l-Proline Alters Energy Metabolism in Brain Cortical Tissue Slices

L-Proline (l-Pro) is a non-essential amino acid which, in high concentrations, can cause neurological problems including seizures, although the causative mechanism for this is unclear. Here, we studied the impact of physiological levels of proline on brain energy metabolism and investigated the metabolism of l-Pro itself, using the cortical brain tissue slice and stable isotope labelling from [1-¹³ C]glucose and [1,2-¹³ C]acetate detected by NMR spectroscopy and LCMS. l-Pro was actively taken up by the slices and significantly reduced the total metabolic pools of all measured metabolites with glutamine the least affected, while reducing net flux of ¹³C into glycolytic byproducts (lactate and alanine). Conversely, net flux into Krebs cycle intermediates was increased, suggesting that L-Pro at lower concentrations was driving increased mitochondrial activity in both neurons and glia at the expense of glycolysis and metabolic pool sizes. As there was no evidence of metabolism of [1-¹³ C] l-Pro in slices under normo-glycemic conditions, the effect of proline on metabolism was not due to displacement of metabolites by added l-Pro. Comparison of the metabolic fingerprint generated by l-Pro in slices metabolizing [3-¹³ C]pyruvate with that generated by ligands active in the GABAergic system suggested that l-Pro may engender effects similar to that of the inhibitory neurotransmitter and metabolite γ-aminobutyric acid (GABA), in line with previous suggestions that l-Pro may be a GABA mimetic in addition to its role as a modulator of mitochondrial metabolism.