Mechanisms of morphological changes of mitochondria during neural differentiation and under disease condition by using cell-resealing technique

• Project/Area Number: 23657124
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Cell biology
• Research Institution: The University of Tokyo
• Principal Investigator: MURATA Masayuki 東京大学, 総合文化研究科, 教授 (50212254)
• Project Period (FY): 2011 – 2012
• Project Status: Completed (Fiscal Year 2013)
• Budget Amount: ¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000); Fiscal Year 2012: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000); Fiscal Year 2011: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
• Keywords: シグナル伝達 / 神経科学 / 細胞･組織 / セミインタクト細胞 / 細胞・組織 / 発生・分化

Research Abstract

We established a neural cell line N14.5-Tom5 cells, in which mitochondrial protein GFP-Tom5 is stably expressed. By using N14.5-Tom5, we found that activation of GSK3beta or Akt was involved in mitochondrial fission during neural differentiation, and identified the phosphorylation of Drp1, dynamin-related GTPase, by GSK3beta regulated the fission. Interestingly, the phosphorylation of Drp1 is reported be required for the mitochondrial fission at the onset of mitosis in mammalian cells. Next, we constructed model cells that replicated the conditions in neurons of Alzheimer's disease by using the resealing-cell technique and cytosol prepared from ApoE(-/-) mouse. As a result, we found that inactivity of Akt in the model cells caused the elongation of mitochondria and resulted in depressed mitochondrial function. Collectively, the persistent phosphorylation of both GSK3beta and Akt plays a crucial role for the maintaining normal mitochondrial morphology and function.

