Abstract
Dynamic behavior of actin filaments and focal adhesion targeting (FAT) was observed in endothelial cells during exposure to fluid flow. Cultured bovine aortic endothelial cells were co-transfected with GFP-actin and RFP-FAT using a liposomal method. Fluid shear stress of 2Pa was then applied to confluent endothlial cell monolayer using a parallel-plate flow chamber. During experiments, fluorescence images were observed every 10min up to 50min by a laser scanning microscope. Before exposure to shear stress, a number of small FAT clusters were observed and the distal tip of actin filaments was associated with the FAT clusters. After exposure to shear stress, FAT clusters were rearranged together with actin filaments, showing prominent features such as appearance/disappearance, aggregation and sliding. These results indicated that the present method should be very useful for understanding the mechanism of mechanical responses of endothelial cells.
