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Am J Physiol Renal Physiol 283: F923-F933, 2002. First published June 26, 2002; doi:10.1152/ajprenal.00015.2002
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Vol. 283, Issue 5, F923-F933, November 2002

Sodium transporter abundance profiling in kidney: effect of spironolactone

Jakob Nielsen1,2, Tae-Hwan Kwon2, Shyama Masilamani1, Kathleen Beutler1, Henrik Hager2, Søren Nielsen2, and Mark A. Knepper1

1 Laboratory of Kidney and Electrolyte Metabolism, National Heart, Lung and Blood Institute, National Institutes of Health, Bethesda, Maryland 20892-1603; and 2 The Water and Salt Research Center, University of Aarhus, DK-8000 Aarhus C, Denmark

Renal tubule profiling studies were carried out to investigate the long-term effects of administration of spironolactone, a mineralocorticoid receptor antagonist, on abundances of the major Na transporter and Na channel proteins along the rat renal tubule. Oral administration of spironolactone for 7 days to NaCl-restricted rats did not significantly alter abundances of Na transporters expressed proximal to the macula densa, while substantially decreasing the abundances of the thiazide-sensitive Na-Cl cotransporter (NCC), the alpha -subunit of the amiloride-sensitive epithelial Na channel (ENaC), and the 70-kDa form of the gamma -subunit of ENaC. A dependency of NCC expression on aldosterone was confirmed by showing increased NCC expression in response to aldosterone infusion in adrenalectomized rats. Immunoperoxidase labeling of ENaC in renal cortex confirmed that dietary NaCl restriction causes a redistribution of ENaC to the apical domain of connecting tubule cells and showed that high-dose spironolactone administration does not block this apical redistribution. In contrast, spironolactone completely blocked the increase in alpha -ENaC abundance in response to dietary NaCl restriction. We conclude that the protein abundances of NCC, alpha -ENaC, and the 70-kDa form of gamma -ENaC are regulated via the classical mineralocorticoid receptor, but the subcellular redistribution of ENaC in response to dietary NaCl restriction is not prevented by blockade of the mineralocorticoid receptor.

aldosterone; collecting duct; distal convoluted tubule


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