神経回路の選択的可視化と操作を実現するウイルスベクターシステムの開発

• Project Area: Mechanisms underlying the functional shift of brain neural circuitry for behavioral adaptation
• Project/Area Number: 15H01431
• 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: Kyoto University
• Principal Investigator: 井上 謙一 京都大学, 霊長類研究所, 助教 (90455395)
• Project Period (FY): 2015-04-01 – 2017-03-31
• Project Status: Completed (Fiscal Year 2016)
• Budget Amount: ¥5,850,000 (Direct Cost: ¥4,500,000、Indirect Cost: ¥1,350,000); Fiscal Year 2016: ¥2,860,000 (Direct Cost: ¥2,200,000、Indirect Cost: ¥660,000); Fiscal Year 2015: ¥2,990,000 (Direct Cost: ¥2,300,000、Indirect Cost: ¥690,000)
• Keywords: 神経科学 / 脳・神経 / バイオテクノロジー / ウイルスベクター / 霊長類 / オプトジェネティクス

Outline of Annual Research Achievements

本研究では、新規に開発する逆行性感染型ウイルスベクターや順行性感染型ウイルスベクターを利用して、特定の神経回路を構成するニューロン集団の選択的可視化や活動操作をおこなう新規手法を開発し、霊長類において神経回路の機能シフト研究をおこなうための基盤技術を確立するためのシステム開発研究を行っている。今年度は、まずジャンクションが最適化されたキメラエンベロープタンパク質を利用した逆行性感染型レンチウイルスベクター(FuG-E型)の霊長類神経系における感染動態を従来型(FuG-C型)と比較し、FuG-E型レンチウイルスベクターが霊長類においてFuG-C型の約２倍の逆行性感染能を有することを明らかにした論文を発表した。また、FuG-E型とHiRetベクターであるFuG-B2型の霊長類（マカク・マーモセット）およびげっし類（ラット）における感染動態や注入部位における免疫反応などの差異を線条体注入系と皮質注入系において明らかにした（論文準備中）。一方、独自に開発した狂犬病ウイルスベクターのゲノム改変によって作成した、ゲノム複製能や外来遺伝子発現量が異なる複数の感染伝播能欠損型ベクターの性状解析を行い、低細胞毒性型、長期発現型の伝播能欠損型狂犬病ウイルスベクターの開発を進めた。さらに、霊長類における化学遺伝学（DREADD法）の適用に成功し、薬剤（CNO）の血中投与により、吻内側尾状核ニューロンの活動を選択的に抑制すると、報酬課題において行動変化が生じることを確認した。

