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Acid inhibits TRPV4-mediated Ca(2)(+) influx in mouse esophageal epithelial cells Shikano M; Ueda T; Kamiya T; Ishida Y; Yamada T; Mizushima T; Shimura T; Mizoshita T; Tanida S; Kataoka H; Shimada S; Ugawa S; Joh TNeurogastroenterol Motil 2011[Nov]; 23 (11): 1020-8, e497BACKGROUND: The transient receptor potential vanilloid 4 (TRPV4), a thermo-sensitive stretch-activated cation channel, is expressed in the skin stratified squamous epithelium, contributing to the acquisition of barrier function. Similarly, functional TRPV4 may be located in the stratified squamous epithelial lining of the esophagus, being involved in the pathogenesis of gastroesophageal reflux disease (GERD). Here we investigated the expression of TRPV4 in the mouse esophageal epithelium. METHODS: TRPV4 expression at the mRNA and protein levels was examined by reverse transcription-polymerase chain reaction (RT-PCR), in situ hybridization, and immunohistochemistry. A calcium imaging technique and ATP assay were used to evaluate the functionality of TRPV4 in freshly isolated esophageal epithelial cells. KEY RESULTS: Transcripts and proteins encoding TRPV4 were colocalized in the basal and intermediate layers of the esophageal epithelium. Both 4alpha-phorbol 12,13- didecanoate (4alpha-PDD), a selective agonist for TRPV4, and hypo-osmolar solution (160 mOsm) elevated the intracellular calcium concentration ([Ca(2+) ](i) ) in a subset of the isolated cells (70%). These [Ca(2+) ](i) increases were potently inhibited by ruthenium red (RuR), a TRPV4 channel antagonist, and were suppressed by extracellular protons (pH 5.0). Finally, application of 4alpha-PDD evoked ATP release in primary esophageal epithelial cells. CONCLUSIONS & INFERENCES: Acid-sensitive TRPV4 channels were mainly expressed in the esophageal epithelial cells of the basal and intermediate layers. Direct exposure of TRPV4-expressing cells to gastric acid, as would occur in cases of GERD, could influence their cellular functions, possibly aggravating the disease state.|Acids/*metabolism[MESH]|Animals[MESH]|Calcium/*metabolism[MESH]|Epithelial Cells/cytology/*physiology[MESH]|Esophagus/*cytology[MESH]|HEK293 Cells[MESH]|Humans[MESH]|Male[MESH]|Mice[MESH]|Mice, Inbred C57BL[MESH]|TRPV Cation Channels/genetics/*metabolism[MESH] |
