Cellular Biology |
From the Institute for Medicine and Engineering (B.P.H., P.F.D.), Department of Pathology and Laboratory Medicine (P.F.D.), and Department of Bioengineering (B.P.H., P.F.D.), University of Pennsylvania, Philadelphia, Pa; Department of Cell and Molecular Biology (R.D.G.), Northwestern University Medical School, Chicago, Ill.
Correspondence to Peter F. Davies, PhD, Institute for Medicine and Engineering, University of Pennsylvania, 1010 Vagelos Research Labs, 3340 Smith Walk, Philadelphia, PA 19104-6383. E-mail pfd{at}pobox.upenn.edu
AbstractHemodynamic shear stress at the endothelial cell surface induces acute and chronic intracellular responses that regulate vessel wall biology. The cytoskeleton is implicated by acting both as a direct connector to local surface deformation and as a distribution network for mechanical forces throughout the cell; however, direct observation and measurement of its position during flow have only recently become possible. In this study, we directly demonstrate rapid deformation of the intermediate filament (IF) network in living endothelial cells subjected to changes in hemodynamic shear stress. Time-lapse optical sectioning and deconvolution microscopy were performed within the first 3 minutes after the introduction of flow (shear stress, 12 dyn/cm2). Spatial and temporal dynamics of green fluorescent proteinvimentin IFs in confluent endothelial cells were analyzed. The imposition of shear stress significantly increased the variability of IF movement throughout the cell in the x-, y-, and z-directions compared with the constitutive dynamics noted in the absence of flow. Acute polymerization and depolymerization of the IF network were absent. The magnitude and direction of flow-induced IF displacement were heterogeneous at the subcellular level. These qualitative and quantitative data demonstrate that shear stress acting at the luminal surface of the endothelium results in rapid deformation of a stable IF network.
Key Words: mechanotransduction endothelium green fluorescent protein
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