Application of principles of steady-state kinetics to the estimation of gamma-aminobutyric acid turnover rate in nuclei of rat brain

Abstract

13C-labeled glucose (50 mumol/kg/min) was infused into the rat tail vein for 10 minutes. The animals were killed by a microwave beam focused to the skull at various times after the infusion. In brain nuclei the concentrations of glutamate and gamma-aminobutyric acid (GABA) and their enrichment in 13C were measured by mass fragmentography. The results showed that in the five brain areas studied, substantia nigra contained the highest GABA concentration, followed by globus pallidus, N. accumbens, deep cerebellar nuclei, N. caudatus and cerebellar cortex. The concentrations of glutamate and GABA were comparable in some nuclei (substantia nigra and globus pallidus), but not in others. The glutamate content was 3 to 6 times greater than GABA content in cerebellum (cortex and nuclei) N. accumbens and N. caudatus. The turnover rates of GABA in substantia nigra, globus pallidus, N. caudatus and N. accumbens were calculated by applying principles of steady-state kinetics to the changes with time in the 13C enrichment of glutamic acid and GABA. In substantia nigra, N. accumbens and N. caudatus, the percentage of 13C incorporation into glutamic acid and GABA reached its maximum 10 to 15 minutes after the infusion. In N. caudatus and N. accumbens, the percentage of 13C incorporation into GABA was higher than that in substantia nigra and globus pallidus. The turnover rates of GABA were very similar in N. caudatus, N. accumbens, substantia nigra and globus pallidus (385-562 nmol/mg of protein per hr), but the turnover times were much faster in N. caudatus and N. accumbens. It must be kept in mind that in N. caudatus and N. accumbens the glutamate pool was several-fold greater than that of GABA. Since the data obtained indicate that the compartmentation is more complicated than in the model assumed for turnover estimations, this method should be used only for comparative purposes in drug studies. The validity of this method to obtain absolute measurements of GABA turnover remains to be documented.