GABA alters the metabolic fate of [U-13C]glutamate in cultured cortical astrocytes

The effect of gamma-aminobutyric acid (GABA) on glutamate metabolism was studied by (13)C-nuclear magnetic resonance (NMR) spectroscopy. Cerebral cortical astrocytes were incubated with 0.5 mM [U-(13)C]glutamate and 5 mM glucose in the presence or absence of 0.2 mM GABA for 2 hr. (13)C-labeled glutamate, glutamine, and aspartate were observed in cell extracts, and (13)C-labeled glutamine and lactate were present in the media. Both uniformly labeled glutamate and [1,2,3-(13)C]glutamate derived from the tricarboxylic acid (TCA) cycle were present in the cells. The consumption of [U-(13)C]glutamate and glucose was unchanged in the presence of GABA; however, the formation of [U-(13)C]lactate and [U-(13)C]aspartate from metabolism of [U-(13)C]glutamate was increased in cells incubated with GABA. The total concentration of aspartate was increased to the same extent as the (13)C-labeled aspartate, suggesting increased entry of [U-(13)C]glutamate into the TCA cycle to allow for the transamination of GABA. Although the concentrations of unlabeled glucose and lactate in the media were unchanged in the presence of GABA, the concentration of alanine was decreased, indicating that there was decreased transamination of the unlabeled pyruvate from glucose metabolism. The amount of [U-(13)C]glutamate converted to [U-(13)C]glutamine and [U-(13)C]lactate was increased in the presence of GABA. However, since the overall consumption of [U-(13)C]glutamate was not different, it can be concluded that the amount of [U-(13)C]glutamate used for energy was decreased. This suggests that exogenous GABA could substitute for glutamate as an energy source for astrocytes. The results indicate that the presence of GABA influences the metabolic fate of both glutamate and glucose in astrocytes, suggesting that fluctuations in the concentration of GABA in normal and pathological conditions can alter the compartmentation of glial metabolism in brain.