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Aldosterone-mediated regulation of ENaC α, β, and γ subunit proteins in rat kidney
Shyama Masilamani, … , James B. Wade, Mark A. Knepper
Shyama Masilamani, … , James B. Wade, Mark A. Knepper
Published October 1, 1999
Citation Information: J Clin Invest. 1999;104(7):R19-R23. https://doi.org/10.1172/JCI7840.
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Aldosterone-mediated regulation of ENaC α, β, and γ subunit proteins in rat kidney

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Abstract

Aldosterone stimulates sodium transport in the renal collecting duct by activating the epithelial sodium channel (ENaC). To investigate the basis of this effect, we have developed a novel set of rabbit polyclonal antibodies to the 3 subunits of ENaC and have determined the abundance and distribution of ENaC subunits in the principal cells of the rat renal collecting duct. Elevated circulating aldosterone (due to either dietary NaCl restriction or aldosterone infusion) markedly increased the abundance of αENaC protein without increasing the abundance of the β and γ subunits. Thus, αENaC is selectively induced by aldosterone. In addition, immunofluorescence immunolocalization showed a striking redistribution in ENaC labeling to the apical region of the collecting duct principal cells. Finally, aldosterone induced a shift in molecular weight of γENaC from 85 kDa to 70 kDa, consistent with physiological proteolytic clipping of the extracellular loop as postulated previously. Thus, at the protein level, the response of ENaC to aldosterone stimulation is heterogenous, with both quantitative and qualitative changes that can explain observed increases in ENaC-mediated sodium transport.

Authors

Shyama Masilamani, Gheun-Ho Kim, Carter Mitchell, James B. Wade, Mark A. Knepper

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Figure 2

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Effect of dietary NaCl restriction for 10 days. (a) Semiquantitative imm...
Effect of dietary NaCl restriction for 10 days. (a) Semiquantitative immunoblots for ENaC subunits and other major renal sodium transporters. For each blot, each lane was loaded with a homogenate from a different rat (n = 6 for both control and sodium-restricted rats). Preliminary Coomassie-stained gels demonstrated equality of loading among lanes. Asterisks mark statistically significant increases. (b) Deglycosylation with PNGase. SDS-solubilized samples from control rats and NaCl-restricted rats were incubated with PNGase F (200 U/40 μg protein; New England Biolabs Inc., Beverly, Massachusetts, USA) or vehicle.

Copyright © 2022 American Society for Clinical Investigation
ISSN: 0021-9738 (print), 1558-8238 (online)

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