Research Products

• [Journal Article] PPARγ-induced PARylation promotes local DNA demethylation by production of 5-hydroxymethylcytosine (2013)
  • Author(s): Katsunori Fujiki, Akihiro Shinoda, Fumi Kano, Ryuichiro Sato, Katsuhiko Shirahige & Masayuki Murata
  • Journal Title: Nature Co㎜unications Volume: 4 Issue: 1 Pages: 2-262
  • DOI: 10.1038/ncomms3262
  • Peer Reviewed
• [Journal Article] The Semi-Intact Cell System and Methods for Cell Resealing : a Novel Systems Biology Tool to Elucidate Protein Networks with Spatio-Templioral Information (2013)
  • Author(s): Kano, F., Murata, M
  • Journal Title: Advances in Systems Biology Volume: 2(1) Pages: 6-14
  • Peer Reviewed
• [Journal Article] The semi-intact cell system and methods for cellresealing: a novel systems biology tool to elucidate protein networks with spatio-temporal information (2013)
  • Author(s): Kano,F., Murata, M
  • Journal Title: Advance in Systems Biology, Volume: 2 Pages: 6-14
  • Peer Reviewed
• [Journal Article] A Resealed-Cell System for Analyzing Pathogenic Intracellular Events : Perturbation of Endocytic Pathways under Diabetic Conditions. (2012)
  • Author(s): Fumi Kano, et al.
  • Journal Title: PLoS ONE. Volume: 7 Issue: 8 Pages: e44127-e44127
  • DOI: 10.1371/journal.pone.0044127
  • Peer Reviewed
• [Journal Article] 哺乳動物細胞ゴルジ体の細胞周期依存的ディスアッセンブリー (2012)
  • Author(s): 加納ふみ, 村田昌之
  • Journal Title: 生体の科学 Volume: 63巻5号 Pages: 404-407
• [Journal Article] 新しい機能を問われ出した小胞体-ゴルジ体中間区画 (ER-Golgi intermediate compartment : ERGIC) (2012)
  • Author(s): 菅原太一, 加納ふみ, 村田昌之
  • Journal Title: 生体の科学 Volume: 63巻5号 Pages: 424-425
• [Journal Article] セミインタクト細胞リシール法を用いた「病態モデル細胞」作製とその疾患研究への応用 (2012)
  • Author(s): 村田昌之, 加納ふみ
  • Journal Title: 化学と生物 Volume: Vol.50(No.7) Pages: 510-517
  • NAID: 10030820324
• [Journal Article] PKCd and e regulate the morphological integrity of the ER-Golgi intermediate compartment (ERGIC) but not the anterograde and retrograde transports via the Golgi apparatus (2012)
  • Author(s): Sugawara, T., Nakatsu, D., Kii, H., Maiya, N., Adachi, A., Yamamoto, A., Kano, F., Murata, M
  • Journal Title: Biochem. Biophys. Acta (Molecular Cell Research) Volume: 1823(4) Pages: 861-875
  • Peer Reviewed
• [Journal Article] Semi-intact cell system : Application to the nalysis of membrane trafficking beween the endoplasmic reticulum and the Golgi apparatus and of cell cycle-dependent changes in the morphology of these organelles (2012)
  • Author(s): Murata, M., Kano, F
  • Journal Title: in Crosstalk and Integration of Membrane Trafficking Pathways
• [Journal Article] KCδ and ε regulate the morphological integrity of the ER-Golgi intermediate compartment (ERGIC) but not the anterograde and retrograde transports via the Golgi apparatus. (2012)
  • Author(s): Sugawara, T., Nakatsu, D., Kii, H., Maiya, N., Adachi, A., Yamamoto, A., Kano, F., Murata, M.
  • Journal Title: Biochem. Biophys. Acta (Molecular Cell Research) Volume: 1823 Pages: 861-875
  • Peer Reviewed
• [Journal Article] 哺乳動物細胞ゴルジ体の細胞周期依存的ディスアッセンブリー (2012)
  • Author(s): 加納ふみ、村田昌之
  • Journal Title: 生体の科学 Volume: 63 Pages: 404-407
• [Journal Article] 新しい機能を問われ出した小胞体-ゴルジ体中間区画 (ER-Golgi intermediate compartment: ERGIC) (2012)
  • Author(s): 菅原太一、加納ふみ、村田昌之
  • Journal Title: 生体の科学 Volume: 63 Pages: 424-425
• [Journal Article] セミインタクト細胞リシール法を用いた「病態モデル細胞」作成とその疾患研究への応用 (2012)
  • Author(s): 村田昌之、加納ふみ
  • Journal Title: 化学と生物 Volume: 50 Pages: 510-517
• [Journal Article] PKCd and e regulate the morphological integrity of the ER-Golgi intermediate compartment (ERGIC) but not the anterograde and retrograde transports via the Golgi apparatus. (2012)
  • Author(s): Sugawara, T., Nakatsu, D., Kii, H., Maiya, N., Adachi, A., Yamamoto, A., Kano, F., Murata, M.
  • Journal Title: Biochem. Biophys. Acta (Molecular Cell Research) Volume: 1823 Pages: 861-875
