Transient and steady-state effects of shear stress on endothelial cell adherens junctions

S Noria, DB Cowan, AI Gotlieb, BL Langille - Circulation research, 1999 - Am Heart Assoc
S Noria, DB Cowan, AI Gotlieb, BL Langille
Circulation research, 1999Am Heart Assoc
Endothelial cells exhibit profound changes in cell shape in response to altered shear stress
that may require disassembly/reassembly of adherens junction protein complexes that
mediate cell-cell adhesion. To test this hypothesis, we exposed confluent porcine aortic
endothelial cells to 15 dyne/cm2 of shear stress for 0, 8.5, 24, or 48 hours, using a parallel
plate flow chamber. Cells were fixed and stained with antibodies to vascular endothelial
(VE) cadherin, α-catenin, β-catenin, or plakoglobin. Under static conditions, staining for all …
Abstract
—Endothelial cells exhibit profound changes in cell shape in response to altered shear stress that may require disassembly/reassembly of adherens junction protein complexes that mediate cell-cell adhesion. To test this hypothesis, we exposed confluent porcine aortic endothelial cells to 15 dyne/cm2 of shear stress for 0, 8.5, 24, or 48 hours, using a parallel plate flow chamber. Cells were fixed and stained with antibodies to vascular endothelial (VE) cadherin, α-catenin, β-catenin, or plakoglobin. Under static conditions, staining for all proteins was intense and peripheral, forming a nearly continuous band around the cells at cell-cell junctions. After 8.5 hours of shear stress, staining was punctate and occurred only at sites of continuous cell attachment. After 24 or 48 hours of shear, staining for VE-cadherin, α-catenin, and β-catenin was intense and peripheral, forming a band of “dashes” (adherens plaques) that colocalized with the ends of stress fibers that inserted along the lateral membranes of cells. Staining for plakoglobin was not observed after 24 hours of shear stress, but returned after 48 hours. Western blot analysis indicated that protein levels of VE-cadherin, α-catenin, and plakoglobin decreased, whereas β-catenin levels increased after 8.5 hours of shear stress. As cell shape change reached completion (24 to 48 hours), all protein levels were upregulated except for plakoglobin, which remained below control levels. The partial disassembly of adherens junctions we have observed during shear induced changes in endothelial cell shape may have important implications for control of the endothelial permeability barrier and other aspects of endothelial cell function.
Am Heart Assoc