感覚入力特異的なスクラップ&ビルドによる大脳皮質内の多細胞ネットワーク形成機構

• Project Area: Dynamic regulation of brain function by Scrap & Build system
• Project/Area Number: 16H06460
• 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: 吉村 由美子 生理学研究所, 基盤神経科学研究領域, 教授 (10291907)
• Co-Investigator(Kenkyū-buntansha): 山本 亘彦 大阪大学, 生命機能研究科, 教授 (00191429)
• Project Period (FY): 2016-06-30 – 2021-03-31
• Project Status: Granted (Fiscal Year 2020)
• Budget Amount: ¥157,430,000 (Direct Cost: ¥121,100,000、Indirect Cost: ¥36,330,000); Fiscal Year 2020: ¥27,950,000 (Direct Cost: ¥21,500,000、Indirect Cost: ¥6,450,000); Fiscal Year 2019: ¥28,470,000 (Direct Cost: ¥21,900,000、Indirect Cost: ¥6,570,000); Fiscal Year 2018: ¥28,080,000 (Direct Cost: ¥21,600,000、Indirect Cost: ¥6,480,000); Fiscal Year 2017: ¥29,640,000 (Direct Cost: ¥22,800,000、Indirect Cost: ¥6,840,000); Fiscal Year 2016: ¥43,290,000 (Direct Cost: ¥33,300,000、Indirect Cost: ¥9,990,000)
• Keywords: 大脳皮質視覚野 / 視覚反応選択性 / 神経回路再編 / RNAシークエンス / 大脳皮質 / 一次視覚野 / 経験依存的発達 / 方位選択性 / 神経活動依存的マーカー / 視覚野 / 神経回路 / 遺伝子発現 / 発達

Outline of Annual Research Achievements

発達期ラットの一次視覚野に、神経活動に依存して蛍光蛋白Venusを発現する遺伝子を導入し、縦縞刺激に反応したVenus陽性細胞と反応しなかったVenus陰性細胞（錐体細胞）を標識した。その2/3層から、Venus陽性細胞・非陽性細胞をパッチピペットにより採取し、RNA抽出、ｃDNA合成・増幅、RNA-seq解析を昨年度に引き続き行った。Venus陽性細胞と陰性細胞に分けて解析した結果、一方の細胞群に強く発現する遺伝子が複数個見出された。その中の一部の遺伝子については、in situ　hybridization法によりその発現を検証した。また、実験の効率を上げるため、Venus陽性・非陽性細胞をフローサイトメトリーによるサンプリングも開始した。
また、マウス一次視覚野を対象に、2/3層にある興奮性ニューロンとPV陽性の抑制性ニューロンサブタイプが形成する神経結合の発達と経験依存性についての解析を行った。視覚野のスライス標本上にある興奮性ニューロンとPV陽性抑制性ニューロンから同時ホールセル記録を行い、このペアの神経結合を調べた。その結果、生後発達過程で、PV陽性細胞から錐体細胞への抑制性神経結合が増加し、その結果として双方向性に結合するペアが増加した。さらに、この期間に、双方向性結合を形成するペアにおいては、興奮性、抑制性どちらかの結合しかみられないペアに比べて、神経結合が強くなることを見出した。この神経結合の強化は、生後直後からの暗室飼育により抑制された。従って、興奮性・抑制性結合の結合強度の調節には生後の視覚経験が関与すると考えられる。

Current Status of Research Progress

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

Reason

本年度は縦縞の視覚刺激に反応した錐体細胞を蛍光蛋白により標識し、蛍光蛋白陽性・非陽性細胞をサンプリングして、RNAシークエンスを行うことで、縦縞方向に反応したニューロン特異的な遺伝子を特定した。
また、一次視覚野の興奮性・抑制性結合強度は生後発達期に経験依存的に調整されることを見出した。
以上の成果により、計画はおおむね順調に進展していると判断した。

Strategy for Future Research Activity

今後は視覚刺激特異的に発現が認められた遺伝子をさらに特徴づける。また、興奮性細胞と抑制性細胞の神経結合形成を調節する分子メカニズムについての解析を推進する。さらに様々な発達段階にあるin vivoマウスを用いて、一次視覚野の神経活動を記録することで、in vitro標本により特徴づけた神経回路発達と、視覚反応の発達を対応付ける予定である。

