Podocytes Respond to Mechanical Stress In Vitro

突触素 细胞骨架 生物物理学 肌动蛋白 化学 波形蛋白 足细胞 应力纤维 细胞生物学 肌动蛋白细胞骨架 中间灯丝 细胞 生物 生物化学 免疫组织化学 蛋白尿 免疫学 内分泌学
作者
Nicole Endlich,Kai R. Kress,Jochen Reiser,Dietmar Uttenweiler,Wilhelm Kriz,Peter Mündel,Karlhans Endlich
出处
期刊:Journal of The American Society of Nephrology 卷期号:12 (3): 413-422 被引量:278
标识
DOI:10.1681/asn.v123413
摘要

Abstract. Glomerular capillary pressure is thought to affect the structure and function of glomerular cells. However, it is unknown whether podocytes are intrinsically sensitive to mechanical forces. In the present study, differentiated mouse podocytes were cultured on flexible silicone membranes. Biaxial cyclic stress (0.5 Hz and 5% linear strain) was applied to the membranes for up to 3 d. Mechanical stress reduced the size of podocyte cell bodies, and processes became thin and elongated. Podocytes did not align in the inhomogeneous force field. Whereas the network of microtubules and that of the intermediate filament vimentin exhibited no major changes, mechanical stress induced a reversible reorganization of the actin cytoskeleton: transversal stress fibers (SF) disappeared and radial SF that were connected to an actin-rich center (ARC) formed. Epithelial and fibroblast cell lines did not exhibit a comparable stress-induced reorganization of the F-actin. Confocal and electron microscopy revealed an ellipsoidal and dense filamentous structure of the ARC. Myosin II, α-actinin, and the podocyte-specific protein synaptopodin were present in radial SF, but, opposite to F-actin, they were not enriched in the ARC. The formation of the ARC and of radial SF in response to mechanical stress was inhibited by nonspecific blockade of Ca 2+ influx with Ni 2+ (1 mM), by Rho kinase inhibition with Y-27632 (10 μM), but not by inhibition of stretch-activated cation channels with Gd 3+ (50 μM). In summary, mechanical stress induces a unique reorganization of the actin cytoskeleton in podocytes, featuring radial SF and an ARC, which differ in protein composition. The F-actin reorganization in response to mechanical stress depends on Ca 2+ influx and Rho kinase. The present study provides the first direct evidence that podocytes are mechanosensitive.

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