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Intracellular chloride activity and apical membrane chloride conductance inNecturus gallbladder

Research paper by J. Fernando Garcia-Diaz, Ayus Corcia, William McD Armstrong

Indexed on: 01 Jun '83Published on: 01 Jun '83Published in: The Journal of Membrane Biology



Abstract

Open-tip and Cl−-selective microelectrodes were used to study the effect of external pH on apical membrane potential (Va) and intracellular chloride activity (aCli) in epithelial cells ofNecturus gallbladder. Increasing the pH from 7.2 to 8.2 in the mucosal, the serosal, or in both bathing solutions simultaneously, hyperpolarizedVa (control value −60±5 mV) by about −6, −10 and −17 mV, respectively, but did not significantly change the transepithelial potential (VT=0.3±0.5 mV). Identical hyperpolarizations were recorded with Cl−-selective microelectrodes, even 40 min after changing external pH. Thus,aCli (12±2mm) remained essentially constant. The ratiofVa between the deflections inVa andVT produced by transepithelial current pulses, which is an approximate measure of the fractional resistance of the apical membrane, decreased when mucosal pH was increased, and increased when serosal pH was raised. The changes inVa andfVa are due, in part at least, to the known pH dependence of cell membrane K+ conductance (PK) in this tissue. The constancy ofaCli, despite significant increases inVa, indicates that cell membrane Cl− conductance (PCl) is virtually zero or decreases, with increased external pH, in a way that compensates for the increased driving force for Cl− exit. Experiments in which 90mm gluconate or 90mm methylsulfate were substituted for an equivalent amount of luminal Cl− did not provide any evidence for a significant contribution of Cl− ions, per se, to the emf or conductance of the apical membrane. They suggested, rather, a dependence of apical membrane cation permeability on luminal Cl− concentration. Since basolateral membranePCl is known to be very low, the insensitivity ofaCli toVa is the consequence of a negligible elctrodiffusive Cl− permeability at both cell membranes. Thus, overall, transcellular Cl− transport inNecturus gallbladder is, in large measure, effected by electroneutral processes.