Elucidation of the molecular basis of the ultra-early stage of endocytosis by high-resolution observation of cell membrane dynamics

2020 Fiscal Year Final Research Report

• Project/Area Number: 19K22506
• Research Category: Grant-in-Aid for Challenging Research (Exploratory)
• Allocation Type: Multi-year Fund
• Review Section: Medium-sized Section 48:Biomedical structure and function and related fields
• Research Institution: Hokkaido University
• Principal Investigator: Ohba Yusuke 北海道大学, 医学研究院, 教授 (30333503)
• Project Period (FY): 2019-06-28 – 2021-03-31
• Keywords: 細胞膜 / 高速分子間力顕微鏡 / 薄層遮光照明法 / 相関イメージング

Outline of Final Research Achievements

In this study, we utilized correlative imaging by hybrid microscopy combined with live-cell high-speed atomic force microscopy and fluorescence microscopy to observe and analyze the dynamics of cell membranes and molecules associated with clathrin-dependent endocytosis. We found that clathrin-coated pit formation induced by growth factor stimulation appears in a certain localized region of the plasma membrane and the morphology of the pit is not limited to circular (spherical), but can also be elliptical and divisive. Wey also found that Src is involved in such phenomena.

Free Research Field

細胞生理学

Academic Significance and Societal Importance of the Research Achievements

細胞内と細胞外環境を隔てる脂質二重膜であると同時に、細胞内外のコミュニケーションを司るインターフェースである細胞膜の3次元的なダイナミクスに関する知見はこれまで限られていた。本研究では、これまでの動態観察の分解能を10倍向上した高速AFMを用いることで、細胞と組織の活動や運命の決定に重要な細胞外からの刺激に応じた細胞膜の形態と成分の動的変化の解析を可能とし、かつその分子メカニズムの一端を明らかにした。