Metabolic reprogramming during neuronal differentiation

Top of pageAbstract Newly generated neurons pass through a series of well-defined developmental stages, which allow them to integrate into existing neuronal circuits. After exit from the cell cycle, postmitotic neurons undergo neuronal migration, axonal elongation, axon pruning, dendrite morphogenesis and synaptic maturation and plasticity. Lack of a global metabolic analysis during early cortical neuronal development led us to explore the role of cellular metabolism and mitochondrial biology during ex vivo differentiation of primary cortical neurons. Unexpectedly, we observed a huge increase in mitochondrial biogenesis. Changes in mitochondrial mass, morphology and function were correlated with the upregulation of the master regulators of mitochondrial biogenesis, TFAM and PGC-1α. Concomitant with mitochondrial biogenesis, we observed an increase in glucose metabolism during neuronal differentiation, which was linked to an increase in glucose uptake and enhanced GLUT3 mRNA expression and platelet isoform of phosphofructokinase 1 (PFKp) protein expression. In addition, glutamate–glutamine metabolism was also increased during the differentiation of cortical neurons. We identified PI3K–Akt–mTOR signalling as a critical regulator role of energy metabolism in neurons. Selective pharmacological inhibition of these metabolic pathways indicate existence of metabolic checkpoint that need to be satisfied in order to allow neuronal differentiation. Abbreviations: MS, mass spectrometry; LC, liquid chromatography; DIV, day in vitro; DON, 6-diazo-5-oxo-l-norleucine; 2-DG, 2-deoxy-d-glucose; CAF, chloramphenicol; MEF-2, myocyte enhancer factor-2; mTOR, mammalian target of rapamycin; FCCP, carbonyl cyanide 4-(trifluoromethoxy) phenylhydrazone; GSH, glutathione; GSSG, glutathione disulphide; NAC, N-acetylcysteine; GCLM, glutamate-cysteine ligase modifier subunit; Gpx1, glutathione peroxidase 1; PFKp, platelet isoform of phosphofructokinase 1; Sod1, superoxide dismutase 1; Cox2, cyclooxygenase 2; tfb2m, transcription factor B2, mitochondrial