Research Progress Status

28年度が最終年度であるため、記入しない。

Strategy for Future Research Activity

28年度が最終年度であるため、記入しない。

Research Products

• [Int'l Joint Research] NIH(米国)
• [Journal Article] Visuomotor signals for reaching movements in the rostro-dorsal sector of the monkey thalamic reticular nucleus. (2017)
  • Author(s): Saga Y, Nakayama Y, Inoue K, Yamagata T, Hashimoto M, Tremblay L, Takada M, Hoshi E
  • Journal Title: Eur J Neurosci Volume: 印刷中 Issue: 9 Pages: 1186-1199
  • DOI: 10.1111/ejn.13421
  • NAID: 120006305932
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] The use of an optimized chimeric envelope glycoprotein enhances the efficiency of retrograde gene transfer of a pseudotyped lentiviral vector in the primate brain. (2017)
  • Author(s): Tanabe S, Inoue K, Tsuge H, Uezono S, Nagaya K, Fijiwara M, Kato S, Kobayashi K, Takada M
  • Journal Title: Neurosci Res Volume: 印刷中 Pages: 45-52
  • DOI: 10.1016/j.neures.2017.02.007
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Propagated but Topologically Distributed Forebrain Neurons Expressing Alpha-Synuclein in Aged Macaque. (2016)
  • Author(s): Kimura K,Inoue K, Kuroiwa Y, Tanaka F, Takada M
  • Journal Title: PLoS ONE Volume: 11 Issue: 11 Pages: e0166861-e0166861
  • DOI: 10.1371/journal.pone.0166861
  • NAID: 120005954462
  • Peer Reviewed / Open Access
• [Journal Article] PET imaging-guided chemogenetic silencing reveals a critical role of primate rostromedial caudate in reward evaluation. (2016)
  • Author(s): Nagai Y, Kikuchi E, Lerchner W, Inoue Ki, Ji B, Eldridge MA, Kaneko H, Kimura Y, Oh-Nishi A, Hori Y, Kato Y, Hirabayashi T, Fujimoto A, Kumata K, Zhang MR, Aoki I, SuharaT, Higuchi M, Takada M, Richmond BJ, Minamimoto T.
  • Journal Title: Nature Communications Volume: 7 Issue: 1 Pages: 13605-13605
  • DOI: 10.1038/ncomms13605
  • NAID: 120005898960
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Origins of multisynaptic projections from the basal ganglia to the forelimb region of the ventral premotor cortex in macaque monkeys. (2016)
  • Author(s): Ishida H, Inoue K, Takada M, Hoshi E
  • Journal Title: Eur J Neurosci Volume: 43 Issue: 2 Pages: 258-269
  • DOI: 10.1111/ejn.13127
  • NAID: 120005997650
  • Peer Reviewed
• [Journal Article] Neuronal and behavioral modulations by pathway-selective optogenetic stimulation of the primate oculomotor system. (2015)
  • Author(s): Inoue K, Takada M, Matsumoto M
  • Journal Title: Nat Commun Volume: 6 Issue: 1 Pages: 8378-8378
  • DOI: 10.1038/ncomms9378
  • Peer Reviewed / Open Access
• [Journal Article] Distributions of glutamatergic, GABAergic, and glycinergic neurons in the auditory pathways of macaque monkeys. (2015)
  • Author(s): Ito T, Inoue K, Takada M
  • Journal Title: Neuroscience Volume: 310 Pages: 128-151
  • DOI: 10.1016/j.neuroscience.2015.09.041
  • Peer Reviewed / Open Access
• [Presentation] Prevention of MPTP-induced parkinsonism by recruitment of calbindin into nigral dopamine neurons. (2017)
  • Author(s): Inoue K, Miyachi S, Nishi K, Okado H, Nagai Y, Minamimoto T, Nambu A, Takada M
  • Organizer: the 12th International Basal Ganglia Society Meeting, IBAGS 2017.
  • Place of Presentation: Merida, Mexico
  • Year and Date: 2017-03-27
  • Int'l Joint Research
• [Presentation] ウイルスベクターを利用した霊長類における神経ネットワーク操作 (2017)
  • Author(s): 井上謙一
  • Organizer: 第6回 生理研-霊長研-脳研 合同シンポジウム
  • Place of Presentation: 新潟大学脳研究所統合脳機能研究センター、新潟
  • Invited
• [Presentation] 狂犬病ウイルスベクターを用いた逆行性越シナプス的ラベル法によるマーモセット帯状皮質への入力様式の解明：大脳基底核からの入力について (2017)
  • Author(s): 上園志織、柘植仁美、田辺創思、藤原真紀、長屋七奈、長屋清美、井上謙一、高田昌彦
  • Organizer: 第112回日本解剖学会総会・全国学術集会
  • Place of Presentation: 長崎大学、長崎
• [Presentation] Manipulation of primate neural networks by means of modified viral vectors (2016)
  • Author(s): Inoue K
  • Organizer: NHP Chemogenetics Workshop
  • Place of Presentation: National Institute of Health, Bethesda, USA
  • Year and Date: 2016-12-01
  • Int'l Joint Research / Invited
• [Presentation] Neuronal and behavioral modulations by pathway-selective optogenetic stimulation of the primate oculomotor system (2016)
