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Structured illumination microscopy reveals focal adhesions are composed of linear subunits

机译：结构照明显微镜显示粘着斑由线性亚基组成

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摘要

The ability to mechanically interact with the extracellular matrix is a fundamental feature of adherent eukaryotic cells. Cell-matrix adhesion in many cell types is mediated by protein complexes called focal adhesions (FAs). Recent progress in super resolution microscopy revealed FAs possess an internal organization, yet such methods do not enable observation of the formation and dynamics of their internal structure in living cells. Here, we combine structured illumination microscopy (SIM) with total internal reflection fluorescence microscopy (TIRF) to show that the proteins inside FA patches are distributed along elongated subunits, typically 300 +/- 100nm wide, separated by 400 +/- 100nm, and individually connected to actin cables. We further show that the formation and dynamics of these linear subunits are intimately linked to radial actin fiber formation and actomyosin contractility. We found FA growth to be the result of nucleation of new linear subunits and their coordinated elongation. Taken together, this study reveals that the basic units of mature focal adhesion are 300-nm-wide elongated, dynamic structures. We anticipate this ultrastructure to be relevant to investigation of the function of FAs and their behavior in response to mechanical stress. (c) 2015 Wiley Periodicals, Inc.
机译：与细胞外基质机械相互作用的能力是粘附的真核细胞的基本特征。在许多细胞类型中，细胞基质粘附是由称为焦点粘附（FA）的蛋白质复合物介导的。超分辨率显微镜的最新进展表明，FA具有内部组织，但这种方法无法观察活细胞中其内部结构的形成和动力学。在这里，我们将结构照明显微镜（SIM）与全内反射荧光显微镜（TIRF）结合使用，以显示FA斑块内的蛋白质沿细长的亚基分布，通常为300 +/- 100nm宽，相隔400 +/- 100nm，并且分别连接到肌动蛋白电缆。我们进一步表明，这些线性亚基的形成和动力学与放射状肌动蛋白纤维的形成和肌动球蛋白的收缩力密切相关。我们发现FA增长是新的线性亚基成核及其协调延伸的结果。两者合计，这项研究表明，成熟的粘着斑的基本单位是300纳米宽的细长的动态结构。我们预计这种超微结构与FAs的功能及其对机械应力的响应行为的研究有关。 （c）2015年威利期刊有限公司

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