Quantification of the GABA Shunt and the Importance of the GABA Shunt Versus the 2‐Oxoglutarate Dehydrogenase Pathway in GABAergic Neurons

Abstract: We investigated the activity of the cerebral GABA shunt relative to the overall cerebral tricarboxylic acid (TCA) cycle and the importance of the GABA shunt versus 2‐oxoglutarate dehydrogenase for the conversion of 2‐oxoglutarate into succinate in GABAergic neurons. Awake mice were dosed with [1‐ 13 C]glucose, and brain extracts were analyzed by 13 C NMR spectroscopy. The percent enrichments of GABA C‐2 and glutamate C‐4 were the same: 5.0 ± 1.6 and 5.1 ± 0.2%, respectively (mean ± SD). This, together with previous data, indicates that the flux through the GABA shunt relative to the overall cerebral TCA cycle flux equals the GABA/glutamate pool size ratio, which in the mouse is 17%. It has previously been shown that under the experimental conditions used in this study, the 13 C labeling of aspartate from [1‐ 13 C]glucose specifically reflects the metabolic activity of GABAergic neurons. In the present study, the reduction in the formation of [ 13 C]aspartate during inhibition of the GABA shunt by γ‐vinyl‐GABA indicated that not more than half the flux from 2‐oxoglutarate to succinate in GABAergic neurons goes via the GABA shunt. Therefore, because fluxes through the GABA shunt and 2‐oxoglutarate dehydrogenase in GABAergic neurons are approximately the same, the TCA cycle activity of GABAergic neurons could account for one‐third of the overall cerebral TCA cycle activity in the mouse. Treatment with γ‐vinyl‐GABA, which increased GABA levels dramatically, caused changes in the 13 C labeling of glutamate and glutamine, which indicated a reduction in the transfer of glutamate from neurons to glia, implying reduced glutamatergic neurotransmission. In the most severely affected animals these alterations were associated with convulsions.