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ENaC subunit mRNA in the collecting duct cell line by a p38 MAPK-dependent pathway
1 Graduate Program in Molecular Biology, University of Iowa College of Medicine, Iowa City, IA, USA
2 Department of Internal Medicine, University of Iowa College of Medicine, Iowa City, IA, USA
3 Department of Internal Medicine, University of Iowa College of Medicine, Iowa City, IA, USA; Graduate Program in Molecular Biology, University of Iowa College of Medicine, Iowa City, IA, USA; Veterans Affairs Medical Center, Iowa City, IA, USA
* To whom correspondence should be addressed. E-mail: christie-thomas{at}uiowa.edu.
Aldosterone and glucocorticoids (GC) stimulate Na+ reabsorption in the collecting ducts by increasing the activity of ENaC. We have used MDCK-C7 cells to demonstrate that this effect is associated with an increase in 
ENaC gene transcription (32). Cycloheximide (CHX), superinduced the GC-stimulated ENaC expression in a dose-dependent manner, but had no effect on basal or aldosterone-stimulated ENaC expression, while anisomycin inhibited basal and corticosteroid-stimulated ENaC expression. The superinduction of ENaC expression was also seen with hypotonicity, was blocked by RU38486, and was independent of protein synthesis. CHX had no effect on ENaC mRNA half-life confirming that its effect was via an increase in ENaC transcription. The effect of CHX and hypotonicity on ENaC expression was abolished by SB202190, indicating an effect mediated via p38 MAPK. Consistent with this scheme CHX increased pp38 and MKK6, an upstream activator of p38, stimulated ENaC promoter activity. These data confirm a model where CHX activates p38 in MDCK-C7 cells to increase ENaC gene transcription in a GC-dependent manner.
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