Targeted gene expression in single neurons by opto-poration in vivo

2012 Fiscal Year Final Research Report

• Project/Area Number: 23650204
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Neurophysiology and muscle physiology
• Research Institution: The University of Tokyo
• Principal Investigator: KANO Masanobu 東京大学, 大学院・医学系研究科, 教授 (40185963)
• Project Period (FY): 2011 – 2012
• Keywords: 神経科学 / 2 光子イメージング / 遺伝子発現

Research Abstract

We develop a novel method to introduce exogenous genes in individual neurons of interest in the intact brain. By using a mode-locked Ti:Sapphire laser, the neuronal membrane can be perforated to deliver small molecules including fluorescent dye and DNA plasmid. We have confirmed that fluorescent dye molecules could be reliably introduced into individual neurons in the intact mouse brains. After optimization of the technique, it is expected to be able to deliver DNA plasmid and targeted exogenous gene expression in vivo.

Research Products

• [Journal Article] The endocannabinoid2-arachidonoylglycerol negatively regulates habituation by suppressing excitatory recurrent network activity and reducing long-term potentiation in the dentate gyrus (2013)
  • Author(s): Sugaya, Y., Cagniard, B., Yamazaki, M., Sakimura, K. & Kano, M.
  • Journal Title: J. Neurosci Volume: 33 Pages: 3588-3601
  • Peer Reviewed
• [Journal Article] Spatiotemporal Dynamics of Functional Clusters of Neurons in the Mouse Motor Cortex during a Voluntary Movement (2013)
  • Author(s): Hira, R., Ohkubo, F., Ozawa, K., Isomura, Y., Kitamura, K., Kano, M., Kasai, H. & Matsuzaki, M.
  • Journal Title: J. Neurosci Volume: 33 Pages: 1377-1390
  • Peer Reviewed
• [Journal Article] Dendritic calciumsignaling in cerebellar Purkinje cell (2012)
  • Author(s): Kitamura, K. & Kano, M.
  • Journal Title: Neural Networks published online
  • URL: http://www.sciencedirect.com/science/article/pii/S0893608012002110
  • Peer Reviewed
• [Journal Article] GABAergic inhibition regulates developmental synapse elimination in the cerebellum (2012)
  • Author(s): Nakayama, H., Miyazaki, T., Kitamura, K., Hashimoto, K., Yanagawa, Y.,Obata, K., Sakimura, K., Watanabe, M.& Kano, M.
  • Journal Title: Neuron Volume: 74 Pages: 384-396
  • Peer Reviewed
• [Journal Article] Organotypic coculture preparation for the study of developmental synapseelimination in mammalian brain (2012)
  • Author(s): Uesaka, N., Mikuni, T., Hashimoto, K., Hirai, H., Sakimura, K. & Kano, M.
  • Journal Title: J. Neurosci Volume: 32 Pages: 11688-11699
  • Peer Reviewed
• [Journal Article] Synapsetype-independent degradation of the endocannabinoid2-arachidonoylglycerol after retrograde synaptic suppression (2012)
  • Author(s): Tanimura, A., Uchigashima, M., Yamazaki, M., Uesaka, N., Mikuni, T., Abe, M., Hashimoto, K., Watanabe, M., Sakimura, K. & Kano, M.
  • Journal Title: Proc. Natl. Acad. Sci. USA Volume: 109 Pages: 12195-12200
  • Peer Reviewed
• [Journal Article] Synaptogenesis and synaptic elimination (2012)
  • Author(s): Kano, M. & Watanabe, M.
  • Journal Title: Handbook of Cerebellum and Cerebellar Disorders Volume: (in press)
• [Journal Article] Locally synchronized synaptic inputs (2012)
  • Author(s): Takahashi, N., Kitamura, K., Matsuo, N., Mayford, M., Kano, M., Matsuki, N.& Ikegaya, Y.
  • Journal Title: Science Volume: 335 Pages: 353-356
  • Peer Reviewed
• [Journal Article] 小脳プルキンエ細胞樹状突起活動のin vivoイメージング (2012)
  • Author(s): 喜多村和郎,橋本浩一,狩野方伸
  • Journal Title: 生体の科学 Volume: 63 Pages: 26-33
• [Journal Article] CaV2.1 in cerebellar Purkinje cellsregulates competitive excitatory synaptic wiring, cell survival, and cerebellar biochemical compartmentalization (2012)
  • Author(s): Miyazaki, T., Yamasaki, M., Hashimoto, K., Yamazaki, M., Abe, M., Usui, H., Kano, M., Sakimura, K. & Watanabe,M.
  • Journal Title: J. Neurosci Volume: 32 Pages: 1311-1328
  • Peer Reviewed
• [Journal Article] Climbing fiber synapse elimination in cerebellar Purkinje cells (2011)
  • Author(s): Watanabe, M. & Kano, M.
