UPTAKE OF [BENZAMIL-H-3 AT DIFFERENT SODIUM CONCENTRATION - INFERENCES REGARDING THE REGULATION OF SODIUM PERMEABILITY

1. The effect of benzamil on short‐circuit current in frog skin was measured at different external sodium concentrations. A linear relationship exists between the concentration of benzamil reducing short‐circuit current by 50% and the external sodium concentration, indicative of some form of competitive antagonism between sodium and benzamil. 2. Uptake of [3H]benzamil into isolated frog skin epithelium and whole skin (0.95 cm2 pieces) was measured at different external sodium concentrations. With a sodium concentration of 111 mM in the external medium the uptake of [3H]benzamil is linear with concentration. Uptake amounted to 8.8 f‐mole nM‐1, a value similar to the linear component of the uptake measured at low (1.1 mM) sodium concentration. 3. Using a variety of other conditions the maximal number of specific binding sites for [3H]benzamil was calculated from displaceable binding and the fractional occupancy, the latter being derived from the inhibition of short‐circuit current. This approach gave similar binding site densities to those reported previously at low sodium concentrations. 4. The reduction in specific [3H]benzamil uptake at high sodium may result from two mechanisms, competition of sodium with the ligand for an external binding site and a reduction in the site density as the intracellular sodium concentration increases. 5. It is concluded that the saturation of sodium transport which occurs at high sodium concentration is likely a consequence of the reduced availability of entry sites, rather than saturation of the uptake process. © 1979 The Physiological Society