Interference with cellular energy metabolism reduces kynurenic acid formation in rat brain slices: reversal by lactate and pyruvate.

Abstract

This study was designed to investigate the role of cellular energy metabolism in the de novo formation of the endogenous excitatory amino acid receptor antagonist, kynurenic acid. Using rat cortical tissue slices, the roles of glucose transport, glycolysis, tricarboxylic acid cycle intermediates and oxidative phosphorylation were studied. Inhibition of glucose utilization resulted in quantitatively similar decreases in kynurenine uptake, kynurenic acid production and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) reduction, a marker of mitochondrial activity. The end product of glycolysis, pyruvate, as well as lactate, attenuated all three deficits. Pyruvate also significantly increased kynurenic acid formation in normal brain slices without affecting kynurenine uptake. Oxaloacetate and alpha-ketoglutarate (tricarboxylic acid cycle intermediates) were the only compounds tested which were capable of duplicating the effects of pyruvate, indicating that 2-oxoacids can stimulate kynurenic acid synthesis by acting as aminoacceptors in the enzymatic transamination of kynurenine. When the mitochondrial electron transport chain was blocked by specific inhibitors, coincubation with succinate restored the rate of MTT formazan formation to normal (except in the case of 3-nitropropionic acid), yet failed to prevent the resulting reduction in kynurenic acid synthesis. Conversely, pyruvate increased kynurenic acid production in the presence of all inhibitors (except cyanide), but did not attenuate the reduction in kynurenine uptake and MTT formazan formation. Taken together, these results demonstrate that interference with cellular energy metabolism causes mechanistically diverse, pronounced reductions in the cerebral neosynthesis of kynurenic acid, and that 2-oxoacids and lactate can effectively reverse most of these detrimental effects.