  • Author(s): Inoue K, Takada M, Matsumoto M
  • Organizer: International Symposium on Adaptive Circuit Shift 2016
  • Place of Presentation: 同志社大学寒梅館、京都
  • Year and Date: 2016-03-03
  • Int'l Joint Research
• [Presentation] 霊長類における光遺伝学を利用した神経回路の選択的操作 (2016)
  • Author(s): 井上謙一
  • Organizer: 新潟大学脳研究所-生理研合同シンポジウム
  • Place of Presentation: 自然科学研究機構、岡崎
  • Year and Date: 2016-03-01
• [Presentation] Primate models for elucidating the circuit pathology of nigrostriatal dopamine system (2016)
  • Author(s): Inoue K
  • Organizer: 第39回日本神経科学大会サテライトシンポジウム：大脳基底核の機能と疾患：基礎と臨床
  • Place of Presentation: パシフィコ横浜，横浜
  • Int'l Joint Research / Invited
• [Presentation] Motivation center in the ventral midbrain directly activates the descending motor pathways via the primary motor cortex. (2016)
  • Author(s): Suzuki M, Inoue K, Nakagawa H, Takada M, Isa T, Nishimura Y
  • Organizer: 第39回日本神経科学大会
  • Place of Presentation: パシフィコ横浜，横浜
• [Presentation] Origin of multisynaptic projections from the amygdala to the forelimb region of the ventral premotor cortex in macaque monkeys. (2016)
  • Author(s): Ishida M, Inoue K, Takada M, Hoshi E
  • Organizer: 第39回日本神経科学大会
  • Place of Presentation: パシフィコ横浜，横浜
• [Presentation] Investigations of spontaneously naturally emerging parkinsonism-cerebellar syndromein an aged Japanese macaque (Macaca fuscata yakui): a potential analogue of multiplesystem atrophy. (2016)
  • Author(s): K.W. McCairn, Ninomiya T, Nagai Y, Go Y, Inoue K, Kimura K, Matsumoto M, Minamimoto M, Isoda M, Takada M
  • Organizer: 第39回日本神経科学大会
  • Place of Presentation: パシフィコ横浜，横浜
• [Presentation] Comparison of efficiency of retrograde gene transfer between lentiviral vectors pseudotyped with FuG-E and FuG-B2 glycoprotein in primate brains: Cortical input system. (2016)
  • Author(s): Tsuge H, Uezono S, Tanabe S, Fujiwara M, Nagaya K, Sugawara M, Miwa M, Konoike N, Kato S, Nakamura K, Kobayashi K, Inoue K, Takada M
  • Organizer: 第39回日本神経科学大会
  • Place of Presentation: パシフィコ横浜，横浜
• [Presentation] Comparison of the efficiency of retrograde gene transfer between lentiviral vectors pseudotyped with FuG-E and FuG-B2 glycoprotein in primate brains: Striatal input system. (2016)
  • Author(s): Tanabe S, Uezono S, Tsuge H, Fujiwara M, Nagaya K, Sugawara M, Miwa M, Konoike N, Kato S, Nakamura K, Kobayashi K, Inoue K,
  • Organizer: 第39回日本神経科学大会
  • Place of Presentation: パシフィコ横浜，横浜
• [Presentation] Use of an optimized chimeric envelope glycoprotein for enhancement of the efficiency of retrograde gene transfer of a pseudotyped lentiviral vector in the primate brain. (2016)
  • Author(s): Inoue K, Tanabe S, Tsuge H, Ueno T, Nagaya K, Fujiwara M, Sugawara M, Kato S, Kobayashi K, Takada M
  • Organizer: 第39回日本神経科学大会
  • Place of Presentation: パシフィコ横浜，横浜
• [Presentation] In vivo electrophysiological evaluation of chanelrhodopsin-2-expressed dorsal root ganglion neurons in adults rats. (2016)
  • Author(s): Seki K, S Wupuer, Umeda T, Inoue K, Kudo M, Takada M
  • Organizer: 第39回日本神経科学大会
  • Place of Presentation: パシフィコ横浜，横浜
• [Presentation] Differential adeno-associated virus mediated gene transfer to dorsal root ganglion neurons with different size in common marmosets. (2016)
  • Author(s): Kudo M, S Wupuer, Inoue K, Takada M, Seki K
  • Organizer: 第39回日本神経科学大会
  • Place of Presentation: パシフィコ横浜，横浜
• [Presentation] 霊長類におけるウイルスベクターを利用した神経回路の光操作 (2016)
  • Author(s): 井上謙一
  • Organizer: ルミノジェネティクス研究会
  • Place of Presentation: KKRホテル熱海、熱海
• [Presentation] Multisynaptic projections from the basal nucleus of the amygdala to the ventral premotor cortex in macaque monkeys. (2016)
  • Author(s): Ishida H, Inoue K, Takada M, Hoshi E
  • Organizer: Neuroscience 2016
  • Place of Presentation: San Diego、USA
  • Int'l Joint Research
• [Presentation] Use of an optimized chimeric envelope glycoprotein enhances the efficiency of retrograde gene transfer of a pseudotyped lentiviral vector in the primate brain. (2016)
  • Author(s): Fujiwara M, Tanabe S, Tsuge H, Uezono S, Nagaya K, Sugawara M, Kato S, Kobayashi K, *Inoue K, Takada M
