北杜市 八ヶ岳ロイヤルホテル
  • Year and Date: 2011-03-05
• [Presentation] In vivo connectivity analysis of the macaque brain using fMRI (2011)
  • Author(s): Matsui T, Koyano KW, Tamura K, Watanabe T, Adachi Y, Osada T, Miyashita Y
  • Organizer: The University of Tokyo Global COE, Integrative Life Sciences Based on the Study of Biosignaling Mechanisms, the 4th Retreat / Symposium
  • Place of Presentation: 山梨県北杜市八ヶ岳ロイヤルホテル
  • Year and Date: 2011-03-05
• [Presentation] 動脈硬化巣の可視化を目指したMRIプローブの開発と応用 (2011)
  • Author(s): 花岡健二郎, 山根健浩, 村松泰明, 田村啓太, 足立雄哉, 宮下保司, 平田恭信, 長野哲雄
  • Organizer: 日本ケミカルバイオロジー学会第6回年会
  • Place of Presentation: 東京工業大学、東京、日本
• [Presentation] Comparison of spontaneous functional connectivity and effective connectivity as measured by electrical microstimulation in macaque monkeys (2010)
  • Author(s): Matsui T, Tamura K, Koyano KW, Takeuchi D, Watanabe T, Adachi Y, Osada T, Miyashita Y
  • Organizer: 米国神経科学大会2010
  • Place of Presentation: San Diego Convention Center, San Diego, USA
  • Year and Date: 2010-11-16
• [Presentation] Information flow between single neurons in macaque inferior temporal cortex : Granger causality analysis for spike trains (2010)
  • Author(s): HIRABAYASHI T, TAKEUCHI D, TAMURA K, MIYASHITA Y
  • Organizer: 米国神経科学大会2010
  • Place of Presentation: San Diego Convention Center, San Diego, USA
  • Year and Date: 2010-11-13
• [Presentation] Development of Novel Cell-permeable Gd3+-based MRI Contrast Agents (2010)
  • Author(s): Yamane T, Hanaoka K, Tamura K, Adachi Y, Miyashita Y, Nagano T
  • Organizer: EMBO Conference Series-Chemical Biology 2010
  • Place of Presentation: European Molecular Biology Laboratory, Heiderberg, Germany
  • Year and Date: 2010-09-24
• [Presentation] 動脈硬化診断を目指したMRIプローブの開発 (2010)
  • Author(s): 花岡健二郎, 村松泰明, 山根健浩, 田村啓太, 足立雄哉, 宮下保司, 平田恭信, 長野哲雄
  • Organizer: 第5回日本分子イメージング学会総会・学術集会
  • Place of Presentation: 滋賀県大津市 滋賀県立県民交流センター
  • Year and Date: 2010-05-23
• [Presentation] 新規細胞膜透過性ガドリニウム系MRI造影剤の開発と応用 (2010)
  • Author(s): 山根健浩, 花岡健二郎, 村松泰明, 田村啓太, 足立雄哉, 宮下保司, 長野哲雄
  • Organizer: 第5回日本分子イメージング学会総会・学術集会
  • Place of Presentation: 滋賀県大津市 滋賀県立県民交流センター
  • Year and Date: 2010-05-23
• [Presentation] Polysynaptic neuronal connectivity of S1 cortex revealed by simultaneous electrical microstimulation and fMRI in macaque monkeys (2010)
  • Author(s): Matsui T, Tamura K, Koyano KW, Takeuchi D, Watanabe T, Adachi Y, Osada T, Miyashita Y
  • Organizer: 第87回日本生理学会
  • Place of Presentation: 岩手県盛岡市 盛岡市民文化ホール
  • Year and Date: 2010-05-20
• [Presentation] ガドリニウム系MRI造影剤の新規細胞内導入法の開発 (2010)
  • Author(s): 山根健浩, 花岡健二郎, 村松泰明, 田村啓太, 足立雄哉, 宮下保司, 長野哲雄
  • Organizer: 日本ケミカルバイオロジー学会第5回年会
  • Place of Presentation: 神奈川県横浜市 慶應義塾大学
  • Year and Date: 2010-05-19
• [Presentation] 同時微小電気刺激・機能的磁気共鳴画像法による麻酔下マカクサル一次体性感覚野の機能的結合解析 (2010)
  • Author(s): 松井鉄平, 小谷野賢治, 田村啓太, 渡辺朋美, 竹内大吾, 足立雄哉, 長田貴宏, 宮下保司
  • Organizer: 第10回東京大学生命科学シンポジウム
  • Place of Presentation: 東京都文京区 東京大学
  • Year and Date: 2010-05-01
• [Presentation] 新規細胞膜透過性MRIプローブの開発 (2010)
  • Author(s): 山根健浩, et al.
