Regulation of Na+/K+-ATPase activity by nitric oxide in the kidney and gill of the brown trout (Salmo trutta).
In teleost fish, successful osmoregulation involves controlled ion transport mechanisms in kidney and gill epithelia. In this study, the effect of nitric oxide (NO) on Na(+)/K(+)-ATPase was investigated in vitro in these two tissues in brown trout (Salmo trutta) acclimated to freshwater. The NO donor sodium nitroprusside (SNP) inhibited in situ Na(+)/K(+)-ATPase activity, measured as ouabain-sensitive Rb(+) uptake, in both samples of kidney and gill tissue and in isolated gill cells. The effect was dose-dependent in both tissues, with a maximal observed inhibition of approximately 40-50% (1 mmol l (-1) SNP). The time-course of inhibition revealed a maximum effect with 10 min pre-incubation. The effect of SNP was reproduced with another NO donor, papa-nonoate (NOC-15; 200 micro mol l(-1)), and was prevented by the NO scavenger 2-phenyl-4,4,5,5-tetramethylimidazoline-1-oxyl 3-oxide (PTIO; 1 mmol l(-1)). To further investigate the mechanism of the NO effect, whole-tissue Na(+) and K(+) levels were analysed. In kidney, SNP (1 mmol l(-1)) led to an increase in tissue Na(+) levels and a decrease in K(+) levels in a 3:2 ratio. In gill tissue, no change in either ion was observed. These observations indicate that the effect on Na(+)/K(+)-ATPase is direct rather than due to a decrease in intracellular Na(+), its rate-limiting substrate. SNP elevated the level of cyclic GMP (cGMP) in both kidney and gill tissue. Dibutyryl cyclic GMP (db-cGMP; 1 mmol l(-1)) also inhibited Na(+)/K(+)-ATPase activity in both tissues. Hence, a possible mechanism may involve the cGMP-activated kinase, even though other mechanisms cannot be excluded.
Tipsmark
Christian K
CK
Institute of Biology, University of Southern Denmark, Odense University, Campusvej 55, DK-5230 Odense M, Denmark.
Madsen
Steffen S
SS
D000704:Analysis of Variance; D000818:Animals; D006152:Cyclic GMP / Q000494:pharmacology; D005880:Gills / Q000187:drug effects*; D066298:In Vitro Techniques; D007668:Kidney / Q000187:drug effects*; D009569:Nitric Oxide / Q000494:pharmacology*; D009599:Nitroprusside / Q000494:pharmacology; D000254:Sodium-Potassium-Exchanging ATPase / Q000037:antagonists & inhibitors*; D014337:Trout / Q000378:metabolism*; D014882:Water-Electrolyte Balance / Q000187:drug effects
D009599:Nitroprusside; D009569:Nitric Oxide; D000254:Sodium-Potassium-Exchanging ATPase; D006152:Cyclic GMP