IS CYTOSKELETAL TENSION A MAJOR DETERMINANT OF CELL DEFORMABILITY IN ADHERENT ENDOTHELIAL-CELLS

We tested the hypothesis that mechanical tension in the cytoskeleton ( CSK) is a major determinant of cell deformability. To confirm that ten sion was present in adherent endothelial cells, we either cut or detac hed them from their basal surface by a microneedle. After cutting or d etachment, the cells rapidly retracted. This retraction was prevented, however, if the CSK actin lattice was disrupted by cytochalasin D (Cy to D). These results confirmed that there was preexisting CSK tension in these cells and that the actin lattice was a primary stress-bearing component of the CSK. Second, to determine the extent to which that p reexisting CSK tension could alter cell deformability, we developed a stretchable cell culture membrane system to impose a rapid mechanical distension (and presumably a rapid increase in CSK tension) on adheren t endothelial cells. Altered cell deformability was quantitated as the shear stiffness measured by magnetic twisting cytometry. When membran e strain increased 2.5 or 5%, the cell stiffness increased 15 and 30%, respectively. Disruption of actin lattice with Cyto D abolished this stretch-induced increase in stiffness, demonstrating that the increase d stiffness depended on the integrity of the actin CSK. Permeabilizing the cells with saponin and washing away ATP and Ca2+ did not inhibit the stretch-induced stiffening of the cell. These results suggest that the stretch-induced stiffening was primarily due to the direct mechan ical changes in the forces distending the CSK but not to ATP-or Ca2+-d ependent processes. Taken together, these results suggest preexisting CSK tension is a major determinant of cell deformability in adherent e ndothelial cells.