Ouabain-sensitive and -insensitive K-ATPases in rat nephron: effect of K depletion

Because a ouabain-sensitive H-K-adenosinetriphosphatase (H-K-ATPase) has been identified recently in the amphibian bladder, we evaluated whether such an ATPase might exist also in the mammalian kidney, along with the ouabain-insensitive H-K-ATPase previously described in the collecting duct. For this purpose, we searched for an Na-independent, K-stimulated, ouabain- and Sch-28080-inhibitable ATPase activity in single segments of rat nephron. Ouabain-sensitive K-stimulated ATPase activity was detected in the absence of Na+ in rat proximal convoluted and straight tubules and in medullary and cortical thick ascending limbs of Henle's loop but not in collecting ducts. This K-ATPase differs from Na-K-ATPase by 1) its absence of requirement for Na, 2) its sensitivity to Sch-28080, 3) its higher sensitivity to ouabain, and 4) its absence in the collecting duct. It differs from the collecting duct H-K-ATPase by 1) its distribution along the nephron, 2) its sensitivity to ouabain, and 3) its lower sensitivity to Sch-28080. Furthermore, in rats fed a K-depleted diet for 2 wk, ouabain-sensitive K-ATPase activity was markedly reduced in both proximal tubules and thick ascending limbs, whereas collecting duct H-K-ATPase was upregulated.