Dihydroouabain, a reversible inhibitor of the sodium pump in frog skin

Abstract

In many studies of the sodium pump in epithelia, a readily reversible analog of ouabain would be most useful. This would enable studies of pump activity to be made under control and experimental conditions on the same tissue. Of three compounds examined on the basolateral membrane of the isolated epithelia of frog skin, dihydroouabain (DHO) had characteristics very similar to ouabain except that it was apparently much more reversible. DHO (1 mmol/l) inhibited short circuit current (Isc) and transepithelial Na flux (JNa13) in a fashion similar to ouabain. Isc was inhibited from 17.0 +/- 2.5 to 10.2 +/- 1.0 microA/cm2 in 2-4 min while JNa13 was decreased from 16.8 +/- 1.9 to 4.7 +/- 0.8 microA/cm2 in the same time interval. After 60 min of washout, Isc and JNa13 recovered to about 70% of control values and were nearly equal. In another set of experiments, the washout of DHO and ouabain were compared directly on the same tissue. Sodium flux recovered four times faster after removal of DHO when compared to ouabain. Pretreatment of tissues with DHO prior to ouabain greatly increased the rate of Na flux recovery after washout of both drugs suggesting that DHO competes for ouabain sites. These data suggest that DHO can be used as a reversible analog for ouabain in studies of the Na pump in frog skin.