Research Products

• [Journal Article] miR-124 dosage regulates prefrontal cortex function by dopaminergic modulation. (2019)
  • Author(s): Kozuka T, Omori Y, Watanabe S, Tarusawa E, Yamamoto H, Chaya T, Furuhashi M, Morita M, Sato T, Hirose S, Ohkawa Y, Yoshimura Y, Hikida T, Furukawa T.
  • Journal Title: Scientific reports Volume: Mar 5;9(1) Pages: 3445-3445
  • DOI: 10.1038/s41598-019-38910-2
  • NAID: 120006579467
  • Peer Reviewed / Open Access
• [Journal Article] Experience-Dependent Development of Feature-Selective Synchronization in the Primary Visual Cortex. (2018)
  • Author(s): Ishikawa Ayako Wendy、Komatsu Yukio、Yoshimura Yumiko
  • Journal Title: J Neurosci. Volume: 38 Pages: 7852-7869
  • DOI: 10.1523/jneurosci.0027-18.2018
  • Peer Reviewed
• [Journal Article] Length of myelin internodes of individual oligodendrocytes is controlled by microenvironment influenced by normal and input-deprived axonal activities in sensory deprived mouse models (2018)
  • Author(s): Osanai Yasuyuki、Shimizu Takeshi、Mori Takuma、Hatanaka Nobuhiko、Kimori Yoshitaka、Kobayashi Kenta、Koyama Shinsuke、Yoshimura Yumiko、Nambu Atsushi、Ikenaka Kazuhiro
  • Journal Title: Glia. Volume: 66 Pages: 2514-2525
  • DOI: 10.1002/glia.23502
  • Peer Reviewed / Open Access
• [Journal Article] Higher primate-like direct corticomotoneuronal connections are transiently formed in a juvenile subprimate mammal. (2018)
  • Author(s): Murabe Naoyuki、Mori Takuma、Fukuda Satoshi、Isoo Noriko、Ohno Takae、Mizukami Hiroaki、Ozawa Keiya、Yoshimura Yumiko、Sakurai Masaki
  • Journal Title: Sci Rep. Volume: - Pages: 16536-16536
  • DOI: 10.1038/s41598-018-34961-z
  • Peer Reviewed / Open Access
• [Journal Article] Visualization of Thalamocortical Axon Branching and Synapse Formation in Organotypic Cocultures. (2018)
  • Author(s): Matsumoto N, Yamamoto N
  • Journal Title: Journal of Visualized Experiments Volume: 133
  • DOI: 10.3791/56553
  • Peer Reviewed
• [Journal Article] 層構造の形成．ニューロジェネテウィクス新時代－次世代シークエンサーが拓く新しい世界． (2018)
  • Author(s): 山本亘彦
  • Journal Title: CLINICAL NEUROSCIENCE Volume: 36 Pages: 181-183
• [Journal Article] 視床大脳皮質投射系における軸索分岐のリモデリング機構. (2018)
  • Author(s): 山本亘彦
  • Journal Title: 実験医学「脳神経回路と高次脳機能－スクラップ＆ビルドによる心の発達と脳疾患の謎を解く」榎本和生，岡部繁男編. Volume: - Pages: 46-52
• [Journal Article] Visual experience regulates the development of long-term synaptic modifications induced by low-frequency stimulation in mouse visual cortex (2017)
  • Author(s): Taketoshi Sugimura, Mariko Yamamoto, Kazumasa Yamada, Yukio Komatsu, and Yumiko Yoshimura
  • Journal Title: Neurosci Res Volume: 印刷中 Pages: 36-44
  • DOI: 10.1016/j.neures.2017.02.006
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Nucleocytoplasmic Shuttling of Histone Deacetylase 9 Controls Activity-Dependent Thalamocortical Axon Branching (2017)
  • Author(s): Ricardo Alchini, Haruka Sato, Naoyuki Matsumoto, Tomomi Shimogori, Noriyuki Sugo, Nobuhiko Yamamoto
  • Journal Title: Sci Rep Volume: 7 Pages: 6024-6024
  • DOI: 10.1038/s41598-017-06243-7
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Immunocytochemistry and fluorescence imaging efficiently identify individual neurons with CRISPR/Cas9-mediated gene disruption in primary cortical cultures (2017)
  • Author(s): Hiroto Tsunematsu, Akiko Uyeda, Nobuhiko Yamamoto, Noriyuki Sugo
  • Journal Title: BMC neurosci Volume: 18 Pages: 55-55
  • DOI: 10.1186/s12868-017-0377-9
  • Peer Reviewed / Open Access
• [Journal Article] Genome Stability by DNA polymerase β in Neural Progenitors Contributes to Neuronal Differentiation in Cortical Development (2017)
