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 tension was present in adherent endothelial cells, we either cut or detached them from their basal surface by a microneedle. After cutting or detachment, the cells rapidly retracted. This retraction was prevented, however, if the CSK actin lattice was disrupted by cytochalasin D (Cyto 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 preexisting 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 adherent endothelial cells. Altered cell deformability was quantitated as the shear stiffness measured by magnetic twisting cytometry. When membrane 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 increased 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 mechanical changes in the forces distending the CSK but not to ATP- or Ca(2+)-dependent processes. Taken together, these results suggest preexisting CSK tension is a major determinant of cell deformability in adherent endothelial cells.
Pourati
J
J
Physiology Program, Harvard School of Public Health, Boston, Massachusetts 02215, USA.
Maniotis
A
A
Spiegel
D
D
Schaffer
J L
JL
Butler
J P
JP
Fredberg
J J
JJ
Ingber
D E
DE
Stamenovic
D
D
Wang
N
N
D002448:Cell Adhesion / Q000502:physiology; D003599:Cytoskeleton / Q000502:physiology*; D004548:Elasticity; D004727:Endothelium / Q000166:cytology* / Q000502:physiology*; D013314:Stress, Mechanical