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Am J Physiol Cell Physiol 275: C484-C495, 1998;
0363-6143/98 $5.00
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Vol. 275, Issue 2, C484-C495, August 1998

Distinct Ca2+- and cAMP-dependent anion conductances in the apical membrane of polarized T84 cells

Didier Merlin1,4, Lianwei Jiang2,4, Gregg R. Strohmeier1,4, Asma Nusrat1,4, Seth L. Alper2,4, Wayne I. Lencer3,4, and James L. Madara1,4

1 Division of Gastrointestinal Pathology, Department of Pathology, Brigham and Women's Hospital, 2 Molecular Medicine and Renal Units, Beth Israel Deaconess Medical Center, 3 Combined Program in Pediatric Gastroenterology and Nutrition, Department of Medicine, Children's Hospital Medical Center, 4 Harvard Medical School and Harvard Digestive Diseases Center, Boston, Massachusetts 02115

Monolayers of the human colonic epithelial cell line T84 exhibit electrogenic Cl- secretion in response to the Ca2+ agonist thapsigargin and to the cAMP agonist forskolin. To evaluate directly the regulation of apical Cl- conductance by these two agonists, we have utilized amphotericin B to permeabilize selectively the basolateral membranes of T84 cell monolayers. We find that apical anion conductance is stimulated by both forskolin and thapsigargin but that these conductances are differentially sensitive to the anion channel blocker DIDS. DIDS inhibits thapsigargin-stimulated responses completely but forskolin responses only partially. Furthermore, the apical membrane anion conductances elicited by these two agonists differ in anion selectivity (for thapsigargin, I- > Cl-; for forskolin, Cl- > I-). However, the DIDS-sensitive component of the forskolin-induced conductance response exhibits anion selectivity similar to that induced by thapsigargin (I- > Cl-). Thus forskolin-induced apical anion conductance comprises at least two components, one of which has features in common with that elicited by thapsigargin.

intestinal epithelium; thapsigargin; forskolin; chloride channels; amphotericin B; human intestinal cell line T84


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