  • Author(s): Kohei Onishi, Akiko Uyeda, Mistuhiro Shida, Teruyoshi Hirayama, Takeshi Yagi, Nobuhiko Yamamoto, Noriyuki Sugo
  • Journal Title: J Neurosci. Volume: 37 Pages: 8444-8458
  • DOI: 10.1523/jneurosci.0665-17.2017
  • Peer Reviewed
• [Journal Article] Activity-dependent dynamics of the transcription factor CREB in cortical neurons revealed by single-molecule imaging (2017)
  • Author(s): Kitagawa H, Sugo N, Morimatsu M, Arai Y, Yanagida T, Yamamoto N
  • Journal Title: J Neurosci Volume: 37 Pages: 1-10
  • DOI: 10.1523/jneurosci.0943-16.2016
  • Peer Reviewed
• [Journal Article] Establishment of high reciprocal connectivity between clonal cortical neurons is regulated by the Dnmt3b DNA methyltransferase and clustered protocadherins (2016)
  • Author(s): Tarusawa E., Sanbo M., Okayama A., Miyashita T., Kitsukawa T., Hirayama T., Hirabayashi T., Hasegawa S., Kaneko R., Toyoda S., Kobayashi T., Kato-Itoh M., Nakauchi H., Hirabayashi M., Yagi T., Yoshimura Y.
  • Journal Title: BMC Biol Volume: 14 Pages: 103-103
  • DOI: 10.1186/s12915-016-0326-6
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Visualization of HDAC9 Spatiotemporal Subcellular Localization in Primary Neuron Cultures (2016)
  • Author(s): Sugo N, Yamamoto N
  • Journal Title: Methods Mol Biol Volume: 1436 Pages: 119-127
  • DOI: 10.1007/978-1-4939-3667-0_9
  • Peer Reviewed / Acknowledgement Compliant
• [Presentation] The roles of visual experience in the maturation of neural responses in the visual cortex. (2018)
  • Author(s): Yumiko Yoshimura
  • Organizer: Cold Spring Harbor Asia “Latest Advances in Development & Function of Neuronal Circuits”
  • Int'l Joint Research / Invited
• [Presentation] The roles of visual experience in the maturation of secondary visual cortex. (2018)
  • Author(s): Yumiko Yoshimura
  • Organizer: The 95th Annual Meeting of the Physiological Society Meeting
  • Invited
• [Presentation] Maintenance of genome stability by DNA polymerase β is necessary for postnatal development in cortical and hippocampal neurons (2018)
  • Author(s): Uyeda, A., Onishi, K., Hirayama, T., Yagi, T., Yamamoto, N., Sugo, N.
  • Organizer: The 41st Annual Meeting of the Japan Neuroscience Society
• [Presentation] BDNF promoter activity is modulated by patterned firing activity in upper layer neurons of the developing cortex (2018)
  • Author(s): Miyasaka, Y., Yamamoto, N
  • Organizer: The 41st Annual Meeting of the Japan Neuroscience Society
• [Presentation] Glial cell dynamics associated with ectopic orticofugal projections after hemispherectomy (2018)
  • Author(s): Chang, L., Yamamoto, N.
  • Organizer: The 41st Annual Meeting of the Japan Neuroscience Society
• [Presentation] 神経活動依存的な視床皮質投射の形成機構 (2018)
  • Author(s): 山本亘彦、神経活動依存的な視床皮質投射の形成機構、東京大学大学院医学系研究科セミナー
  • Organizer: 東京大学大学院医学系研究科セミナー
  • Invited
• [Presentation] Molecular mechanisms of activity-dependent axon branching in neocortical circuits (2018)
  • Author(s): Yamamoto, N.
  • Organizer: The 5th International Symposium on Synaptic Transmission and Plasticity
  • Int'l Joint Research / Invited
• [Presentation] Patterned neuronal firing regulates promoter activity of brain-derived neurotrophic factor (BDNF) in individual cortical neurons (2018)
  • Author(s): Yamamoto, N.
  • Organizer: Spontaneous Activity in Brain Development
  • Int'l Joint Research / Invited
• [Presentation] Mechanisms of activity-dependent remodeling in neocortical circuits (2018)
  • Author(s): Yamamoto, N.
  • Organizer: Invited seminar in Korea Brain Science Institute
  • Int'l Joint Research / Invited
• [Presentation] 二次視覚野の発達における視覚経験の役割 (2018)
  • Author(s): 吉村由美子
