Rapid regulation of electrolyte absorption in sweat duct

Abstract

Even though the same Cl channel (CFTR) is common to certain fluid transport functions that are oppositely directed, i.e., secretion and absorption, only fluid secretion has clearly been shown to be acutely regulated. It is now clear that fluid secretion activated by beta-adrenergic stimulation is controlled by cAMP-mediated opening and closing of CFTR-Cl channels. Since the conductance of the human sweat duct is almost wholly due to CFTR-Cl conductance (CFTR-GCl), we sought to determine whether salt absorption via CFTR-Cl channels could also be subject to acute regulation in this purely absorptive epithelium. After alpha-toxin permeabilization, we found that addition of cAMP resulted in a large increase in Cl diffusion potentials across the apical membrane and a more than twofold increase in the average membrane conductance. Since the cAMP effects were dependent on Cl alone, not on Na, and since apical Cl conductance appears to be almost exclusively comprised of CFTR-GCl, we surmise that this form of electrolyte absorption like secretion is also subject to acute control through CFTR-GCl. Acute regulation of absorption involves both activation by phosphorylation (PKA) and inactivation by dephosphorylation (unknown endogenous phosphatase) of CFTR. Phosphorylation of CFTR was shown by the facts that CFTR-GCl could be activated by cAMP and inhibited by the kinase antagonist staurosporine, or by removal of either substrate ATP or Mg2+ cofactor. Inactivation of CFTR-GCl by endogenous phosphatase(s) was indicated by a spontaneous but reversible loss of CFTR-GCl upon removal of cAMP. Such loss of CFTR-GCl activity could be prevented either by application of phosphatase inhibitors or by using phosphatase-resistant ATP-gamma-S as substrate to phosphorylate CFTR. We surmise that absorptive function is subject to rapid regulation which can be switched "on" and "off" acutely by a control system that is common to both absorptive and secretory processes and that this control is crucial to switching between conductive and nonconductive transport mechanisms during salt absorption.