MECHANISMS OF EGF-INDUCED STIMULATION OF SODIUM-REABSORPTION BY ALVEOLAR EPITHELIAL-CELLS

We investigated the effects of epidermal growth factor (FGF) on active Na+ absorption by alveolar epithelium. Rat alveolar epithelial cells (AEC) were isolated and cultivated in serum-free medium on tissue cult ure-treated polycarbonate filters. mRNA for rat epithelial Na+ channel (rENaC) alpha-, beta-, and gamma-subunits and Na+ pump alpha(1)- and beta(1)-subunits were detected in day 4 monolayers by Northern analysi s and were unchanged in abundance in day 5 monolayers in the absence o f EGF. Monolayers cultivated in the presence of EGF (20 ng/ml) for 24 h from day 4 to day 5 showed an increase in both alpha(1) and beta(1) Na+ pump subunit mRNA but no increase in rENaC subunit mRNA. EGF-treat ed monolayers showed parallel increases in Na+ pump alpha(1)- and beta (1)-subunit protein by immunoblot relative to untreated monolayers. Fi xed AEC monolayers demonstrated predominantly membrane-associated immu nofluorescent labeling with anti-Na+ pump alpha(1)- and beta(1)-subuni t antibodies, with increased intensity of cell labeling for both subun its seen at 24 h following exposure to EGF. These changes in Na+ pump mRNA and protein preceded a delayed (>12 h) increase in short-current circuit (measure of active transepithelial Naf transport) across monol ayers treated with EGF compared with untreated monolayers. We conclude that EGF increases active Na+ resorption across AEC monolayers primar ily via direct effects on Na+ pump subunit mRNA expression and protein synthesis, leading to increased numbers of functional Na+ pumps in th e basolateral membranes.