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Am J Physiol Gastrointest Liver Physiol 283: G240-G250, 2002. First published March 20, 2002; doi:10.1152/ajpgi.00500.2001
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Vol. 283, Issue 1, G240-G250, July 2002

Expression of calcium-sensing receptor in rat colonic epithelium: evidence for modulation of fluid secretion

Sam X. Cheng1, Masahiro Okuda1, Amy E. Hall1, John P. Geibel1,2, and Steven C. Hebert1

1 Department of Cellular and Molecular Physiology and 2 Department of Surgery, Yale University, New Haven, Connecticut 06520-8026

The calcium-sensing receptor (CaSR) is activated by extracellular calcium (Ca<UP><SUB>o</SUB><SUP>2+</SUP></UP>) and mediates many of the known effects of extracellular divalent minerals on body cells. Both surface and crypt cells express CaSR transcripts and protein on both apical and basolateral surfaces. Raising Ca<UP><SUB>o</SUB><SUP>2+</SUP></UP> elicited increases in intracellular calcium (Ca<UP><SUB>i</SUB><SUP>2+</SUP></UP>) in both surface and crypt cells with an EC50 of 2 mM. The Ca<UP><SUB>o</SUB><SUP>2+</SUP></UP>-induced increase in Ca<UP><SUB>i</SUB><SUP>2+</SUP></UP> was associated with increases in inositol 1,4,5-trisphosphate and eliminated by U-73129, an inhibitor of phosphatidylinositol-phospholipase C, as well as by thapsigargin. Other CaSR agonists, Gd3+ and neomycin, mimicked these Ca<UP><SUB>o</SUB><SUP>2+</SUP></UP>-induced responses. Both luminal and bath Ca<UP><SUB>o</SUB><SUP>2+</SUP></UP>, Gd3+, and neomycin induced increases in Ca<UP><SUB>i</SUB><SUP>2+</SUP></UP> in isolated perfused crypts. The stimulatory effect of forskolin on net fluid secretion in perfused crypts was abolished by increasing Ca<UP><SUB>o</SUB><SUP>2+</SUP></UP> in either luminal or bath perfusates. Thus both apical and basolateral CaSR on crypt cells are functional and provide pathways modulating net intestinal fluid transport that may have important implications for the prevention and treatment of certain diarrheal diseases associated with elevated cAMP.

calcium-sensing receptor; intracellular calcium; net fluid movement; isolated perfused colonic crypt


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