Ca²⁺-sensing receptors in intestinal epithelium

Expression of Ca²⁺-sensing receptors (CAR) was demonstrated in several human intestinal epithelial cell lines (T84, HT-29, and Caco-2) and in rat intestinal epithelium by both reverse transcriptase-polymerase chain reaction (PCR) and Northern blotting of RNA. Restriction patterns of the PCR products were of the sizes predicted by the human and rat sequences. CaR agonists (Ca²⁺, poly-L-arginine, protamine) mediated an increase in intracellular Ca²⁺ in HT-2918-C1 cells (monitored by changes in fura 2 fluorescence), which was dependent on release from thapsigargin-sensitive stores. U-73122, an inhibitor of phosphatidylinositol-phospholipase C, eliminated the CaR agonist-mediated rise in intracellular Ca²⁺, whereas its inactive analog, U-73343, had no effect. Pertussis toxin pretreatment had no effect on CaR agonist-mediated modulation of intracellular Ca²⁺. Taken together, these studies demonstrate that CaR are expressed in intestinal epithelial cells and couple to mobilization of intracellular Ca²⁺. The presence of CaR in intestinal epithelial cells presents a new locus for investigations into the role(s) of extracellular Ca²⁺ in modulating intestinal epithelial cell differentiation and transepithelial Ca²⁺ transport.