Chronic potassium restriction leads to active potassium reabsorption in the late distal nephron and collecting duct, segments known to express "gastric" H(+)-K(+)-adenosinetriphosphatase (H(+)-K(+)-ATPase) alpha-subunit mRNA. In this study, the cellular distribution and relative abundance of mRNA encoding this isoform were examined in kidneys of normal and potassium-deprived (2 wk) rats. In situ hybridization with isoform-specific cRNA probes demonstrated prominent expression of this gene in the connecting segment (CNT), entire collecting duct, and renal papillary surface epithelium in a comparable distribution in both groups of rats. Hypertrophy of the outer medullary collecting ducts in the inner stripe of potassium-restricted rats was observed. Competitive polymerase chain reaction analysis revealed twofold greater levels of gastric H(+)-K(+)-ATPase alpha-subunit mRNA (normalized to the level of beta-actin mRNA) in the cortex, but roughly comparable levels in the outer and inner medulla, of potassium-restricted rats compared with controls. These data suggest that chronic potassium restriction results in modestly enhanced renal cortical expression of the gastric H(+)-K(+)-ATPase alpha-subunit gene and that this isoform may participate in potassium conversation by the CNT and cortical collecting duct during potassium deprivation.