Molecular mechanisms of calcium signaling in the modulation of small intestinal ion transports and bicarbonate secretion

Abstract

Background and Purpose: Although Ca²⁺ signaling may stimulate small intestinal ion secretion, little is known about its critical role and the molecular mechanisms of Ca²⁺-mediated biological action.

Key Results: Activation of muscarinic receptors by carbachol(CCh) stimulated mouse duodenal I_sc, which was significantly inhibited in Ca²⁺-free serosal solution and by several selective store-operated Ca²⁺ channels(SOC) blockers added to the serosal side of duodenal tissues. Furthermore, we found that CRAC/Orai channels may represent the molecular candidate of SOC in intestinal epithelium. CCh increased intracellular Ca²⁺ but not cAMP, and Ca²⁺ signaling mediated duodenal Cl^- and HCO₃^- secretion in wild type mice but not in CFTR knockout mice. CCh induced duodenal ion secretion and stimulated PI3K/Akt activity in duodenal epithelium, all of which were inhibited by selective PI3K inhibitors with different structures. CCh-induced Ca²⁺ signaling also stimulated the phosphorylation of CFTR proteins and their trafficking to the plasma membrane of duodenal epithelial cells, which were inhibited again by selective PI3K inhibitors.

Materials and Methods: Functional, biochemical and morphological experiments were performed to examine ion secretion, PI3K/Akt and CFTR activity of mouse duodenal epithelium. Ca²⁺ imaging was performed on HT-29 cells.

Conclusions and Implications: Ca²⁺ signaling plays a critical role in intestinal ion secretion via CRAC/Orai-mediated SOCE mechanism on the serosal side of epithelium. We also demonstrated the molecular mechanisms of Ca²⁺ signaling in CFTR-mediated secretion via novel PI3K/Akt pathway. Our findings suggest new perspectives for drug targets to protect the upper GI tract and control liquid homeostasis in the small intestine.