Antunes, B. C.; Faria-Pereira, A.; Goncalves Ribeiro, J.; Costa Pinto, S.; Vaz, S. H. A.; Morais, V. A.

Mitochondria process glucose, glutamine, and fatty acids (FAs) to produce ATP, with fuel choice dependent on tissue-specific metabolism. The brain harbors two distinct mitochondrial populations - synaptic and non-synaptic. While glucose is the primary fuel for brain bioenergetics, the role of FAs remains elusive. A preliminary proteomic analysis revealed that synaptic mitochondria favor FA metabolism, corroborated by biochemical and respiratory assays showing their higher capacity for {beta}-oxidation and greater respiratory flexibility compared to non-synaptic mitochondria. Additionally, synaptic mitochondria showed higher capacity for FA uptake and less susceptibility to inhibition of the carnitine shuttle system. In neurons, oxygen consumption rate assays indicate that medium to long-chain FA fueling enhances neuronal respiratory flexibility and ATP content, while whole-cell patch-clamp recordings show that long-chain FA fueling sustains increased pre-synaptic activity. Our findings demonstrate that FAs can contribute effectively to synaptic metabolism under normal physiological conditions, where there is a constant demand for energy.