Effects of aldosterone on the impedance properties of cultured renal amphibian epithelia

Abstract

The cultured renal amphibian cell line A6 has proven advantageous for studies of Na+ transport regulation. In the present study, the effects of aldosterone action on the transepithelial electrical properties of this epithelium were assessed. Specifically, the time course of aldosterone action was determined and the effects of chronic (10-18 day) aldosterone elevation were assessed using transepithelial equivalent circuit methods and impedance analysis techniques. Short-term (< 4 hr) exposure to aldosterone (0.1 microM) stimulated the amiloride-sensitive short-circuit current (Isc) by over twofold and increased the transepithelial conductance (GT) by approximately 12%. The increases in Isc and GT were maintained in epithelia subjected to chronic aldosterone exposure. In contrast to previous reports, paracellular resistance (Rj) was not altered by aldosterone. This difference may be related to the longer time of exposure or different basal Na+ transport rates in the present study. The apical membrane conductance was significantly increased for aldosterone-treated epithelia compared to aldosterone-depleted (i.e., serum-deprived) controls. Apical membrane area (capacitance) was not significantly affected. This finding is consistent with a higher density (number of channels per membrane area) of conducting Na+ channels in this membrane following aldosterone stimulation. Basolateral membrane properties were not significantly altered for aldosterone-treated tissues compared to serum-treated control tissues. In contrast, basolateral membrane-specific conductance (i.e., basolateral membrane conductance normalized to basolateral membrane capacitance) was significantly lower for serum-deprived epithelia than for serum-treated controls or aldosterone-treated tissues. The effects of chronic aldosterone exposure were also evaluated for the A6 subclonal cell line, 2F3. Similar to A6 epithelia, Isc was essentially doubled following aldosterone stimulation while Rj and cellular driving force (Ec) were not affected. Apical membrane conductances under control conditions for 2F3 epithelia were higher than those for A6, but were not significantly different from A6 following aldosterone exposure or serum deprivation. These findings suggest possible differences in the regulation of apical membrane Na+ channels for 2F3 and A6 epithelia.