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Spatiotemporal imaging of membrane morphology and protein localization during endocytic process

Research Project

Project/Area Number 16K14722
Research Category

Grant-in-Aid for Challenging Exploratory Research

Allocation TypeMulti-year Fund
Research Field Cell biology
Research InstitutionKyoto University

Principal Investigator

Shigehiro Yoshimura  京都大学, 生命科学研究科, 准教授 (90346106)

Co-Investigator(Renkei-kenkyūsha) ADACHI Taiji  京都大学, ウイルス・再生医科学研究所, 教授 (40243323)
Project Period (FY) 2016-04-01 – 2018-03-31
Project Status Completed (Fiscal Year 2017)
Budget Amount *help
¥3,770,000 (Direct Cost: ¥2,900,000、Indirect Cost: ¥870,000)
Fiscal Year 2017: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
Fiscal Year 2016: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
Keywordsエンドサイトーシス / シグナル伝達 / 細胞骨格 / アクチン / クラスリン / 原子間力顕微鏡 / ナノイメージング / endocytosis / clathrin / membrane dynamics / actin / signal transduction / 高速原子間力顕微鏡 / 表層アクチン / 細胞膜 / 細胞膜受容体 / メカノバイオロジー / 情報伝達
Outline of Final Research Achievements

We established hybrid time-lapse imaging system by combining high-speed atomic force microscopy and confocal laser-scanning microscopy to simultaneously image membrane morphology and protein localization during clathrin-mediated endocytosis. Spatiotemporal assembly of proteins such as clathrin, dynamin, epsin and actin, were successfully revealed together with morphological changes of the membrane. The inhibition of actin dynamics affected assembly, maturation and closing steps, suggesting important roles of cortical actin dynamics in multiples steps of endocytic process. At the closing step, most of the pits were covered by a membrane swelling, which is induced by a quick burst of actin polymerization, and then irreversibly closed, whereas some pits were closed without these motions. By combining membrane model and dynamic actin polymerization model, the mechanism of the pit closure was analyzed.

Report

(3 results)
  • 2017 Annual Research Report   Final Research Report ( PDF )
  • 2016 Research-status Report
  • Research Products

    (16 results)

All 2018 2017 2016 Other

All Journal Article (5 results) (of which Peer Reviewed: 4 results,  Open Access: 1 results,  Acknowledgement Compliant: 1 results) Presentation (10 results) (of which Int'l Joint Research: 4 results,  Invited: 4 results) Remarks (1 results)

  • [Journal Article] Morphological changes of plasma membrane and protein assembly during clathrin-mediated endocytosis2018

    • Author(s)
      A. Yoshida, N. Sakai, Y. Uekusa, Y. Imaoka, Y. Itagaki, Y. Suzuki and S.H. Yoshimura
    • Journal Title

      PLOS Biology

      Volume: 印刷中

    • NAID

      120006491910

    • Related Report
      2017 Annual Research Report
  • [Journal Article] In vivo dynamics of the cortical actin network revealed by fast-scanning atomic force microscopy2017

    • Author(s)
      Zhang Yanshu、Yoshida Aiko、Sakai Nobuaki、Uekusa Yoshitsugu、Kumeta Masahiro、Yoshimura Shige H.
    • Journal Title

      Microscopy

      Volume: 66 Issue: 4 Pages: 272-282

    • DOI

      10.1093/jmicro/dfx015

    • Related Report
      2017 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] In vivo dynamics of the cortical actin network revealed by fast-scanning atomic force microscopy2017

    • Author(s)
      65.Y Zhan, A Yoshida, N. Sakai, Y Uekusa, M. Kumeta and S.H. Yoshimura
    • Journal Title

      Microscopy

      Volume: 印刷中

    • NAID

      120006401728

    • Related Report
      2016 Research-status Report
    • Peer Reviewed / Acknowledgement Compliant
  • [Journal Article] HEAT repeats - versatile arrays of amphiphilic helices working in crowded environments?2016

    • Author(s)
      S.H. Yoshimura and T. Hirano
    • Journal Title

      J. Cell Sci.

      Volume: 129 Issue: 21 Pages: 3963-3970

    • DOI

      10.1242/jcs.185710

    • Related Report
      2016 Research-status Report
    • Peer Reviewed
  • [Journal Article] Identification of a nucleoporin358-specific RNA aptamer for use as a nucleus-targeting liposomal delivery system.2016

    • Author(s)
      G. Shrivastava, M. Hyodo, S.H. Yoshimura, H. Akita and H. Harashima
    • Journal Title

      Nuc. Acid Ther.

