Distal nephron fragments were isolated by Percoll density centrifugation from collagenase-dispersed rabbit kidney cortex and grown on collagen-coated permeable supports. By 5 days, transmonolayer pd (Vtm) was 3.2 ± 0.7 mV (apical side negative) and transmonolayer conductance (Gtm) = 3.5 ± 0.4mS·cm-2. Incubation of monolayers in the presence of aldosterone increased transport of sodium and potassium as well as net acid secretion. This was accompanied by an increase in the amiloride-sensitive equivalent Isc and a decrease in Gtm· Fluorescence microscopy was used to estimate the proportion of intercalated cells in the monolayers. Carbonic-anhydrase-rich cells accounted for 46%, peanut-agglutinin-positive cells accounted for 24%. Solute concentration differences between apical and basolateral fluids were also measured. After 24 h incubation, the concentrations (mM; mean ± SEM) for apical versus basolateral solutions, respectively, were: Na+ 123.48 ± 2.33 vs. 150.25 ± 0.53; Cl- 113.75 ± 1.24 vs. 129.04 ± 1.28; K+ 7.58 ± 0.52 vs. 3.99 ± 0.02; total CO2 9.25 ± 0.49 vs. 21.33 ± 0.45; pH was 7.03 ± 0.03 vs. 7.36 ± 0.01. Aldosterone stimulated Na+ and K+ transport as well as H+ secretion. This was accompanied by an increase in both the amiloride-sensitive and insensitive components of the equivalent short-circuit current. We found that the type of culture medium determined the expression of transport properties. Growth of monolayers in different media increased Na+ and K+ transport and either stimulated or inhibited acid secretion. Our findings are consistent with the idea that these cultures display both electrical and transport properties characteristic of cortical collecting duct.

Journal Section:
Original Paper
Keywords:
Collecting duct, Electrophysiology, Tissue culture, Sodium, Potassium, Principal cells, Intercalated cells
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© 1992 S. Karger AG, Basel
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1992
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