  • Peer Reviewed
• [Journal Article] The small GTPase Cdc42 modulates the number of exocytosis-competent dense-core vesicles in PC12 cells. (2012)
  • Author(s): Sato, M.,Kitaguchi, T., Numano, R., Ikematsu, K., Kakeyama, M., Murata, M., Sato, K., Tsuboi, T.
  • Journal Title: Biochem. Biophys. Res. Commun. Volume: 420 Pages: 417-421
  • Peer Reviewed
• [Journal Article] Localization of mature Neprilysin in lipid rafts. (2011)
  • Author(s): Sato, K., Tanabe, C.,Yonemura, Y., Watahiki, H., Zhao, Y., Yagishita, S., Ebina, M., Suo, S., Futai, U., Murata, M., Ishiura, S.
  • Journal Title: J. Neurosci. Res. Volume: 90 Pages: 870-878
  • Peer Reviewed
• [Presentation] セミインタクト細胞リシール技術を用いた糖尿病モデル細胞の構築 (2013)
  • Author(s): 野口 誉之, 堀内 雄太, 中津 大貴, 加納 ふみ, 村田 昌之
  • Organizer: 第51回日本生物物理学会年会
  • Place of Presentation: 京都国際会議場
  • Year and Date: 2013-10-29
• [Presentation] Disease model cell system : a novel proteomics tool to elucidate protein networks with spatio-temporal information (2013)
  • Author(s): Fumi Kano, Daiki Nakatsu, Yoshiyuki Noguchi, Yuta Horiuchi, Masayuki Murata
  • Organizer: The 12th Human Proteome Organization Congress
  • Place of Presentation: Yokohama, Japan
  • Year and Date: 2013-09-16
• [Presentation] セミインタクト細胞リシール法を用いた病態モデル細胞の創成 (2013)
  • Author(s): 加納ふみ, 村田昌之
  • Organizer: 第65回日本細胞生物学会大会.シンポジウム「細胞への蛋白質導入技術と蛋白質イメージング技術」
  • Place of Presentation: ウインクあいち
  • Year and Date: 2013-06-19
• [Presentation] セミインタクト細胞リシール技術を用いた糖尿病モデル細胞の構築 : 病態依存的なエンドサイトーシス攪乱 (2013)
  • Author(s): 堀内雄太, 加納ふみ, 野口誉之, 村田昌之
  • Organizer: 第65回日本細胞生物学会大会(名古屋)
  • Year and Date: 2013-06-19
• [Presentation] セミインタクト細胞リシール技術を用いた糖尿病モデル細胞の構築 (2013)
  • Author(s): 野口誉之、堀内雄太、中津大貴、加納ふみ、村田昌之
  • Organizer: 第51回日本生物物理学会年会
  • Place of Presentation: 京都国際会議場(京都府)
• [Presentation] Disease model cell system: a novel proteomics tool to elucidate protein networks with spatio-temporal information. (2013)
  • Author(s): F. Kano, D. Nakatsu, Y. Noguchi, Y. Horiuchi, M. Murata.
  • Organizer: The 12th Human Proteome Organization Congress
  • Place of Presentation: パシフィコ横浜(神奈川県)
• [Presentation] セミインタクト細胞リシール技術を用いた糖尿病モデル細胞の構築：病態依存的なエンドサイトーシス攪乱 (2013)
  • Author(s): 堀内雄太、加納ふみ、野口誉之、村田昌之
  • Organizer: 第65回日本細胞生物学会大会
  • Place of Presentation: ウインクあいち(愛知県)
• [Presentation] セミインタクト細胞リシール法を用いた病態モデル細胞の創成 (2013)
  • Author(s): 加納ふみ、村田昌之
  • Organizer: 第65回日本細胞生物学会大会
  • Place of Presentation: ウインクあいち(愛知県)
• [Presentation] A Resealed-Cell System for Analyzing Pathogenic Intracellular Events : Insight into Signal Transduction Cascade in Hyperlipidemic or Diabetic Cells (2012)
  • Author(s): Masayuki Murata, Fumi Kano
  • Organizer: The 25th Annual and International Meeting of the Japanese Association for Animal Cell Technology (JAACT2012)
  • Place of Presentation: Nagoya Congress Center, Nagoya, Japan
  • Year and Date: 2012-11-27
• [Presentation] A resealed-cell system for analyzing pathogenic intracellular events: Insight into signal transduction cascade in hyperlipidemic or diabetic cells (2012)
  • Author(s): Masayuki Murata and Fumi Kano
  • Organizer: The 25th Annual and International Meeting of the Japanese Association for Animal Cell Technology
  • Place of Presentation: Nagoya Congress Center
  • Invited
• [Presentation] Establishment of semi-intact cell system and application of its resealing technique (2011)
  • Author(s): Masayuki Murata
  • Organizer: 第34回日本分子生物学会年会(招待講演)
  • Place of Presentation: パシフィコ横浜
• [Book] Semi-intact cell system: Application to the nalysis of membrane trafficking beween the endoplasmic reticulum and the Golgi apparatus and of cell cycle-dependent changes in the morphology of these organelles. in Crosstalk and Integration of Membrane Trafficking Pathways. (2012)
  • Author(s): Murata, M., Kano, F.
  • Publisher: INTECH