  • Journal Title: Eur. J. Neurosci Volume: 34 Pages: 1697-1710
• [Journal Article] Developmental switching ofperisomatic innervation from climbing fibers to basket cell fibers in cerebellarPurkinje cells (2011)
  • Author(s): Ichikawa, R., Yamasaki, M., Miyazaki, T., Konno, K., Hashimoto, K., Tatsumi, H., Inoue, Y., Kano, M. & Watanabe, M.
  • Journal Title: J. Neurosci Volume: 31 Pages: 16916-16927
  • Peer Reviewed
• [Journal Article] Postsynaptic P/Q-type Ca2+ channel in Purkinje cell mediates synaptic competition and elimination in developing cerebellum (2011)
  • Author(s): Hashimoto, K., Tsujita, M., Miyazaki, T., Kitamura, K., Yamazaki, M., Shin, H.-S., Watanabe, M., Sakimura, K. & Kano, M.
  • Journal Title: Proc. Natl. Acad. Sci. USA Volume: 108 Pages: 2447-2457
  • Peer Reviewed
• [Journal Article] In vivo two-photon uncaging of glutamate revealing the structure-function relationships of dendritic spines in the neocortex of adult mice (2011)
  • Author(s): Noguchi, J., Nagaoka, A., Watanabe, S., Ellis-Davies, G.C., Kitamura, K., Kano, M., Matsuzaki, M. & Kasai, H.
  • Journal Title: J.Physiol Volume: 589 Pages: 2447-2457
  • Peer Reviewed
• [Journal Article] A role for myosin Va in cerebellar plasticity and motor learning: a possible mechanism underlying neurological disorder in myosin Va disease (2011)
  • Author(s): Miyata, M., Kishimoto, Y., Tanaka, M., Hashimoto, K., Hirashima, N., Murata, Y., Kano, M. & Takagishi, Y.
  • Journal Title: J. Neurosci Volume: 31 Pages: 6067-6078
• [Journal Article] Calcium-dependent regulation of climbing fiber synapse elimination during postnatal cerebellardevelopment
  • Author(s): Kano, M., Nakayama, H., Hashimoto, K., Kitamura, K., Sakimura, K. & Watanabe, M.
  • Journal Title: J. Physiol Volume: (in press)
  • Peer Reviewed
• [Presentation] マウスバレル皮質における感覚シナプス入力の可視化 (2013)
  • Author(s): 喜多村和郎、狩野方伸
  • Organizer: 第90回日本生理学会大会
  • Place of Presentation: 東京
  • Year and Date: 20130327-29
• [Presentation] マウス小脳における登上線維応答の同期性とAldolaseC発現パターンとの関連 (2013)
  • Author(s): 堤新一郎、多田真弓、崎村建司、喜多村和郎、狩野方伸
  • Organizer: 第90回日本生理学会大会
  • Place of Presentation: 東京
  • Year and Date: 20130327-29
• [Presentation] Climbing fiber network refinementand desynchronization of Purkinje cellpopulation activity during postnatal cerebellar development (2013)
  • Author(s): Jean-Marc Good, Kazuo Kitamura, Kenji Sakimura, Masanobu Kano
  • Organizer: 第90回日本生理学会大会
  • Place of Presentation: 東京
  • Year and Date: 20130327-29
• [Presentation] 組織培養法と細胞種特異的遺伝子導入法を用いたシナプス刈り込みのメカニズム (2013)
  • Author(s): 上阪直史、三國貴康、平井宏和、狩野方伸
  • Organizer: 第90回日本生理学会大会
  • Place of Presentation: 東京
  • Year and Date: 20130327-29
• [Presentation] 内因性カンナビノイド2-アラキドノイルグリセロールは歯状回の興奮性シナプス伝達を抑制し発作を減弱する (2013)
  • Author(s): 菅谷佑樹、山崎真弥、崎村建司、狩野方伸
  • Organizer: 第90回日本生理学会大会
  • Place of Presentation: 東京
  • Year and Date: 20130327-29
• [Presentation] 小脳登上線維-プルキンエ細胞シナプスの発達において競合する登上線維間の相対的な強度の差と登上線維の絶対的な強度がそれぞれ異なる段階に寄与している (2013)
  • Author(s): 川田慎也、橋本浩一、山崎真弥、宮崎太輔、山崎美和子、三國貴康、渡辺雅彦、崎村建司、狩野方伸
  • Organizer: 第90回日本生理学会大会
  • Place of Presentation: 東京
  • Year and Date: 20130327-29
• [Presentation] Activity-dependent Refinement of Climbing Fiber to Purkinje Cell Synapses during Postnatal Cerebellar Development. International Symposium on "Sensory Systems and Neural Circuits" (2013)
  • Author(s): Kano, M.
