Membrane transport parameters in frog corneal epithelium measured using impedance analysis techniques.

Abstract

Active Cl- transport in bullfrog corneal epithelium was studied using transepithelial impedance analysis methods, and direct-current (DC) measurements of membrane voltages and resistance ratios. The technique allows the estimation of the apical and basolateral membrane conductances, and the paracellular conductance, and does not rely on the use of membrane conductance-altering agents to obtain these measurements as was requisite in earlier DC equivalent-circuit analysis studies. In addition, the analysis results in estimates of the apical and basolateral membrane capacitances, and allows resolution of the paracellular conductance into properties of the tight junctions and lateral spaces. Membrane capacitances (proportional to areas) were used to estimate the specific conductances of the apical and basolateral membranes, as well as to evaluate coupling between the cell layers. We confirm results obtained from earlier studies: apical membrane conductance is proportional to the rate of active Cl- transport and is highly Cl- selective; intracellular Cl- activity is above electrochemical equilibrium, thereby providing a net driving force for apical membrane Cl- exit; the paracellular conductance is comparable to the transcellular conductance. We also found that: the paracellular conductance is composed of the series combination of the junctional conductance and a nonnegligible lateral space resistance; a small K+ conductance reported in the apical membrane may result from Cl- channels possessing a finite permeability to K+; the basolateral membrane areas is 36 times greater than the apical membrane area which is consistent with the notion of electrical coupling between the five to six cell layers of the epithelium; the specific conductance of the basolateral membrane is many times lower than that of the apical membrane; the net transport of Cl- is modulated primarily by changes in the conductance of the apical membrane and not by changes in the net electrochemical gradient resulting from opposite changes in the electrical and chemical gradients; the conductance of the basolateral membrane does not change with transport which implies that the net driving force for K+ exit increases with transport, possibly due to an increase in the intracellular K+ activity.