Abstract
Temporal behavior of cytoskeletons such as actin filaments, and microtubules and FAT (Focal Adhesion Targeting) domain was observed in sheared endothelial cells using gene transfection technique to understand their mechanical roles in endothelial cell remodeling. Plasmid encoding enhanced GFP-actin, GFP-tubulin and RFP-FAT domain were transfected into bovine aortic endothelial cells using a liposomal method. After gene transfection, cells were exposed to shear stress of 2 Pa using a parallel plate flow chamber and temporally observed by fluorescence microscopy. During the process of cell elongation and orientation to the direction of flow, dynamic process of the cytoskeletons and FAT domain was clearly observed, which was summarized as the following characteristic features: 1) formation of lamellipodia at upstream and downstream sides of shear flow, 2) shrinkage and depolymerization of actin filaments located at large angles to the direction of flow, 3) development of microtubule networks and appearance of FAT domain followed by formation of actin filaments between FAT domain. After disrupting microtubules with 2.5μg/ml nocodazole, development of lamellipodia was not observed. These results suggest that actin filaments would be involved in cell remodeling associated with FAT domain to form lamellipodia and microtubule networks would be required for appearance of FAT domain.
Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call
