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lüll The TRPV4 cation channel mediates stretch-evoked Ca2+ influx and ATP release in primary urothelial cell cultures Mochizuki T; Sokabe T; Araki I; Fujishita K; Shibasaki K; Uchida K; Naruse K; Koizumi S; Takeda M; Tominaga MJ Biol Chem 2009[Aug]; 284 (32): 21257-64Transient receptor potential channels have recently been implicated in physiological functions in a urogenital system. In this study, we investigated the role of transient receptor potential vanilloid 4 (TRPV4) channels in a stretch sensing mechanism in mouse primary urothelial cell cultures. The selective TRPV4 agonist, 4alpha-phorbol 12,13-didecanoate (4alpha-PDD) evoked Ca(2+) influx in wild-type (WT) urothelial cells, but not in TRPV4-deficient (TRPV4KO) cells. We established a cell-stretch system to investigate stretch-evoked changes in intracellular Ca(2+) concentration and ATP release. Stretch stimulation evoked intracellular Ca(2+) increases in a stretch speed- and distance-dependent manner in WT and TRPV4KO cells. In TRPV4KO urothelial cells, however, the intracellular Ca(2+) increase in response to stretch stimulation was significantly attenuated compared with that in WT cells. Stretch-evoked Ca(2+) increases in WT urothelium were partially reduced in the presence of ruthenium red, a broad TRP channel blocker, whereas that in TRPV4KO cells did not show such reduction. Potent ATP release occurred following stretch stimulation or 4alpha-PDD administration in WT urothelial cells, which was dramatically suppressed in TRPV4KO cells. Stretch-dependent ATP release was almost completely eliminated in the presence of ruthenium red or in the absence of extracellular Ca(2+). These results suggest that TRPV4 senses distension of the bladder urothelium, which is converted to an ATP signal in the micturition reflex pathway during urine storage.|*Urination[MESH]|Adenosine Triphosphate/*metabolism[MESH]|Animals[MESH]|Calcium Signaling[MESH]|Calcium/*metabolism[MESH]|Male[MESH]|Mice[MESH]|Mice, Inbred C57BL[MESH]|Mice, Transgenic[MESH]|Models, Biological[MESH]|Muscle, Smooth/metabolism/*pathology[MESH]|Phorbol Esters/metabolism[MESH]|Ruthenium Red/pharmacology[MESH]|TRPV Cation Channels/metabolism/*physiology[MESH]|Urothelium/*metabolism[MESH] |