Membrane crosstalk in the mammalian proximal tubule during alterations in transepithelial sodium transport

Abstract

The present paper examines the effects of reduced transepithelial Na transport (JNa) on membrane electrophysiological parameters in proximal convoluted tubules and the possible role of cytosolic calcium concentration ([Ca]i) in the regulation of basolateral membrane K conductance (GK). When JNa was reduced by elimination of glucose and alanine and replacement of 100 mM sodium with N-methyl-D-glucamine from the luminal perfusate, basolateral membrane potential (VBL) hyperpolarized transiently by 12.6 mV and the ratio of apical to basolateral membrane resistance (RA/RBL) doubled. The apparent transference number for K at the basolateral membrane (GK/Gcell) decreased from 0.13 to 0.08 in the first 4 min following reductions in JNa. The elimination of Na-alanine and Na-glucose cotransport was responsible for the initial hyperpolarization and increase in RA/RBL, whereas the resultant decrease in the cellular concentrations of glucose and alanine, together with the reductions in GK, could elicit the secondary VBL depolarization. Measurement of [Ca]i with the fluorescent probe fura-2 during reductions in JNa revealed that [Ca]i increased by an average of 12%, a value very similar to the average reduction in cellular volume (13%) measured using morphometric techniques. The observation that [Ca]i increased while GK was decreasing is inconsistent with the effect of [Ca]i on putative basolateral Ca-activated K channel. We believe that [Ca]i changes passively (at least in the first few minutes) in response to a decrease in cell volume occurring as a consequence of reductions in JNa and that some as yet unidentified volume-sensitive mechanism is responsible for the regulation of GK.