The nature of the linkage between between branchial ammonia excretion (JAmm) and unidirectional Na+ influx (JNain) was studied in the freshwater rainbow trout (Oncorhynchus mykiss). Arterial plasma total [ammonia], PNH3 and JAmm were all elevated approximately threefold by intravascular infusion for 24 h with either 70 mmol l-1 (NH4)2SO4 or 140 mmol l-1 NH4HCO3 at a rate of approximately 400 micromol kg-1 h-1. Both treatments markedly stimulated JNain. NH4HCO3 induced metabolic alkalosis in the blood plasma, whereas (NH4)2SO4 caused a slight metabolic acidosis. Experiments with Hepes-buffered water (5 mmol l-1) under control conditions demonstrated that increases in gill boundary layer pH were associated with decreases in both JNain and JAmm. Thus, the stimulation of JNain caused by ammonium loading was not simply a consequence of a Na+-coupled H+ extrusion mechanism activated by internal acidosis or by alkalosis in the gill boundary layer. Indeed, there was no stimulation of net acidic equivalent excretion accompanying NH4HCO3 infusion. Michaelis-Menten kinetic analysis by acute variation of water [Na+] demonstrated that both infusions caused an almost twofold increase in JNamax but no significant change in Km, indicative of an increase in transporter number or internal counterion availability without an alteration in transporter affinity for external Na+. The increase in JNain was larger with (NH4)2SO4 than with NH4HCO3 infusion and in both cases lower than the increase in JAmm. Additional evidence of quantitative uncoupling was seen in the kinetics experiments, in which acute changes in JNain of up to threefold had negligible effects on JAmm under either control or ammonium-loaded conditions. In vitro measurements of branchial Na+/K+-ATPase activity demonstrated no effect of NH4+ concentration over the concentration range observed in vivo in infused fish. Overall, these results are consistent with a dominant role for NH3 diffusion as the normal mechanism of ammonia excretion, but indicate that ammonium loading directly stimulates JNain, perhaps by activation of a non-obligatory Na+/NH4+ exchange rather than by an indirect effect (e.g. Na+-coupled H+ excretion) mediated by altered internal or external acid-base status.
The linkage between Na+ uptake and ammonia excretion in rainbow trout: kinetic analysis, the effects of (NH4)2SO4 and NH4HCO3 infusion and the influence of gill boundary layer pH
- Views Icon Views
- Share Icon Share
- Search Site
A. Salama, I.J. Morgan, C.M. Wood; The linkage between Na+ uptake and ammonia excretion in rainbow trout: kinetic analysis, the effects of (NH4)2SO4 and NH4HCO3 infusion and the influence of gill boundary layer pH. J Exp Biol 15 March 1999; 202 (6): 697–709. doi: https://doi.org/10.1242/jeb.202.6.697
Download citation file: