TRPV4-mediated endothelial Ca2+ influx and vasodilation in response to shear stress

1 Department of Medicine, 2 Cardiovascular Center, 3 Department of Pediatrics, and 4 Department of Pathology, Medical College of Wisconsin, Milwaukee; 5 Children's Research Institute, Children's Hospital of Wisconsin, Milwaukee; 6 Veterans Administration Medical Center, Milwaukee, Wisconsin; and 7 Department of Pharmacology, Jichi Medical University, Tochigi, Japan Submitted September 9, 2009 ; accepted in final form December 1, 2009 The transient receptor potential vallinoid type 4 (TRPV4) channel has been implicated in the endothelial shear response and flow-mediated dilation, although the precise functions of this channel remain poorly understood. In the present study, we investigated the role of TRPV4 in shear stress-induced endothelial Ca 2+ entry and the potential link between this signaling response and relaxation of small resistance arteries. Using immunohistochemical analysis and RT-PCR, we detected strong expression of TRPV4 protein and mRNA in the endothelium in situ and endothelial cells freshly isolated from mouse small mesenteric arteries. The selective TRPV4 agonist GSK1016790A increased endothelial Ca 2+ and induced potent relaxation of small mesenteric arteries from wild-type (WT) but not TRPV4 –/– mice. Luminal flow elicited endothelium-dependent relaxations that involved both nitric oxide and EDHFs. Both nitric oxide and EDHF components of flow-mediated relaxation were markedly reduced in TRPV4 –/– mice compared with WT controls. Using a fura-2/Mn 2+ quenching assay, shear was observed to produce rapid Ca 2+ influx in endothelial cells, which was markedly inhibited by the TRPV4 channel blocker ruthenium red and TRPV4-specific short interfering RNA. Flow elicited a similar TRPV4-mediated Ca 2+ entry in HEK-293 cells transfected with TRPV4 channels but not in nontransfected cells. Collectively, these data indicate that TRPV4 may be a potential candidate of mechanosensitive channels in endothelial cells through which the shear stimulus is transduced into Ca 2+ signaling, leading to the release of endothelial relaxing factors and flow-mediated dilation of small resistance arteries. transient receptor potential; endothelium; endothelium-derived factors; calcium signaling Address for reprint requests and other correspondence: D. X. Zhang, Dept. of Medicine, Cardiovascular Center, Medical College of Wisconsin, 8701 Watertown Plank Rd., Milwaukee, WI 53226 (e-mail: xfzhang{at}mcw.edu ).