  • Organizer: Celebrating the 22nd Anniversary of Odorant- Receptor Gene Discoveries
  • Place of Presentation: 東京
  • Year and Date: 2013-02-11
• [Presentation] Synapse type-independent degradation of 2-arachidonoylglycerol after retrograde synaptic suppression in the cerebellum (2012)
  • Author(s): A. TANIMURA, M. UCHIGASHIMA, M. YAMAZAKI, N. UESAKA, T. MIKUNI, K. HASHIMOTO, M. WATANABE, K. SAKIMURA, M. KANO.
  • Organizer: Neuroscience 2012
  • Place of Presentation: New Orleans, USA.
  • Year and Date: 20121013-17
• [Presentation] Possible involvement of theCa2+-dependent gene Arc in activity-dependent synapse elimination in the cerebellum (2012)
  • Author(s): T. MIKUNI, N. UESAKA, H. OKUNO, H. HIRAI, H. BITO, M. KANO.
  • Organizer: Neuroscience 2012
  • Place of Presentation: New Orleans, USA.
  • Year and Date: 20121013-17
• [Presentation] Visualization of an active ensemble of cortical neurons in vivo using a novel activity-dependent promoter E-SARE (2012)
  • Author(s): T. KAWASHIMA, K. KITAMURA, K. OHKI, K. SUZUKI, M. NONAKA, S. KAMIJO, S. TAKEMOTO-KIMURA, M. KANO, H. OKUNO, H. BITO.
  • Organizer: Neuroscience 2012
  • Place of Presentation: New Orleans, USA.
  • Year and Date: 20121013-17
• [Presentation] Electrical contribution of postsynaptic T-type calcium channels to parallelfiber-Purkinje cell synaptic transmission (2012)
  • Author(s): R. LY, V. UEBELE, J. RENGER, H.PROSSER, A. RANDALL, M. KANO, K. SAKIMURA, B. BARBOUR, P. ISOPE, A. FELTZ.
  • Organizer: Neuroscience 2012
  • Place of Presentation: New Orleans, USA.
  • Year and Date: 20121013-17
• [Presentation] Impairment ofreward-based adaptive choice of actions by local injection of cannabinoid CB1 receptor antagonistinto the striatum of behaving monkeys (2012)
  • Author(s): Y. UEDA, S. R. OKAZAKI, K.YAMANAKA, K. ENOMOTO, M. KANO, M. KIMURA.
  • Organizer: Neuroscience 2012
  • Place of Presentation: New Orleans, USA.
  • Year and Date: 20121013-17
• [Presentation] Postnatal climbing fiber synapse remodeling desynchronizes population activity of Purkinje cells in developing cerebellum (2012)
  • Author(s): Jean- Marc Good, Kazuo Kitamura, Kenji Sakimura, Masanobu Kano.
  • Organizer: 第35回日本神経科学大会
  • Place of Presentation: 名古屋
  • Year and Date: 20120918-21
• [Presentation] 新規活動依存的プロモーターE-SAREを用いた、大脳皮質における機能的回路の同定 (2012)
  • Author(s): 川島尚之、喜多村和郎、大木研一、鈴木敢三、野中美応、上條諭志、竹本さやか、狩野方伸、奥野浩行、尾藤晴彦
  • Organizer: 第35回日本神経科学大会
  • Place of Presentation: 名古屋
  • Year and Date: 20120918-21
• [Presentation] 発達期マウス網膜-外側膝状体シナプス除去における代謝型グルタミン酸受容体1型の役割 (2012)
  • Author(s): 鳴島円、内ヶ島基政、橋本浩一、饗場篤、渡辺雅彦、宮田麻理子、狩野方伸
  • Organizer: 第35回日本神経科学大会
  • Place of Presentation: 名古屋
  • Year and Date: 20120918-21
• [Presentation] 発達期小脳におけるシナプス除去カルシウム依存性調節 (2012)
  • Author(s): 狩野方伸
  • Organizer: 第35回日本神経科学大会
  • Place of Presentation: 名古屋
  • Year and Date: 20120918-21
• [Presentation] Heterogeneous organization of individual synaptic inputs in the mouse barrel cortex (2012)
  • Author(s): Kazuo Kitamura, Yuji Ikegaya, Masanobu Kano.
  • Organizer: FENS Forum 2012
  • Place of Presentation: Barcelona, Spain.
  • Year and Date: 20120714-18
• [Presentation] Spatial And Temporal Structure Of Cortical Microcircuit Activity For Generating Voluntary Movement (2012)
  • Author(s): Hira R., Ohkubo F., Ozawa K., Isomura Y., Kitamura K., Kano M., Kasai H. & Matsuzaki M.
  • Organizer: FENS Forum 2012
  • Place of Presentation: Barcelona, Spain.
