The roles of epithelial sodium channel (ENaC) subunits (, , and ) in the impaired renal reabsorption of sodium and water were examined in rat models with bilateral, or unilateral ureteral obstruction (BUO and UUO) for 24 hours and BUO followed by release of obstruction and observed for 3 days (BUO-3dR). In BUO rats, plasma osmolality was increased dramatically whereas plasma sodium concentration was decreased. Immunoblotting revealed a significant decreased expression of -ENaC (57 ± 7%), -ENaC (19 ± 5%), and -ENaC (51 ± 10%) as well as 11-hydroxysteroid dehydrogenase type 2 (11-HSD2) in the cortex and outer medulla (C+OM) compared with sham-operated controls. This was confirmed by immunohistochemistry. BUO-3dR was associated with polyuria and impaired renal sodium handling. The protein abundance and the apical labeling of -ENaC were significantly increased whereas - and -ENaC as well as 11-HSD2 expression remained decreased. In UUO rats, expression of -, -ENaC and 11-HSD2 decreased in the C+OM in the obstructed kidney. In contrast, the abundance and the apical labeling of -ENaC in the non-obstructed kidneys were markedly increased, suggesting compensatory upregulation in this kidney. In conclusion, -, -, and -ENaC expression levels are downregulated in the obstructed kidney. The expression and apical labeling of -ENaC were increased in BUO-3dR rats and in the non-obstructed kidneys from UUO rats. These results suggest that altered expression -, -, and -ENaC may contribute to impaired renal sodium and water handling in response to ureteral obstruction.