Effects of fluid shear stress and cytoskeletal disruption on the actin cytoskeleton
AIM: To investigate the effects of fluid shear stress on cytoskeleton of osteoblasts, we examined the effects applying confocal laser scanning microscope and immunofluorescence microscopy. METHODS: The experiments were divided into 3 groups, Ⅰ group, fluid shear stress; Ⅱ group, fluid shear stress + cytochalasinD (CD); Ⅲ group, fluid shear stress + nocodazole (NO). 12 dyne/cm2 fluid shear stress was applied on the osteoblasts at four time points, and cells was marked by FITC-phalloidin and PI double label technique. Images were observed by confocal laser scanning microscope and immunofluorescence microscopy. RESULTS: After the application of fluid shear stress for 1 h at 12 dnye/cm2, F-actin filaments were organized into stress fibers that became thicker and more abundant than 0 min fluid shear stress cells. Shear stress was associated with significant increases of fluorescence intensity of F-actin. The minimum doses of nocodazole (microtubules) and cytochalasin D (actin filaments) partially disrupted the cytoskeleton. In cytochalasin D-treated cultures, F-actin filaments and the fluorescence intensity of F-actin significant decreased. While, F-actin slightly decreased in nocodazole treatment. CONCLUSION: These results indicate that the cytoskeleton, especially F-actin filaments, play a crucial role in cellular mechanotransduction.
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维普
