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Imaging myosin-driven stress fiber contraction with molecular resolution by high-speed AFM

Research Project

Project/Area Number 20H03218
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Review Section Basic Section 43040:Biophysics-related
Research InstitutionKanazawa University

Principal Investigator

Franz Clemens  金沢大学, ナノ生命科学研究所, 准教授 (50837664)

Project Period (FY) 2020-04-01 – 2023-03-31
Project Status Completed (Fiscal Year 2023)
Budget Amount *help
¥18,070,000 (Direct Cost: ¥13,900,000、Indirect Cost: ¥4,170,000)
Fiscal Year 2022: ¥4,030,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥930,000)
Fiscal Year 2021: ¥4,030,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥930,000)
Fiscal Year 2020: ¥10,010,000 (Direct Cost: ¥7,700,000、Indirect Cost: ¥2,310,000)
KeywordsAFM / SICM / myosin / actin / stress fiber / contractility / HS-AFM / Myosin / Actin / High-speed AFM / Bioimaging / SPM / cytoskeleton / Stress Fibers
Outline of Research at the Start

This research projects investigates the structure and function of contractile stress fibers, which have been compared to intracellular "muscles". Understanding how these structures contract will help us to better understand how cell can move, change shape, or divide.

Outline of Final Research Achievements

By combining cell de-roofing and HS-AFM imaging, this project established a method to image contracting actin stress fibers (SFs) to near molecular resolution within native cellular environments. Achieving these goals required (1) the development of suitable de-roofing methods, (2) establishment of an ultrawide scanner system for large resolution HS-AFM imaging in combination with fluorescence microscopy, and (3) developing an optimized AFM tip design for imaging highly corrugated intracellular structures. Ultrawide HS-AFM scanning generated the first molecular-resolution maps of entire native SFs, providing novel insight into SF ultrastructure. Furthermore, arrangement and conformational changes of individual non-muscle myosin II motors during SF contraction could be visualized for the first time. In parallel, SICM imaging was also adapted for intracellular imaging in de-roofed cells for the first time, providing additional structural and mechanical insight into SF contraction.

Academic Significance and Societal Importance of the Research Achievements

アクチンストレスファイバー(SF)の収縮は、マトリックス接着、遊走、メカノセンシングなど、多くの細胞プロセスに影響を与える一方、SF機能の誤制御は、アテローム性動脈硬化症、骨粗鬆症、がんなどの疾患の原因となりうる。このプロジェクトで得られたSFの超微細構造とSF収縮を駆動する分子メカニズムに関する新たな知見は、SFの正常および異常機能についての理解を広げるものである。

Report

(2 results)
  • 2023 Final Research Report ( PDF )
  • 2020 Annual Research Report
  • Research Products

    (6 results)

All 2021 2020

All Journal Article (3 results) (of which Int'l Joint Research: 3 results,  Peer Reviewed: 2 results,  Open Access: 2 results) Presentation (3 results) (of which Invited: 2 results)

  • [Journal Article] An ultra-wide scanner for large-area high-speed atomic force microscopy with megapixel resolution2021

    • Author(s)
      Marchesi Arin、Umeda Kenichi、Komekawa Takumi、Matsubara Takeru、Flechsig Holger、Ando Toshio、Watanabe Shinji、Kodera Noriyuki、Franz Clemens M.
    • Journal Title

      Scientific Reports

      Volume: 11 Issue: 1 Pages: 13003-13003

    • DOI

      10.1038/s41598-021-92365-y

    • Related Report
      2020 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Atomic reconstruction of biomolecular structures from AFM images and quantitative validation of experimental data using simulated AFM scanning2021

    • Author(s)
      Amyot Romain、Marchesi Arin、Franz Clemens M、Casuso Ignacio、Flechsig Holger
    • Journal Title

      bioRxiv

      Volume: 0 Pages: 0-0

    • DOI

      10.1101/2021.06.27.450070

    • Related Report
      2020 Annual Research Report
    • Open Access / Int'l Joint Research
  • [Journal Article] Integrin α2β1 as a negative regulator of the laminin receptors α6β1 and α6β42021

    • Author(s)
      Dao Lu、Blaue Carina、Franz Clemens M.
    • Journal Title

      Micron

      Volume: 148 Pages: 103106-103106

    • DOI

      10.1016/j.micron.2021.103106

    • Related Report
      2020 Annual Research Report
    • Peer Reviewed / Int'l Joint Research
  • [Presentation] Quantitating cell adhesion and mechanics using atomic force microscopy2021

    • Author(s)
      Clemens M. Franz
    • Organizer
      Open Lab Bruker Webinar
    • Related Report
      2020 Annual Research Report
    • Invited
  • [Presentation] Applications for HS-AFM in Cell Biology2021

    • Author(s)
      Clemens M. Franz
    • Organizer
      European Phys2Biomed Progress Meeting
    • Related Report
      2020 Annual Research Report
  • [Presentation] Imaging Myosin II-driven Stress Fiber Contraction in De-roofed Cells by High-Speed AFM2020

    • Author(s)
      Clemens M. Franz
    • Organizer
      8th Multifrequency AFM Conference, Madrid, Spain
    • Related Report
      2020 Annual Research Report
    • Invited

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Published: 2020-04-28   Modified: 2025-01-30  

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