Na+-K+-ATPase activity in medullary thick ascending limb during short-term anoxia

Abstract

Na+-K+-ATPase activity was measured in a suspension of rabbit medullary thick ascending limb tubules under oxygenated and anoxic conditions. Oxygenated, K-depleted tubules rapidly take up added extracellular potassium accompanied by a simultaneous increase in oxygen consumption. The ATP/O2 ratio was 12.5 +/- 0.7, suggesting a tight coupling between oxidative metabolism (6 ATP/O2) and Na+-K+-ATPase activity (2 K/ATP). On reaching anoxia, the tubules released potassium into the medium, but this rate was accelerated by the addition of ouabain, which indicated that the Na+-K+-ATPase was still operative in anoxia. Because 10 min of anoxia led to only a 15.7% decline in potassium content, a new steady state of potassium uptake and leakage must be reached during anoxia. Anaerobic metabolism maintained 73% of cellular ATP during 10 min of anoxia. Exposure of anoxic tubules to iodoacetate produced a 57% decline in ATP levels and a 33% decline in potassium content, which indicated that glycolysis is an important pathway in supplying energy during anaerobiosis.