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1 The Whitaker Institute of Biomedical Engineering and Department of Bioengineering, University of California, San Diego, La Jolla, California 92093-0427; 2 Center for Biomedical Engineering and Department of Mechanical Engineering, City College of New York, New York, New York 10031
Blood flow-associated
shear stress may modulate cellular processes through its action on the
plasma membrane. We quantified the spatial and temporal aspects of the
effects of shear stress (
) on the lipid fluidity of
1,1'-dihexadecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate [DiIC16(13)]-stained plasma membranes
of bovine aortic endothelial cells in a flow chamber. A confocal
microscope was used to determine the DiI diffusion coefficient
(D) by fluorescence recovery after photobleaching on cells
under static conditions, after a step- of 10 or 20 dyn/cm2, and after the cessation of . The method
allowed the measurements of D on the upstream and downstream
sides of the cell taken midway between the respective cell borders and
the nucleus. In <10 s after a step- of 10 dyn/cm2,
D showed an upstream increase and a downstream decrease, and both changes disappeared rapidly. There was a secondary, larger increase in upstream D, which reached a peak at 7 min and decreased thereafter, despite the maintenance of .
D returned to near control values within 5 s after
cessation of . Downstream D showed little secondary
changes throughout the 10-min shearing, as well as after its cessation.
Further investigations into the early phase, with simultaneous
measurements of upstream and downstream D, confirmed that a
step- of 10 dyn/cm2 elicited a rapid (5-s) but transient
increase in upstream D and a concurrent decrease in
downstream D, yielding a significant difference between the
two sites. A step- of 20 dyn/cm2 caused D to
increase at both sites at 5 s, but by 30 s and 1 min the
upstream D became significantly higher than the downstream D. These results demonstrate shear-induced changes in
membrane fluidity that are time dependent and spatially heterogeneous. These changes in membrane fluidity may have important implications in
shear-induced membrane protein modulation.
mechanotransduction; membrane fluidity; fluorescence recovery after photobleaching; cholesterol; alcohol
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