  • Organizer: 日本薬学会第130年会
  • Place of Presentation: 岡山コンベンションセンター, 岡山大学, 岡山
  • Year and Date: 2010-03-29
• [Presentation] 動脈硬化診断を目指したMRイメージングプローブの開発 (2010)
  • Author(s): 村松泰明, et al.
  • Organizer: 日本薬学会第130年会
  • Place of Presentation: 岡山コンベンションセンター, 岡山大学, 岡山
  • Year and Date: 2010-03-29
• [Presentation] Effective connectivity in the somatosensory system of macaque monkeys revealed by simulatenous electrical microstimulation and functional magnetic resonance imaging (2010)
  • Author(s): T.Matsui, et al.
  • Organizer: The University of Tokyo Global COE, Integrative Life Sciences Based on the Study of Biosignaling Mechanisms, the 3rd Retreat/Symposium
  • Place of Presentation: The University of Tokyo, Tokyo Japan
  • Year and Date: 2010-03-05
• [Presentation] Development of a lentivirus-based method to trace neural circuits and its application to the analysis of the rat cerebellar pathways (2010)
  • Author(s): Y.Ohashi, et al.
  • Organizer: The University of Tokyo Global COE, Integrative Life Sciences Based on the Study of Biosignaling Mechanisms, the 3rd Retreat/Symposium
  • Place of Presentation: The University of Tokyo, Tokyo Japan
  • Year and Date: 2010-03-05
• [Presentation] Transneuronal circuit tracing of the rat cerebellar pathways with a bicistronic lentiviral vector co-expressing green fluorescent protein and wheat germ agglutinin (2009)
  • Author(s): Y.Ohashi, et al.
  • Organizer: Neuroscience 2009
  • Place of Presentation: McCormick Place, Chicago USA
  • Year and Date: 2009-10-19
• [Presentation] Development of A Liver-specific Probe for MR and NIRF Dual Imaging (2009)
  • Author(s): 山根健浩, et al.
  • Organizer: 2009 World Molecular Imaging Congress
  • Place of Presentation: Montreal Canada
  • Year and Date: 2009-09-25
• [Presentation] 新規肝特異的MRイメージングプローブの開発 (2009)
  • Author(s): 山根健浩, et al.
  • Organizer: 第24回生体機関連化学シンポジウム
  • Place of Presentation: 九州大学, 博多
  • Year and Date: 2009-09-14
• [Presentation] Design and synthesis of a dual-modality MRI- and fluorescence-visible muscimol derivative (2009)
  • Author(s): K.Tamura, et al.
  • Organizer: The 36th Congress of the International Union of Physiological Sciences
  • Place of Presentation: Kyoto International Conference Center, Kyoto Japan
  • Year and Date: 2009-07-31
• [Presentation] 蛍光色素の化学的特性を利用した新規機能性MRI-蛍光複合機能イメージングプローブの開発 (2009)
  • Author(s): 山根健浩, et al.
  • Organizer: 日本ケミカルバイオロジー学会第4回年会
  • Place of Presentation: 神戸ハーバーランド, 神戸
  • Year and Date: 2009-05-18
• [Presentation] シアニン色素の化学的特性を利用した肝特異的MRI-近赤外蛍光デュアルイメージングプローブの開発 (2009)
  • Author(s): 山根健浩, et al.
  • Organizer: 第4回日本分子イメージング学会総会・学術集会
  • Place of Presentation: 学術総合センター, 東京
  • Year and Date: 2009-05-14