Volume Reabsorption, Transepithelial Potential Differences, and Ionic Permeability Properties in Mammalian Superficial Proximal Straight Tubules

Abstract

This paper describes experiments designed to evaluate Na+ and Cl- transport in isolated proximal straight tubules from rabbit kidneys. When the perfusing solution was Krebs-Ringer buffer with 25 mM HCO3- (KRB) and the bath contained KRB plus 6% albumin, net volume reabsorption (Jv, nl min-1 mm-1 was -0.46 ± 0.03 (SEM); Ve, the spontaneous transepithelial potential difference, was -1.13 ± 0.05 mV, lumen negative. Both Jv, and Ve, were reduced to zero at 21°C or with 10-4 M ouabain, but Jv, was not HCO3- dependent. Net Na+ reabsorption, measured as the difference between 22Na+ fluxes, lumen to bath and bath to lumen, accounted quantitatively for volume reabsorption, assuming the latter to be an isotonic process, and was in agreement with the difference between lumen to bath 22Na+ fluxes during volume reabsorption and at zero volume flow. The observed flux ratio for Na+ was 1.46, and that predicted for a passive process was 0.99; thus, Na+ reabsorption was rationalized in terms of an active transport process. The Cl- concentration of tubular fluid rose from 113.6 to 132.3 mM during volume reabsorption. Since Ve, rose to +0.82 mV when tubules were perfused with 138.6 mM Cl- solutions, Ve may become positive when tubular fluid Cl- concentrations rise during volume reabsorption. The permeability coefficients PNa and PCl computed from tracer fluxes were, respectively, 0.23 x 10-4 and 0.73 x 10-4 cm s-1. A PNa/PCl ratio of 0.3 described NaCl dilution potentials at zero volume flow. The magnitudes of the potentials were the same for a given NaCl gradient in either direction and PNa/PCl was constant in the range 32–139 mM NaCl. We infer that the route of passive ion permeation was through symmetrical extracellular interfaces, presumably tight junctions, characterized by neutral polar sites in which electroneutrality is maintained by mobile counterions.