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cotransport: role of Cl and
F-actin
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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