Single Molecule Imaging of Transcription Dynamics in Neuronal Differentiation

• Project/Area Number: 17K07109
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Neurochemistry/Neuropharmacology
• Research Institution: Osaka University
• Principal Investigator: Sugo Noriyuki 大阪大学, 生命機能研究科, 特任准教授(常勤) (20372625)
• Project Period (FY): 2017-04-01 – 2022-03-31
• Project Status: Completed (Fiscal Year 2021)
• Budget Amount: ¥4,940,000 (Direct Cost: ¥3,800,000、Indirect Cost: ¥1,140,000); Fiscal Year 2019: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2018: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2017: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
• Keywords: 分子・細胞・神経生物学 / 分子・脳・神経生物学 / 1分子イメージング

Outline of Final Research Achievements

Gene expression regulation is essentially involved in the process of neuronal differentiation. However, the spatiotemporal dynamics of its regulatory factors in the nucleus remains unclear. In this study, we found the role of spatiotemporal dynamics of nuclear molecules such as transcription factors and epigenetic regulatory factors during neuronal differentiation using imaging tools in genetically modified mice and human pluripotent stem cells.

Academic Significance and Societal Importance of the Research Achievements

胎児期から生後発達期の脳機能構築において、その中心にある神経細胞の成熟には適切な遺伝子の発現調節が重要です。その過程は、核内のゲノムDNAや酵素タンパク質といった分子の相互作用によって引き起こされると考えられますが、それら分子がいつ・どこで・どのように振る舞うのかに関しては不明な点が多く残されています。本研究では、そのよう分子である転写因子とエピジェネティクス制御因子に着目してマウスおよびヒト神経細胞の成熟過程での詳細な動態や新たな役割を明らかにすることができました。この成果は、脳形成の学問的理解に貢献するとともに脳発達障害や精神神経疾患といった病態の理解に繋がることが期待されます。

