Recently, it was observed that the acetylcholine analogue carbachol induces a transient stimulation of an apical Cl− conductance in basolaterally depolarized rat distal colonic epithelium (Schultheiss et al., 2003). The further characterization of this conductance was the aim of the present study. All experiments were performed at basolaterally depolarized tissues (111.5 mmol·l−1 KCl buffer at the serosal side); in the absence of a K+ gradient, a Cl− current was driven across the apical membrane (107 mmol·l−1 K gluconate/4.5 mmol·l−1 KCl buffer on the mucosal side). Under these conditions, carbachol evoked an atropine-sensitive biphasic change in short-circuit current (I SC), consisting of a transient increase followed by a long-lasting decrease, suggesting a stimulation of apical Cl− conductance followed by an inhibition. This conductance was inhibited by SITS, but was resistant against glibenclamide, a blocker of CFTR. The carbachol-induced I SC was dependent on the presence of mucosal Ca2+. Ionomycin, a Ca2+ ionophore, mimicked the effect of carbachol. An antibody against bovine Ca2+-activated Cl− channel ClCa1 stained rat colonic epithelial cells both at the cell membrane as well as intracellularly, suggesting that the action of Ca2+ may be caused by a stimulation of a ClCa-type anion channel. The activation of apical Cl− conductance by carbachol was resistant against any blockers of the phospholipase C/IP3/protein kinase C pathway tested (e.g., U-73122, 2-ABP, Li+, staurosporine), but was inhibited by the NO-synthase blocker L-NNA. Vice versa, NO-donating compounds such as GEA 3162 or sodium nitroprusside evoked a transient increase of I SC. Consequently, NO seems to be involved in the transient stimulation of apical Ca2+-dependent Cl− conductance after muscarinic receptor stimulation.
The Journal of Membrane Biology – Springer Journals
Published: Jan 1, 2005
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