In Vitro Actin Cytoskeleton Organization During Endothelial Monolayer Integrity Formation And Regulation Revealed By Super-resolution Microscopyстатья
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Дата последнего поиска статьи во внешних источниках: 10 июня 2020 г.
Аннотация:Endothelial cells have a flattened shape and, tightly adjacent to each other, form a layer of cells lining the inner surface of blood vessels. The endothelium plays a barrier role and regulates vascular tone and vascular wall permeability. However, the endothelial monolayer is sensitive to the effects of various physical and chemical stimuli, both in physiological and pathological states. The barrier function efficiency is dependent on the interaction of cell cytoskeleton structures and adhesive (cell-cell and cell-substrate contacts) components. All three types of cytoskeletal structures (microtubules, actin and intermediate filaments) operate coordinately through various linkers and mediators, contributing to the strengthening or weakening of the endothelial barrier. Initially, barrier function and dysfunction researches were focused on the actin system of the endothelial cells. Actin structures are an essential component for cell contraction. In endothelial cells, both in vivo and in vitro, two actin isoforms are present: non-muscle β- and γ-cytoplasmic actin. In this study, double immunofluorescent labeling and super-resolution microscopy (Structural Illuminational Microscopy, SIM), as well as correlation analysis, were used to study the relative position of cytoskeleton components consisting of different actin isoforms during endothelial monolayer integrity formation and regulation. The results of the study demonstrate significant differences in actin structures intracellular localization – β-actin is found mainly in stress fibers and ring bundles, and the extensive actin network of the cell cytoplasm consists of γ-actin. In the area of lamellipodia, near to complete colocalization of cytoplasmic β- and γ-actin structures is often observed, but simultaneously, in the same endotheliocyte, colocalization is not observed in the zone of stable cell edges and in cell central regions surrounding the cell nucleus. Relative localization of β- and γ-actin components changes during the formation of endothelial cell monolayer. Similar changes are observed during endothelial barrier dysfunction development resulting of experimental microtubule- depolymerizing treatments. The reported study was supported by Russian Foundation for Basic Research (project number 18 29 09082) and Moscow State University Development Program (PNR 5.13).