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Am J Physiol Cell Physiol 274: C697-C706, 1998;
0363-6143/98 $5.00
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Vol. 274, Issue 3, C697-C706, March 1998

Osmotic regulation of intestinal epithelial Na+-K+-Clminus cotransport: role of Clminus and F-actin

Jeffrey B. Matthews, Jeremy A. Smith, Edward C. Mun, and Jason K. Sicklick

Department of Surgery, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, Massachusetts 02215

Previous data indicate that adenosine 3',5'-cyclic monophosphate activates the epithelial basolateral Na+-K+-Cl- cotransporter in microfilament-dependent fashion in part by direct action but also in response to apical Cl- loss (due to cell shrinkage or decreased intracellular Cl-). To further address the actin dependence of Na+-K+-Cl- cotransport, human epithelial T84 monolayers were exposed to anisotonicity, and isotopic flux analysis was performed. Na+-K+-Cl- cotransport was activated by hypertonicity induced by added mannitol but not added NaCl. Cotransport was also markedly activated by hypotonic stress, a response that appeared to be due in part to reduction of extracellular Cl- concentration and also to activation of K+ and Cl- efflux pathways. Stabilization of actin with phalloidin blunted cotransporter activation by hypotonicity and abolished hypotonic activation of K+ and Cl- efflux. However, phalloidin did not prevent activation of cotransport by hypertonicity or isosmotic reduction of extracellular Cl-. Conversely, hypertonic but not hypotonic activation was attenuated by the microfilament disassembler cytochalasin D. The results emphasize the complex interrelationship among intracellular Cl- activity, cell volume, and the actin cytoskeleton in the regulation of epithelial Cl- transport.

cytoskeleton; microfilament; bumetanide; NKCC1; BSC2


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