Flow-mediated regulation of G-protein expression in cocultured vascular smooth muscle and endothelial cells

Arteriosclerosis, Thrombosis, and Vascular Biology
E M RedmondJ V Sitzmann

Abstract

G-proteins have been implicated in the transduction of a number of flow-induced responses. We determined whether flow can modulate vascular endothelial or smooth muscle cell G-protein signaling. By use of a perfused transcapillary coculture system that permits the chronic exposure of cultured endothelial cells (ECs) and smooth muscle cells (SMCs) to physiological shear stresses, cocultures were exposed to stepwise increases in flow up to (1) 2 mL/min (low flow: 0.5 dyne/cm2), or (2) 44 mL/min (high flow: 15 dyne/cm2) and maintained for 72 hours before SMCs and ECs were harvested separately. Using Western blot analysis, EC Gi alpha3 expression was significantly increased (41+/-2.9%) by high-flow conditions compared with low-flow. The changes in G-protein expression were associated with a significant increase in endothelial nitric oxide synthase (eNOS) activity, elevated prostacyclin levels in the perfusing media, increased pertussis toxin-catalyzed ADP ribosylation of Gi alpha substrates, and enhanced agonist-stimulated GTPase activity in cocultured ECs. In contrast, high flow induced a significant decrease in Gi alpha1-2 expression (57 5%) in SMCs cocultured with ECs, an effect that was endothelium dependent, inhibited by indom...Continue Reading

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Citations

Aug 23, 2001·Pharmacology & Therapeutics·P A CahillJ V Sitzmann
Oct 5, 2001·Journal of the American College of Cardiology·M ImamuraC Tei
Oct 23, 2012·American Journal of Physiology. Lung Cellular and Molecular Physiology·Devon ScottWei Tan
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Jul 15, 2006·Endothelium : Journal of Endothelial Cell Research·Han Qin WangZong Lai Jiang
Aug 31, 2002·Endothelium : Journal of Endothelial Cell Research·Shailesh Y DesaiDamir Janigro
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Jan 3, 2001·Medicinal Research Reviews·C E HillS L Sandow

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