  • Year and Date: 20120714-18
• [Presentation] Calcium-dependent regulation of climbing fiber synapse elimination during postnatal cerebellar development (2012)
  • Author(s): Kano, M.
  • Organizer: Satellite symposium of SFN 2012 meeting
  • Place of Presentation: New Orleans, USA.
  • Year and Date: 2012-10-12
• [Presentation] Retrograde suppression of synaptic transmission by the endocannabinoid 2-arachidonoylglycerol (2012)
  • Author(s): Kano, M.
  • Organizer: Cell Symposia
  • Place of Presentation: New Orleans, USA.
  • Year and Date: 2012-10-11
• [Presentation] Control of synaptic function by endogenous cannabinoid (2012)
  • Author(s): Kano, M.
  • Organizer: Plenary Lecture at the 8th FENS Forum of European Neuroscience
  • Place of Presentation: Barcelona, Spain.
  • Year and Date: 2012-07-16
• [Presentation] Refinement of Climbing Fiber to Purkinje Cell Synapses during Postnatal Cerebellar Development (2012)
  • Author(s): Kano, M.
  • Organizer: 1st International Syposium / 59th NIBB Conference
  • Place of Presentation: 岡崎
  • Year and Date: 2012-03-11
• [Presentation] Postsynaptic P/Q typevoltage-dependent Ca2+ channel is involved in refinement of cerebellar climbing fiber to Purkinje cell synapses in early postnatal development (2011)
  • Author(s): Hashimoto, K., Tsujita, M., Miyazaki,T., Kitamura, K., Yamazaki M., Shin,H-S, Watanabe M., Sakimura K., Kano, M.
  • Organizer: 第34回日本神経科学大会
  • Place of Presentation: 横浜
  • Year and Date: 20110914-17
• [Presentation] Roles of Purkinje cell activity in climbing fiber synapse elimination in an organotypic coculture preparation of the cerebellum and medullao blongata (2011)
  • Author(s): Mikuni, T., Uesaka, M., Kano, M.
  • Organizer: 第34回日本神経科学大会
  • Place of Presentation: 横浜
  • Year and Date: 20110914-17
• [Presentation] Enhanced synchrony of climbingfiber-induced Ca2+ signaling in cerebellar Purkinje cells caused by elevated synchronous activity of inferior olivary neurons in GluD2 (GluRδ2) knockout mouse (2011)
  • Author(s): Miki Hashizume, Kazuo, Kitamura, Kenji Sakimura and Masanobu Kano.
  • Organizer: 第34回日本神経科学大会
  • Place of Presentation: 横浜
  • Year and Date: 20110914-17
• [Presentation] Desynchronization of cerebellar Purkinje cell population activity during postnatal development (2011)
  • Author(s): Good, J.K., Kitamura, K., Kano, M.
  • Organizer: 第34回日本神経科学大会
  • Place of Presentation: 横浜
  • Year and Date: 20110914-17
• [Presentation] Requirement oftype 1 metabotropic glutamate receptor for experience-dependent pruning of retinogeniculate synapses (2011)
  • Author(s): Narushima, M., Uchigashima, M., Hashimoto, K., Aiba, A., Watanabe, M., Miyata, M., Kano, M.
  • Organizer: 8th IBRO World Congress
  • Place of Presentation: Florence, Itary.
  • Year and Date: 20110714-18
• [Presentation] Activity-dependent refinement of climbing fiber to Purkinje cell synapses during postnatal cerebellar development (2011)
  • Author(s): Kano, M., Hashimoto, K., Kawamura, Y., Nakayama, H., Kitamura, K.,Sakimura K.
  • Organizer: COLD SPRING HARBOR ASIA CONFERENCE: "Assembly, Plasticity, Dysfunction and Repair of NeuralCircuits"
  • Place of Presentation: 蘇州、中国
  • Year and Date: 2011-10-19
• [Presentation] Activity-dependent maturation of climbing fiber to Purkinje cell synapses during postnatal cerebellar development (2011)
  • Author(s): Kano, M.
  • Organizer: 4th International Symposium of the Society for Research on the Cerebellum
  • Place of Presentation: 東京
  • Year and Date: 2011-09-18
• [Presentation] Activity-dependent synapse elimination during postnatal development (2011)
  • Author(s): Kano, M.
  • Organizer: Gordon Research Conference on Cerebellum
  • Place of Presentation: New London, USA
  • Year and Date: 2011-08-24
• [Presentation] Activity-dependentrefinement of climbing fiber to Purkinje cell synapses duringpostnatal development (2011)
  • Author(s): Kano, M.
  • Organizer: Symposium on Organization and Function of Neuronal Circuits in Movement Control
  • Place of Presentation: Trolleholm Castle, Sweden
  • Year and Date: 2011-05-28