  • Organizer: Neuroscience 2016
  • Place of Presentation: San Diego、USA
  • Int'l Joint Research
• [Presentation] Studying the structure and function of primate brain by using viral vectors (2016)
  • Author(s): Inoue K
  • Organizer: The 7th International Neural Microcircuit Conference
  • Place of Presentation: 自然科学研究機構 岡崎コンファレンスセンター, 岡崎
  • Int'l Joint Research / Invited
• [Presentation] The lentiviral vector pseudotyped with FuG-E glycoprotien is more suitable, compared with FuG-B2, for retrograde gene transfer in the cortical input system of primate brains. (2016)
  • Author(s): Tsuge H, Uezono S, Tanabe S, Fujiwara M, Sugawara M, Miwa M, Konoike N, Kato S, Nakamura K, Kobayashi K, Inoue K, Takada M
  • Organizer: 第6回マーモセット研究会
  • Place of Presentation: 東京大学, 東京
• [Presentation] Differences in efficiency of retrograde gene transfer and cytotoxity between lentiviral vectors pseudotyped with FuG-E and FuG-B2 glycoprotein in primate brains. (2016)
  • Author(s): Sugawara M, *Tanabe S, Uezono S, Tsuge H, Fujiwara M, Miwa M, Konoike N, Kato S, Nakamura K, Inoue K, Takada M, Kobayashi K
  • Organizer: 第6回マーモセット研究会
  • Place of Presentation: 東京大学, 東京
• [Presentation] Manipulation of primate neural networks by means of modified viral vectors. (2015)
  • Author(s): Inoue K
  • Organizer: 5th NIPS-CIN Joint Symposium
  • Place of Presentation: 生理学研究所、岡崎
  • Year and Date: 2015-11-05
  • Int'l Joint Research / Invited
• [Presentation] Development in a novel maze-task device for macaques to explore the neural mechanisms underlying motor skill learning. (2015)
  • Author(s): Yasukochi R, Inoue K, Takada M
  • Organizer: Neuroscience 2015
  • Place of Presentation: Chicago、USA
  • Year and Date: 2015-10-21
  • Int'l Joint Research
• [Presentation] Spontaneously emerging Parkinsonism-cerebellar syndrome in a subspecies of japanese macaque (macaca fuscata yakui): a potential analogue of multiple system atrophy. (2015)
  • Author(s): K.W.McCairn, Nagai Y, Kimura K, Go Y, Inoue K, Isoda M, Minamimoto T, Matsumoto M, Ninomiya T, Takada M
  • Organizer: Neuroscience 2015
  • Place of Presentation: Chicago、USA
  • Year and Date: 2015-10-19
  • Int'l Joint Research
• [Presentation] Optogenetic stimulation of the pathway from the frontal eye field to the superior colliculus evokes neuronal and behavioral modulations in monkeys. (2015)
  • Author(s): Inoue K, Takada M, Matsumoto M
  • Organizer: Neuroscience 2015
  • Place of Presentation: Chicago、USA
  • Year and Date: 2015-10-19
  • Int'l Joint Research
• [Presentation] Recruitment of calbindin into nigral dopaminergic neurons prevents from MPTP-induced parkinsonism. (2015)
  • Author(s): Takada M, Inoue K, Miyachi S
  • Organizer: 2nd World Congress on NeuroTherapeutics
  • Place of Presentation: Prague, Czech Republic
  • Year and Date: 2015-09-03
  • Int'l Joint Research
• [Presentation] Origins of multisynaptic projections from the basal ganglia to the ventral premotor cortex in macaque monkeys. (2015)
  • Author(s): Ishida H, Inoue K, Takada M, Hoshi E
  • Organizer: 第38回日本神経科学大会
  • Place of Presentation: 神戸国際会議場、神戸
  • Year and Date: 2015-07-30
• [Presentation] Development of a novel device for elucidating the neural mechanisms of motor skill learning in macaques. (2015)
  • Author(s): Yasukochi R, Inoue K, Takada M
  • Organizer: 第38回日本神経科学大会
  • Place of Presentation: 神戸国際会議場、神戸
  • Year and Date: 2015-07-29
• [Presentation] Oculomotor manipulations by pathway-selective optogenetics in nonhuman primates. (2015)
  • Author(s): Inoue K, Fujiwara M, Yasukochi R, Nagaya K, Takada M
  • Organizer: 第38回日本神経科学大会
  • Place of Presentation: 神戸国際会議場、神戸
  • Year and Date: 2015-07-29
• [Book] Gene Therapy for Neurological Disorders: Methods and Protocols, Methods in Molecular Biology, vol 1382 (Manfredsson FP, ed) (2016)
  • Author(s): Kobayashi K, Kato S, Inoue K, Takada M, Kobayashi K
  • Total Pages: 493
  • Publisher: Springer
• [Remarks] 京都大学霊長類研究所 統合脳システム分野 ホームページ
  • URL: http://www.pri.kyoto-u.ac.jp/sections/systems_neuroscience/index.html