Fluorescence single-molecule imaging of actin turnover and regulatory mechanisms

Naoki Watanabe

    Research output: Chapter in Book/Report/Conference proceedingChapter

    21 Citations (Scopus)

    Abstract

    Cells must rapidly remodel the actin filament network to achieve various cellular functions. Actin filament turnover is a dynamic process that plays crucial roles in cell adhesion, locomotion, cytokinesis, endocytosis, phagocytosis, tissue remodeling, etc., and is regulated by cell signaling cascades. Success in elucidating dynamic biological processes such as actin-based motility relies on the means enabling real time monitoring of the process. The invention of live-cell fluorescence single-molecule imaging has opened a window for direct viewing of various actin remodeling processes. In general, assembly and dissociation of actin and its regulators turned out to occur at the faster rates than previously estimated by biochemical and structural analyses. Cells undergo such fast continuous exchange of the components perhaps not only to drive actin remodeling but also to facilitate rapid response in many other cell mechanics and signaling cascades. This chapter describes how epifluorescence single-molecule imaging which visualizes deeper area than the TIRF microscopy is achieved in XTC cells, the currently best platform for this approach.

    Original languageEnglish
    Title of host publicationMethods in Enzymology
    PublisherAcademic Press Inc.
    Pages219-232
    Number of pages14
    DOIs
    Publication statusPublished - 2012

    Publication series

    NameMethods in Enzymology
    Volume505
    ISSN (Print)0076-6879
    ISSN (Electronic)1557-7988

    Keywords

    • abelson kinase
    • actin turnover
    • defective CMV promoter
    • drug perfusion
    • epi-fluorescence
    • flow cell
    • formin homology protein
    • photobleaching normalization
    • single-molecule imaging
    • XTC cell

    ASJC Scopus subject areas

    • Biochemistry
    • Molecular Biology

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