title: Aldosterone-mediated regulation of ENaC alpha, beta, and gamma subunit proteins in rat kidney.
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 alphaENaC protein without increasing the abundance of the beta and gamma subunits. Thus, alphaENaC 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 gammaENaC 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.affiliation: Laboratory of Kidney and Electrolyte Metabolism, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, Maryland 20892, USA.
authors.forename: S
authors.initials: S
authors.lastname: Masilamani
authors.forename: G H
authors.initials: GH
authors.lastname: Kim
authors.forename: C
authors.initials: C
authors.lastname: Mitchell
authors.forename: J B
authors.initials: JB
authors.lastname: Wade
authors.forename: M A
authors.initials: MA
authors.lastname: Knepper
mesh_terms: D000450:Aldosterone / Q000097:blood / Q000494:pharmacology / Q000502:physiology*; D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies; D000918:Antibody Specificity; D053503:Epithelial Sodium Channels; D000939:Epitopes / Q000737:chemistry / Q000276:immunology; D005786:Gene Expression Regulation / Q000187:drug effects / Q000502:physiology*; D015151:Immunoblotting; D007150:Immunohistochemistry; D007685:Kidney Tubules, Collecting / Q000187:drug effects / Q000502:physiology*; D046911:Macromolecular Substances; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000737:chemistry; D011817:Rabbits; D051381:Rats; D015222:Sodium Channels / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics*
chemical_list: D000906:Antibodies; D053503:Epithelial Sodium Channels; D000939:Epitopes; D046911:Macromolecular Substances; D010446:Peptide Fragments; D015222:Sodium Channels; D000450:Aldosterone