A study of human neuronal circuit formation using iPS cells

• Project/Area Number: 15K14318
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Neurophysiology / General neuroscience
• Research Institution: Osaka University
• Principal Investigator: YAMAMOTO Nobuhiko 大阪大学, 生命機能研究科, 教授 (00191429)
• Project Period (FY): 2015-04-01 – 2017-03-31
• Project Status: Completed (Fiscal Year 2016)
• Budget Amount: ¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000); Fiscal Year 2016: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2015: ¥2,340,000 (Direct Cost: ¥1,800,000、Indirect Cost: ¥540,000)
• Keywords: iPS 細胞 / 神経細胞 / 細胞死 / 細胞分化 / polβ / iPS細胞 / DNA修復酵素 / 遺伝子発現 / CRISPR/Cas9 / 大脳皮質 / 軸索

Outline of Final Research Achievements

Our goal is to reveal characteristics of human neuronal differentiation and circuit formation using iPS cell technology. This study is aimed at making an experimental system to analyze the role of DNA polymerase β (polβ), by establishing the iPS cell line which lacks polβ. To establish the cell line, CRISPR/Cas9 system was utilized, and we could obtain the homozygote after drug selection. The obtained clone showed higher sensitivity than wild type clones. We further succeed in inducing neuronal stem cells and the neuronal cells.

Research Products

• [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 Issue: 1 Pages: 1-10
  • DOI: 10.1523/jneurosci.0943-16.2017
  • Peer Reviewed / Acknowledgement Compliant
• [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
  • ISBN: 9781493936656, 9781493936670
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Synapse-dependent and independent mechanisms of thalamocortical axon branching are regulated by neuronal activity (2016)
  • Author(s): Matsumoto, N., Hoshiko, M., Sugo, N., Fukazawa, Y. & Yamamoto, N.
  • Journal Title: Developmental Neurobiology Volume: 76 Issue: 3 Pages: 323-336
  • DOI: 10.1002/dneu.22317
  • Peer Reviewed
• [Journal Article] Single-molecule imaging reveals dynamics of CREB transcription factor bound to its target sequence (2015)
  • Author(s): Sugo, N., Morimatsu, M., Arai, Y., Kousoku, Y., Ohkuni, A., Nomura, T., Yanagida, T. & Yamamoto, N.
  • Journal Title: Scientific Report Volume: 5 Issue: 1 Pages: 10662-10662
  • DOI: 10.1038/srep10662
  • Peer Reviewed / Open Access
• [Presentation] KIT LIGAND ACTS AS A NEGATIVE REGULATOR FOR ACTIVITY-DEPENDENT THALAMOCORTICAL AXON BRANCHING (2016)
  • Author(s): Yasufumi Hayano, Takeshi Araki, Kensuke Sasaki, Yumi Miyasaka, Takuma Nakasone, Yoshio Hata, Nobuhiko Yamamoto
  • Organizer: Axon Guidance, synapse formation & regeneration
  • Place of Presentation: Cold Spring Harbor, NY, USA
  • Year and Date: 2016-09-20
  • Int'l Joint Research
• [Presentation] ARHGEF18 REGULATES AXON BRANCHING OF CORTICAL UPPER LAYER NEURONS (2016)
  • Author(s): Kensuke Sasaki, Kei Arimoto, Kento Kankawa, Chikayo Terada, Nobuhiko Yamamoto
  • Organizer: Axon Guidance, synapse formation & regeneration
  • Place of Presentation: Cold Spring Harbor, NY, USA
  • Year and Date: 2016-09-20
  • Int'l Joint Research
• [Presentation] Positive and negative regulation of activity-dependent thalamocortical axon branching (2016)
  • Author(s): Nobuhiko Yamamoto
  • Organizer: The 39th Annual Meeting of the Japan Neuroscience Society
  • Place of Presentation: 横浜
  • Year and Date: 2016-07-20
  • 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): Kohei Ohnishi, Noriyuki Sugo, Shunsuke Toyoda, Teruyoshi Hirayama, Takeshi Yagi, Nobuhiko Yamamoto
  • Organizer: The 39th Annual Meeting of the Japan Neuroscience Society
  • Place of Presentation: 横浜
  • Year and Date: 2016-07-20
  • Int'l Joint Research
• [Presentation] Involvement of DNA polymerase β in postnatal development of cortical neurons (2016)
  • Author(s): Akiko Uyeda, Noriyuki Sugo, Kohei Ohnishi, Shunsuke Toyoda, Teruyoshi Hirayama, Takeshi Yagi, Nobuhiko Yamamoto
  • Organizer: The 39th Annual Meeting of the Japan Neuroscience Society
  • Place of Presentation: 横浜
  • Year and Date: 2016-07-20
  • Int'l Joint Research
• [Presentation] Role of Kit ligand as a negative regulator in activity-dependent thalamocortical axon branching (2016)
  • Author(s): Yumi Miyasaka, Yasufumi Hayano, Takeshi Araki, Nobuhiko Yamamoto
  • Organizer: The 39th Annual Meeting of the Japan Neuroscience Society
  • Place of Presentation: 横浜
  • Year and Date: 2016-07-20
  • Int'l Joint Research
• [Presentation] Polβ deficiency increases DNA double-strand breaks in postnatal cortical neurons (2015)
  • Author(s): Uyeda, A., Onishi, K., Sugo, N., Toyoda S., Hiyayama, T., Yagi, T. & Yamamoto, N.
  • Organizer: 第38回日本分子生物学会
  • Place of Presentation: 神戸
  • Year and Date: 2015-12-01
• [Presentation] DNA polymerase β activity in neural progenitors is required for postmitotic neuronal survival in the developing cortex (2015)
  • Author(s): Onishi, k., Sugo, N., Toyoda, S., Hirayama, T., Yagi, T. & Yamamoto, N.
  • Organizer: Society for Neuroscience
  • Place of Presentation: Chicago, USA
  • Year and Date: 2015-10-17
  • Int'l Joint Research
• [Presentation] Activity-dependent dynamics of CREB in cortical neurons: A single-molecule imaging study (2015)
  • Author(s): Kitagawa, H., Sugo., N. & Yamamoto,
  • Organizer: Society for Neuroscience
  • Place of Presentation: Chicago, USA
  • Year and Date: 2015-10-17
  • Int'l Joint Research
• [Presentation] involvement of mitochondria in thalamocortical axon branching (2015)
  • Author(s): Matsumoto, N., Hoshiko, M. & Yamamoto, N.
  • Organizer: 第38回日本神経科学大会
  • Place of Presentation: 神戸
  • Year and Date: 2015-07-28
• [Presentation] Rho guanine nucleotide exchange factor Abr promotes axon branching in cortical layer 2/3 neurons DNA (2015)
  • Author(s): Sasaki, K., Arimoto, K., Kankawa, K., Terada, C. & Yamamoto, N.
  • Organizer: 第38回日本神経科学大会
  • Place of Presentation: 神戸
  • Year and Date: 2015-07-28
• [Remarks] 大阪大学大学院生命機能研究科 細胞分子神経生物学研究室 （山本亘彦研究室）
  • URL: http://www.fbs.osaka-u.ac.jp/labs/neurobiol/