The effect of aminoperimidine (AP)on transepithelial Na+ transport and Cl- conductance (G(Cl)) of isolated amphibian skin (Bufo viridis and Rana esculenta) was analyzed using transepithelial and intracellular electrophysiological techniques. AP, applied at concentrations between 30 and 100 mu M from the mucosal side, stimulated Na+ transport rapidly and reversibly by more than 30% of the control value due to an increase in apical membrane Na+ permeability. Influence of AP on basolateral membrane conductance and effective driving force for Na+ were negligible. Voltage-activated G(Cl) of toad skin, but not the resting, deactivated conductance, as well as spontaneously high G(Cl) in frog skin was rapidly inhibited by AP in a concentration-dependent manner. The half-maximal inhibitory concentration of 20 mu M is the highest hithero reported inhibitory power for G(Cl) in amphibian skin. The effect of AP on G(Cl) was slowly and incompletely reversible even after brief exposure to the agent. Serosal application of AP had similar, albeit delayed effects on both Nai and Cl- transport. AP did not interfere with the Cl- pathway after it was opened by 100-300 mu M CPT-cAMP, a membrane-permeable, nonhydrolyzed analogue of cAMP. Inhibition of the voltage-activated G(Cl) by AP was attenuated or missing when AP was applied during voltage perturbation to serosa-positive potentials. Since AP is positively charged at physiological pH, it suggests that the affected site is located inside the Cl- pathway at a certain distance from the external surface. AP affects then the Na+ and Cl- transport pathways independent of each other. The nature of chemical interference with AP, which is responsible for the influence on the transport of Na+ and Cl-, remains to be elucidated.