Basis for apparent saturation kinetics of Na+ influx in freshwater hyperregulators

Abstract

Uptake of Na+ by intact frogs has been reported to show saturation kinetics at low external concentrations (less than 2 mM); yet other evidence shows that the transport system is far from saturated in this concentration range. The saturation behavior was reproduced here in isolated frog skins that were then used as appropriate models for investigating the paradox. When the skin was bathed by 2 mM Na+ outside and open circuited, influx (JNain) was near maximum. If, under these conditions, the skin was short circuited, JNain increased threefold. Alternatively, if Cl- permeability was increased in the open-circuited skin, JNain doubled. Both perturbations uncouple JNain from the efflux of a cation (nominally H+), which normally maintains electrical neutrality under open-circuit conditions. This suggests that the apparent saturation of JNain is caused by limiting efflux of the counterion. In confirmation of this prediction, stimulation of proton efflux markedly increased JNain. Thus the apparent Michaelis-Menten kinetics observed in frogs, and probably in other freshwater animals as well, do not represent saturation of an element in Na+ transport, either the amiloride-sensitive apical channel or the basolateral Na+-K+-ATPase.