Gladkikh Aleena, Kovaleva Anastasia, Tvorogova Anna, Vorobjev Ivan A
Department of Cell Biology and Histology, School of Biology, M.V. Lomonosov Moscow State University, Moscow, Russia.
A.N. Belozersky Institute of Physico-Chemical Biology, M.V. Lomonosov Moscow State University, Moscow, Russia.
Methods Mol Biol. 2018;1745:205-218. doi: 10.1007/978-1-4939-7680-5_12.
Cell-extracellular matrix (ECM) adhesion is an important property of virtually all cells in multicellular organisms. Cell-ECM adhesion studies, therefore, are very significant both for biology and medicine. Over the last three decades, biomedical studies resulted in a tremendous advance in our understanding of the molecular basis and functions of cell-ECM adhesion. Based on morphological and molecular criteria, several different types of model cell-ECM adhesion structures including focal adhesions, focal complexes, fibrillar adhesions, podosomes, and three-dimensional matrix adhesions have been described. All the subcellular structures that mediate cell-ECM adhesion are quite heterogeneous, often varying in size, shape, distribution, dynamics, and, to a certain extent, molecular constituents. The morphological "plasticity" of cell-ECM adhesion perhaps reflects the needs of cells to sense, adapt, and respond to a variety of extracellular environments. In addition, cell type (e.g., differentiation status, oncogenic transformation, etc.) often exerts marked influence on the structure of cell-ECM adhesions. Although molecular, genetic, biochemical, and structural studies provide important maps or "snapshots" of cell-ECM adhesions, the area of research that is equally valuable is to study the heterogeneity of FA subpopulations within cells. Recently time-lapse observations on the FA dynamics become feasible, and behavior of individual FA gives additional information on cell-ECM interactions. Here we describe a robust method of labeling of FA using plasmids with fluorescent markers for paxillin and vinculin and quantifying the morphological and dynamical parameters of FA.
细胞与细胞外基质（ECM）的黏附是多细胞生物体中几乎所有细胞的一项重要特性。因此，细胞与ECM黏附的研究对生物学和医学都具有极其重要的意义。在过去三十年中，生物医学研究使我们对细胞与ECM黏附的分子基础和功能的理解取得了巨大进展。基于形态学和分子标准，已经描述了几种不同类型的细胞与ECM黏附结构模型，包括黏着斑、黏着斑复合体、纤维状黏附、足体和三维基质黏附。所有介导细胞与ECM黏附的亚细胞结构都具有很大的异质性，其大小、形状、分布、动态以及在一定程度上的分子组成常常各不相同。细胞与ECM黏附的形态“可塑性”或许反映了细胞感知、适应和响应各种细胞外环境的需求。此外，细胞类型（例如分化状态、致癌转化等）常常对细胞与ECM黏附的结构产生显著影响。尽管分子、遗传、生化和结构研究为细胞与ECM黏附提供了重要的图谱或“快照”，但同样有价值的研究领域是研究细胞内FA亚群的异质性。最近，对FA动态的延时观察变得可行，单个FA的行为为细胞与ECM相互作用提供了更多信息。在此，我们描述一种使用带有桩蛋白和纽蛋白荧光标记的质粒标记FA并量化FA形态和动态参数的可靠方法。
