Role of Serosal TRPV4-Constituted SOCE Mechanism in Secretagogues-Stimulated Intestinal Epithelial Anion Secretion

As little is known about the role of calcium (CA²⁺) signaling mediating the small intestinal epithelial anion secretion, we aimed to study its regulatory role in secretagogue-stimulated duodenal anion secretion and the underlying molecular mechanisms. Therefore, intestinal anion secretion from native mouse duodenal epithelia was examined with Ussing chambers to monitor PGE₂-, 5-HT-, and CCh-induced short-circuit currents (I _sc ). PGE₂ (10 μM) and 5-HT (10 μM) induced mouse duodenal I _sc , markedly attenuated by serosal CA²⁺-free solution and selective blockers of store-operated CA²⁺ channels on the serosal side of the duodenum. Furthermore, PGE₂- and 5-HT-induced duodenal I _sc was also inhibited by ER CA²⁺ chelator TPEN. However, dantrolene, a selective blocker of ryanodine receptors, inhibited PGE₂-induced duodenal I _sc , while LiCl, an inhibitor of IP₃ production, inhibited 5-HT-induced I _sc . Moreover, duodenal I _sc response to the serosal applications of both PGE₂ and 5-HT was significantly attenuated in transient receptor potential vanilloid 4 (TRPV4) knockout mice. Finally, mucosal application of carbachol (100 μM) also induced duodenal I _sc via selective activation of muscarinic receptors, which was significantly inhibited in serosal CA²⁺-free solution but neither in mucosal CA²⁺-free solution nor by nifedipine. Therefore, the serosal TRPV4-constituted SOCE mechanism is likely universal for the most common and important secretagogues-induced and CA²⁺-dependent intestinal anion secretion. These findings will enhance our knowledge about gastrointestinal (G.I.) epithelial physiology and the associated G.I. diseases, such as diarrhea and constipation.