An inwardly-directed sodium-amino acid cotransporter influences steady-state cell volume in slices of rat renal papilla incubated in hyperosmotic media

The effect of a neutral amino acid, 2-aminoisobutyric acid (AIB) on steady state cell volume has been examined in rat renal papillary slices incubated in hyperosmotic media (2,000 mosmol/kg H2O) containing high concentrations of NaCl and urea (thus imitating papillary interstitial fluid in the intact kidney during antidiuresis). Volumes were significantly increased (P less than 0.001) when external AIB was raised from 0.1 to 10 mmol/l. Na+-dependent AIB uptake occurred, and there were net increases in cell contents of Na+ and Cl-. Replacement of Na+ by Li+, but not by other cations, did not influence the effect of AIB concentration on cell volume, but this was abolished when Cl- was replaced by other anions. The effect of AIB was abolished by diphenylamine-2-carboxylate (10(-3) mmol/l), bumetanide (at 1 mmol/l but not 10(-2) mmol/l) and by N,N'-dicyclohexylcarbodiimide (0.5 mmol/l), but not by amiloride (1 mmol/l) or 4-acetamido-4'-iso-thiocyanato-stilbene-2,2'-disulphonic acid (1 mmol/l), and was enhanced by the presence of Ba2+ or quinine (1 mmol/l). The findings are interpreted in terms of an inwardly-directed Na+-amino acid cotransporter, which determines steady-state volume, requires simultaneous entry of Cl- through conductive pathways, and whose effects on cell volume are moderated by K+ efflux through volume-sensitive K+ channels.