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Am J Physiol Lung Cell Mol Physiol 281: L217-L230, 2001;
1040-0605/01 $5.00
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Vol. 281, Issue 1, L217-L230, July 2001

Modulation of alpha -ENaC and alpha 1-Na+-K+-ATPase by cAMP and dexamethasone in alveolar epithelial cells

André Dagenais1, Christine Denis1, Marie-France Vives1, Sonia Girouard1, Chantal Massé1, Thao Nguyen1, Toshiyuki Yamagata1, Czeslawa Grygorczyk1, Rashmi Kothary2, and Yves Berthiaume1

1 Département de Médecine, Centre de Recherche, Centre Hospitalier de l'Université de Montréal-Hôtel-Dieu, Université de Montréal, Montreal, Quebec H2W 1T8; and 2 Centre for Molecular Medicine, Ottawa General Hospital Research Institute, Ottawa, Ontario K1H 8L5, Canada

cAMP and dexamethasone are known to modulate Na+ transport in epithelial cells. We investigated whether dibutyryl cAMP (DBcAMP) and dexamethasone modulate the mRNA expression of two key elements of the Na+ transport system in isolated rat alveolar epithelial cells: alpha -, beta -, and gamma -subunits of the epithelial Na+ channel (ENaC) and the alpha 1- and beta 1-subunits of Na+-K+-ATPase. The cells were treated for up to 48 h with DBcAMP or dexamethasone to assess their long-term impact on the steady-state level of ENaC and Na+-K+-ATPase mRNA. DBcAMP induced a twofold transient increase of alpha -ENaC and alpha 1-Na+-K+-ATPase mRNA that peaked after 8 h of treatment. It also upregulated beta - and gamma -ENaC mRNA but not beta 1-Na+-K+-ATPase mRNA. Dexamethasone augmented alpha -ENaC mRNA expression 4.4-fold in cells treated for 24 h and also upregulated beta - and gamma -ENaC mRNA. There was a 1.6-fold increase at 8 h of beta 1-Na+-K+-ATPase mRNA but no significant modulation of alpha 1-Na+-K+-ATPase mRNA expression. Because DBcAMP and dexamethasone did not increase the stability of alpha -ENaC mRNA, we cloned 3.2 kb of the 5' sequences flanking the mouse alpha -ENaC gene to study the impact of DBcAMP and dexamethasone on alpha -ENaC promoter activity. The promoter was able to drive basal expression of the chloramphenicol acetyltransferase (CAT) reporter gene in A549 cells. Dexamethasone increased the activity of the promoter by a factor of 5.9. To complete the study, the physiological effects of DBcAMP and dexamethasone were investigated by measuring transepithelial current in treated and control cells. DBcAMP and dexamethasone modulated transepithelial current with a time course reminiscent of the profile observed for alpha -ENaC mRNA expression. DBcAMP had a greater impact on transepithelial current (2.5-fold increase at 8 h) than dexamethasone (1.8-fold increase at 24 h). These results suggest that modulation of alpha -ENaC and Na+-K+-ATPase gene expression is one of the mechanisms that regulates Na+ transport in alveolar epithelial cells.

sodium channel; adenosine 3',5'-cyclic monophosphate; steroid; sodium-potassium-adenosinetriphosphatase; transepithelial current; epithelial sodium channel


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