Previous studies have demonstrated amiloride-sensitive Na(+) absorption under basal conditions and cystic fibrosis transmembrane conductance regulator (CFTR)-mediated Cl(-) secretion following neurohormonal stimulation in the mouse endometrial epithelium. The present study investigated the inhibition of amiloride-sensitive Na(+) absorption accompanying activation of CFTR in the mouse endometrium using the short-circuit current ( I(sc)) technique. RT-PCR demonstrated the co-expression of CFTR and epithelial Na(+) channels (ENaC) in primary cultured mouse endometrial epithelia and cultured endometrial monolayers exhibited a basal amiloride-sensitive I(sc) of 5.4 +/- 0.6 microA/cm(2). The amiloride-sensitive current fell to 3.1 +/- 0.5 microA/cm(2) after stimulation with forskolin. When the possible contribution of Na(+) absorption to the I(sc) was eliminated by amiloride (1 microM) or Na(+) replacement, the forskolin-induced I(sc) was not reduced, but rather increased significantly compared with that in the absence of amiloride or in Na(+)-containing solutions ( P < 0.02), indicating that the forskolin-induced I(sc) was mediated by Cl(-) secretion, portion of which may be masked by concurrent inhibition of basal Na(+) absorption if the contribution of Na(+) is not eliminated. When the contribution of Cl(-) to the I(sc) was eliminated by diphenylamine 2,2'-dicarboxylic acid (DPC, 2 mM) or Cl(-) replacement, forskolin now decreased, rather than increased the I(sc), demonstrating the inhibition of Na(+) absorption upon stimulation. Our data suggest an interaction between CFTR and ENaC, which may be the underlying mechanism for balancing Na(+) absorption and Cl(-) secretion across the mouse endometrial epithelium.