Effects of anions on the Na(+)-H+ exchange of trout red blood cells

1. Replacement of chloride by foreign anions in the suspending medium of trout erythrocytes can affect in a complex manner both the activation by catecholamines of the latent Na(+)-H+ exchanger and its subsequent desensitization. These changes are discussed in relation to other cellular modifications (distribution of permeant anions and accumulation of cyclic AMP) induced by foreign anions. 2. The transfer of trout erythrocytes from a chloride-containing medium to media containing lyophilic permeable anions, NO3- or SCN-, immediately induces a decrease of distribution ratios of permeable anions across the red cell membrane (i.e. Donnan ratios). It is probable that the binding of lyophilic anions to haemoglobin, by altering the amount of negative fixed charges, results in changes of distribution of permeant anions across the membrane. 3. The effectiveness of anions in decreasing both the activation of the Na(+)-H+ exchanger and the Donnan ratio follows the same sequence in both cases, i.e., SCN- greater than NO3- greater than Cl- = propionate. It was demonstrated that a change in Donnan ratio affects antiport activity possibly through a shift in intracellular pH; such a mechanism however cannot account for all the effects of foreign anions on antiport activity. 4. The present results show that lyophilic anions do not modify the affinity of the antiporter for sodium ions but greatly decrease the transport capacity of the exchange system. This is interpreted as indicating that the binding of lyophilic anions to some component of the transport system prevents antiporters from establishing their activated configuration once stimulated. Since the inhibitory effect of anions on Na(+)-H+ exchange has been demonstrated in all erythrocytes studied but in no other cell, the crucial substance involved in this inhibition could well be haemoglobin, which appears to control antiport activity in erythrocytes. 5. Some anions affect desensitization of the exchanger. This effect is not related to the lyophilic character of the anion and is not mediated by a change in intracellular cyclic AMP. 6. Propionate and acetate drastically reduce the intracellular level of cyclic AMP and seem to facilitate the activated configuration of the exchanger.