Vanadate and ouabain: a comparative study in toad skin.

In this study we compare the effects of two inhibitors of the Na,K-ATPase, ouabain and vanadate, upon transport properties of the isolated short-circuited toad skin: The main conclusions are: Both inhibitors induce a similar decline in short-circuit current (SCC). They differ regarding skin electrical resistance (R). Ouabain induces an increase in resistance that, after some delay, builds up slowly after its addition to the preparation, while vanadate causes a fast increase in resistance that remains constant for most of the experimental period. Vanadate, but not ouabain, promotes an unspecific increase in skin permeability characterized by a delayed and progressive rise of 42K (JK eff) and 14C sucrose (J suc eff) effluxes. Vanadate effect upon skin permeability, as measured by JK eff, is not affected by pre-treating the skin with DIDS, a stilbene derivative, indicating that anion-exchange is not an important step for the entrance of vanadate into the epithelial cells to trigger its effect. Vanadate effect upon JK eff is also not affected by previous ouabain inhibition of the Na,K-ATPase, showing that this effect is not mediated by the inhibition of this enzyme. Vanadate action in toad skin seems to occur at junctional structures opening paracellular routes. A possible mechanism for the effect of vanadate is discussed in terms of cytosolic Ca2+ balance, cytoskeleton and their interplay with the sealing of tight junctions.