Water and urea transport in renal microvillus membrane vesicles

Abstract

Light scattering was used to measure the water and urea permeability of brush border membrane vesicles (BBMV) isolated from rabbit renal cortex. In stop-flow experiments, exposure of BBMV to a 200 mM inwardly directed mannitol gradient gave a monophasic time course of decreasing BBMV volume corresponding to an osmotic water permeability (Pf) of 1.1 +/- 0.1 X 10(-2) cm/s at 37 degrees C. The temperature dependence of Pf was biphasic with delta H = 2 kcal/mol for T less than 33 degrees C and delta H = 14 kcal/mol for T greater than 33 degrees C. A 200 mM inwardly directed urea gradient gave a biphasic time course of BBMV volume due to rapid water efflux (approximately 50 ms) followed by slower urea influx (1-5 s) with urea permeability (Purea) of 2.4 +/- 0.2 X 10(-6) cm/s. Preincubation of BBMV with increasing [urea] reversibly inhibited both urea flux (Kd = 1,200 mM) and thiourea flux (Kd = 370 mM) according to a single-site inhibition model, suggesting a saturable urea carrier. Comparison of BBMV Pf and Purea with proximal tubule transepithelial water and urea transport rates suggests that the permeability of the tubular apical membrane (BBMV) is high enough to support a transcellular route for both osmotic water and urea transport.