Research Products

• [Journal Article] Suppression of DNA Double-Strand Break Formation by DNA Polymerase β in Active DNA Demethylation Is Required for Development of Hippocampal Pyramidal Neurons (2020)
  • Author(s): Uyeda A, Onishi K, Hirayama T, Hattori S, Miyakawa T, Yagi T, Yamamoto N, Sugo N.
  • Journal Title: J Neurosci. Volume: 40 Issue: 47 Pages: 9012-9027
  • DOI: 10.1523/jneurosci.0319-20.2020
  • Peer Reviewed
• [Journal Article] Suppression of DSB Formation by Polβ in Active DNA Demethylation is Required for Postnatal Hippocampal Development (2019)
  • Author(s): Akiko Uyeda, Kohei Onishi, Teruyoshi Hirayama, Satoko Hattori, Tsuyoshi Miyakawa, Takeshi Yagi, Nobuhiko Yamamoto, Noriyuki Sugo
  • Journal Title: bioRxiv Volume: -
  • DOI: 10.1101/852053
  • Open Access
• [Journal Article] Genome Stability by DNA polymerase β in Neural Progenitors Contributes Neuronal Differentiation in Cortical Development (2017)
  • Author(s): Kohei Onishi, Akiko Uyeda, Mistuhiro Shida, Teruyoshi Hirayama, Takeshi Yagi, Nobuhiko Yamamoto, Noriyuki Sugo
  • Journal Title: Journal of Neuroscience Volume: 37 Issue: 35 Pages: 8444-8485
  • DOI: 10.1523/jneurosci.0665-17.2017
  • Peer Reviewed
• [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 Neuroscience Volume: 18 Issue: 1 Pages: 55-55
  • DOI: 10.1186/s12868-017-0377-9
  • Peer Reviewed / Open Access
• [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: Scientific Reports Volume: 7 Issue: 1 Pages: 6024-6024
  • DOI: 10.1038/s41598-017-06243-7
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Presentation] ヒト由来大脳皮質神経細胞において共因子を伴ったCREBの動態変化が神経活動依存的に転写を促進する (2022)
  • Author(s): 渥美友梨，菅生紀之，岩田亮平，Pierre Vanderhaeghen，山本亘彦
  • Organizer: 第15回神経発生討論会
• [Presentation] ヒト由来大脳皮質神経細胞における転写調節因子CREBの共因子CBPを介した神経活動依存的な動態変化 (Activity-dependent dynamics of the transcription factor CREB via cofactor CBP in human cortical-like neurons) (2021)
  • Author(s): 渥美友梨，菅生紀之，岩田亮平，Pierre Vanderhaeghen，山本亘彦
  • Organizer: 第44回日本神経科学大会
• [Presentation] ヒト由来大脳皮質神経細胞における転写調節因子CREBと共因子CBPの神経活動依存的な動態 (Activity-dependent dynamics of the transcription factor CREB via cofactor CBP in human cortical-like neurons) (2021)
  • Author(s): 渥美友梨，菅生紀之，岩田亮平，Pierre Vanderhaeghen，山本亘彦
  • Organizer: 第43回神経組織培養研究会
• [Presentation] ヒトiPS細胞由来大脳オルガノイドにおいてDNAポリメラーゼβ欠損は神経前駆細胞にDNA2本鎖切断を引き起こす (2021)
  • Author(s): 千葉清歌、増田光起、恒松大翔、黒沢綾、足立典隆、山本亘彦、八木健、菅生紀之
  • Organizer: 第44回日本神経科学大会
• [Presentation] 体細胞核由来クローンES細胞の全ゲノム解析によるDNAポリメラーゼβ欠損マウス大脳皮質神経細胞の突然変異解析 (2021)
  • Author(s): 松本 理沙, 中山 宙, 松本 敏幸, 藤本 翔太, 佐藤 康成, 若山 清香, 若山 照彦, 内 村 有邦, 八木 健, 菅生 紀之
  • Organizer: 第44回日本分子生物学会年会
• [Presentation] Single-molecule imaging of CREB transcriptionfactor in human cortical-like neurons (2019)
  • Author(s): Yuri Atsumi, Noriyuki Sugo, Ryohei Iwata,Pierre Vanderhaeghen, Nobuhiko Yamamoto
  • Organizer: IBRO 2019
  • Int'l Joint Research
• [Presentation] Genome Stability by DNA polymerase β in Cortical Development (2018)
  • Author(s): 菅生紀之
  • Organizer: 第41回日本神経科学大会 シンポジウム「大脳皮質の形成と機能発現を担う遺伝子制御システムの新展開」
  • Invited
• [Presentation] Maintenance of genome stability by DNA polymerase β in necessary for postnatal development in cortical and hippocampal neurons (2018)
  • Author(s): Akiko Uyeda, Kohei Onishi, Teruyoshi Hirayama, Takeshi Yagi, Nobuhiko Yamamoto and Noriyuki Sugo
  • Organizer: 第41回日本神経科学大会
• [Presentation] DNA polymerase β is required for postnatal hippocampal development and function (2018)
  • Author(s): Akiko Uyeda, Kohei Onishi, Teruyoshi Hirayama, Satoko Hattori, Tsuyoshi Miyakawa, Takeshi Yagi, Nobuhiko Yamamoto and Noriyuki Sugo
  • Organizer: Cold Spring Harbor Asia meeting on Latest Advance in Development & Function of Neuronal Circuit
  • Int'l Joint Research
• [Presentation] Involvement of Polβ in postnatal neuronal development and function via active DNA demethylation (2018)
  • Author(s): Akiko Uyeda, Kohei Onishi, Teruyoshi Hirayama, Satoko Hattori, Tsuyoshi Miyakawa, Takeshi Yagi, Nobuhiko Yamamoto and Noriyuki Sugo
  • Organizer: 国立遺伝学研究所研究会「哺乳類脳の機能的神経回路の構築メカニズム」
• [Presentation] 神経活動依存的な転写因子動態の１分子蛍光イメージンング解析 (2017)
  • Author(s): 菅生紀之
  • Organizer: 第６９回日本細胞生物学会シンポジウム「クラップ＆ビルド・システムによる神経制御」（仙台）
  • Invited
• [Presentation] Genome stability by DNA polymerase β is Critical for Neuronal Differentiation in Cortical Development (2017)
  • Author(s): Noriyuki Sugo, Kohei Onishi, Akiko Uyeda, Mistuhiro Shida, Teruyoshi Hirayama, Takeshi Yagi, Nobuhiko Yamamoto
  • Organizer: Gene regulatory mechanisms in neural fate decisions (San Juan de Alicante, Spain)
  • Int'l Joint Research
• [Presentation] 大脳皮質神経細胞分化におけるDNAポリメラーゼβによるゲノム恒常性維持 (2017)
  • Author(s): 菅生紀之、大西公平、植田尭子、至田充宏、平山晃斉、八木健、山本亘彦
  • Organizer: 第３９回神経組織培養研究会（名古屋）
• [Presentation] 生後発達期の大脳皮質ニューロンにおけるDNAポリメラーゼβの役割 (2017)
  • Author(s): 植田尭子、菅生紀之、大西公平、平山晃斉、八木健、山本亘彦
  • Organizer: 第４０回日本神経科学大会（舞浜）
• [Presentation] iPS細胞を利用したヒト神経発生過程におけるDNAポリメラーゼ βの機能解析 (2017)
  • Author(s): 恒松大翔、菅生紀之、黒沢綾、足立典隆、山本亘彦
  • Organizer: 第４０回日本神経科学大会（舞浜）
• [Presentation] DNA polymerase β is required for active DNA demethylation in postnatal development of cortical neurons (2017)
  • Author(s): Akiko Uyeda, Noriyuki Sugo, Kohei Onishi, Teruyoshi Hirayama, Takeshi Yagi1 and Nobuhiko Yamamoto
  • Organizer: 第48回 生理研国際シンポジウム（岡崎）
• [Presentation] DNA polymerase β is involved in active DNA demethylation during postnatal development of cortical neurons (2017)
  • Author(s): Akiko Uyeda, Noriyuki Sugo, Kohei Onishi, Teruyoshi Hirayama, Takeshi Yagi1 and Nobuhiko Yamamoto
  • Organizer: 遺伝研研究会： 哺乳類脳の機能的神経回路の構築メカニズム（三島）
  • Int'l Joint Research
• [Book] Live-Cell Single-Molecule Imaging with Optogenetics Reveals Dynamics of a Neuronal Activity-Dependent Transcription Factor. In: Yamamoto N., Okada Y. (eds) Single Molecule Microscopy in Neurobiology. Neuromethods, vol 154 (2020)
  • Author(s): Kitagawa H., Sugo N., Yamamoto N
  • Total Pages: 20
  • Publisher: Humana, New York, NY
  • ISBN: 9781071605318
• [Remarks] 大阪大学大学院生命機能研究科
  • URL: http://www.fbs.osaka-u.ac.jp
• [Remarks] 脳の発生におけるゲノム安定維持の仕組みを解明
  • URL: http://www.fbs.osaka-u.ac.jp/jpn/events/achievement/yamamoto-20170801/
• [Remarks] 脳の発生におけるゲノム安定維持の仕組みを解明
  • URL: http://resou.osaka-u.ac.jp/ja/research/2017/20170821_1