  • Organizer: 第９５回日本生理学会大会
  • Invited
• [Presentation] The roles of visual experience in the maturation of neural circuits and functions in the visual cortex (2017)
  • Author(s): Yoshimura Y
  • Organizer: McGill University-NIPS Symposium
  • Invited
• [Presentation] Novel dependence of neural responses on the contrast of visual stimuli used for an orientation discrimination task in rat primary visual cortex (2017)
  • Author(s): Kimura R, Yoshimura Y
  • Organizer: The 40th Annual Meeting of the Japan Neuroscience Society
• [Presentation] Activity-Dependent Mechanisms for Thalamocortical Circuit Formation (2017)
  • Author(s): Yamamoto, N
  • Organizer: The 48th NIPS International Symposium
  • Invited
• [Presentation] 神経活動依存的な視床皮質軸索の分岐制御機構 (2017)
  • Author(s): 山本 亘彦
  • Organizer: 第39回神経組織培養研究会
  • Invited
• [Presentation] The role of neuronal activity and co-factors in CREB dynamics in cortical neurons: A single-molecule imaging study (2017)
  • Author(s): Kitagawa, H., Sugo, N. & Yamamoto, N.
  • Organizer: Cortical development Conference
  • Int'l Joint Research
• [Presentation] High reciprocal connectivity between clonal cortical neurons is established under the guidance of epigenetic regulation (2016)
  • Author(s): Tarusawa E, Sanbo M, Okayama A, Miyashita T, Kitsukawa T, Hirayama T, Hirabayashi T, Hasegawa S, Hirabayashi M, Yagi T, Yoshimura Y
  • Organizer: Society of Neuroscience 2016
  • Int'l Joint Research
• [Presentation] Experience-dependent maturation of neural circuits and functions in the secondary visual cortex of rats (2016)
  • Author(s): Yoshimura Y
  • Organizer: The 39th Annual Meeting of the Japan Neuroscience Society
  • Invited
• [Presentation] Experience-dependent development of fine-scale networks and feature-selective synchronization in the visual cortex (2016)
  • Author(s): Yoshimura Y
  • Organizer: International Symposium 2016 ‘Circuit Construction in the Mammalian Brain’
  • Int'l Joint Research / Invited
• [Presentation] Visual experience-dependent and independent development of visually-evoked synchronized firing in rat visual cortex (2016)
  • Author(s): Ishikawa AW, Komatsu Y, Yoshimura Y
  • Organizer: The 39th Annual Meeting of the Japan Neuroscience Society
• [Presentation] Synaptic connections are preferentially formed between cortical pyramidal neurons with similar orientation preference in both layers 2/3 and 5. (2016)
  • Author(s): Karaki T, Kim R, Bito H, Yoshimura Y
  • Organizer: The 39th Annual Meeting of the Japan Neuroscience Society
• [Presentation] Positive and negative regulation of activity-dependent axon branching in the developing cortex (2016)
  • Author(s): Yamamoto N
  • Organizer: The 39th Annual Meeting of the Japan Neuroscience society
  • Invited
• [Presentation] Positive and negative regulators for activity-dependent thalamocortical axon branching (2016)
  • Author(s): Yamamoto N
  • Organizer: International Symposium 2016 ‘Circuit Construction in the Mammalian Brain’
  • Int'l Joint Research / Invited
• [Presentation] Positive and negative mechanisms of activity-dependent circuit formation in the developing cortex (2016)
  • Author(s): Yamamoto N
  • Organizer: Molecular Neurobiology Program Seminar in New York University
  • Int'l Joint Research / Invited
• [Presentation] DNA polymerase β function in neural progenitors is required for postmitotic neuronal survival and differentiation in the developing cortex (2016)
  • Author(s): Ohnishi, K., Sugo, N., Toyoda, S., Hirayama, T., Yagi, T., Yamamoto, N.
  • Organizer: The 39th Annual Meeting of the Japan Neuroscience Society