      Volume: 26 Issue: 5 Pages: 286-298

    • DOI

      10.1089/nat.2016.0604

    • Related Report
      2016 Research-status Report
    • Peer Reviewed
  • [Presentation] Morphological analysis of clathrin-mediated endocytotic process by fast-scanning atomic force microscope2017

    • Author(s)
      YOSHIDA A, ZHAN Y, ITAGAKI Y, KUMETA M, SUZUKI Y, SAKAI N, UEKUSA Y, YOSHIMURA SH
    • Organizer
      第58回日本植物生理学会年会
    • Place of Presentation
      鹿児島
    • Year and Date
      2017-03-17
    • Related Report
      2016 Research-status Report
    • Invited
  • [Presentation] Involvement of actin dynamics in endocytotic process revealed by fast-scanning atomic force microscope2017

    • Author(s)
      YOSHIDA A, SAKAI N, UEKUSA Y, ZHAN Y, KUMETA M, YOSHIMURA SH
    • Organizer
      Biophysical Society 61st Annual Meeting
    • Place of Presentation
      New Orleans, USA
    • Year and Date
      2017-02-15
    • Related Report
      2016 Research-status Report
    • Int'l Joint Research
  • [Presentation] Live-cell analysis of actin dynamics by fast-scanning atomic force microscope2017

    • Author(s)
      YOSHIMURA SH
    • Organizer
      Nuclear Actin in the Regulation of Transcription and Nuclear Structure
    • Place of Presentation
      Wakayama, Japan
    • Year and Date
      2017-01-26
    • Related Report
      2016 Research-status Report
    • Int'l Joint Research / Invited
  • [Presentation] Live-cell imaging of actin dynamics in cortex and lamellipodium by high-speed atomic force microscopy2017

    • Author(s)
      Y. Itagaki, Y. Zhang, A. Yoshida, N. Sakai, Y. Uekusa, M. Kumeta and S.H. Yoshimura
    • Organizer
      日本生物物理学会第55回年会
    • Related Report
      2017 Annual Research Report
    • Invited
  • [Presentation] Live-cell analysis of actin network by high-speed atomic force microscopy2017

    • Author(s)
      Y. Itagaki, Y. Zhang, A. Yoshida, M. Kumeta, and S.H. Yoshimura
    • Organizer
      日本生物物理学会第55回年会
    • Related Report
      2017 Annual Research Report
  • [Presentation] Live-cell imaging by high-speed atomic force microscopy revealed protein and plasma membrane dynamics during endocytic process2017

    • Author(s)
      S.H. Yoshimura
    • Organizer
      EMBO/EMBL Symposium: Seeing is Believing - Imaging the process of Life
    • Related Report
      2017 Annual Research Report
    • Int'l Joint Research
  • [Presentation] 高速原子間力顕微鏡を用いた細胞膜動態のライブセルイメージング2017

    • Author(s)
      吉田藍子、酒井信明、植草良嗣、粂田昌宏、吉村成弘、大場雄介
    • Organizer
      第97回北海道医学大会生理系分科会
    • Related Report
      2017 Annual Research Report
  • [Presentation] Role of cortical actin dynamics in endocytic process revealed by fast-scanning atomic force microscope2016

    • Author(s)
      YOSHIDA A, ZHAN Y, SUZUKI Y, ITAGAKI, Y, SAKAI N, UEKUSA Y, KUMETA M, YOSHIMURA SH
    • Organizer
      IGER International Symposium on Now in actin study
    • Place of Presentation
      Nagoya. Japan
    • Year and Date
      2016-12-12
    • Related Report
      2016 Research-status Report
    • Int'l Joint Research
  • [Presentation] 高速原子間力顕微鏡を用いた生体膜と皮質アクチンネットワーク動態のライブセルイメージング2016

    • Author(s)
      吉田藍子,酒井信明,植草良嗣、張雁書,粂田昌宏,伊藤修一,吉村成弘
    • Organizer
      第39回日本分子生物学会年会
    • Place of Presentation
      横浜
    • Year and Date
      2016-11-30
    • Related Report
      2016 Research-status Report
  • [Presentation] 高速原子間力顕微鏡による生体膜動態のライブセルイメージング2016

    • Author(s)
      吉田藍子,酒井信明,植草良嗣,粂田昌宏,吉村成弘
    • Organizer
      生体膜と薬物の相互作用シンポジウム
    • Place of Presentation
      名古屋
    • Year and Date
      2016-11-17
    • Related Report
      2016 Research-status Report
    • Invited
  • [Remarks] 研究室のホームページに研究成果を掲載

    • URL

      http://www.chrom.lif.kyoto-u.ac.jp/

    • Related Report
      2017 Annual Research Report

URL: 

Published: 2016-04-21   Modified: 2019-03-29  

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