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J Membr Biol. 2013 Apr;246(4):271-85. doi: 10.1007/s00232-013-9528-9. Epub 2013 Feb 9.

STIM1-regulated Ca2+ influx across the apical and the basolateral membrane in colonic epithelium.

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  • 1Institute for Veterinary Physiology and Biochemistry, Justus-Liebig-University, Giessen, Germany.

Abstract

In nonexcitable cells, store-operated Ca(2+) entry is the most important pathway for influx of extracellular Ca(2+) serving as a second messenger in the cytoplasm. The present study investigated the expression, localization and polar distribution of two key components of store-operated Ca(2+) entry identified, e.g., in lymphocytes or epithelial cell lines-STIM1 (stromal interacting molecule 1), working as a Ca(2+) sensor in the endoplasmic reticulum, and Orai1, working as the (or part of the) store-operated Ca(2+) channel in the plasma membrane-in a native intestinal epithelium, i.e., rat colon. Immunohistochemical investigations revealed expression of STIM1 and Orai1 in the rat colonic epithelium. Ca(2+) store depletion led to a translocation of STIM1 both to the basolateral as well as to the apical cell pole as observed by confocal microscopy. A Ca(2+) depletion/repletion protocol was used in Ussing chamber experiments to investigate the contribution of basolateral and apical store-operated Ca(2+) entry to the induction of anion secretion. These experiments revealed that Ca(2+)-dependent anion secretion was induced not only by basolateral Ca(2+) repletion but also, to a lesser extent, by apical Ca(2+) repletion. Both responses were suppressed by La(3+). The effect of basolateral Ca(2+) repletion was significantly inhibited by brefeldin A, a blocker of vesicular transport from the endoplasmic reticulum to the Golgi apparatus. In a final series of experiments, fura-2-loaded HT29/B6 cells were used. A carbachol-induced increase in the cytosolic Ca(2+) concentration was significantly reduced when cells were pretreated with siRNA against STIM1. In conclusion, these results demonstrate that STIM1 as a key component of intracellular Ca(2+) signaling is expressed by rat colonic epithelium and is involved in the regulation not only of basolateral but also of apical Ca(2+) influx.

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