Understanding the bioenergetics of axon extension and maintenance has wide ranging implications for neurodevelopment and disease states. Glycolysis is a pathway consisting of ten enzymes and separated into preparatory and payoff phases, the latter producing ATP. Using embryonic chicken sensory neurons, we report that glycolytic enzymes are found through the axon and the growth cone. Pharmacological inhibition of glycolysis in the presence of NGF impairs axon extension and growth cone dynamics within minutes without affecting axon maintenance. Experiments using microfluidic chambers show the effect of inhibiting glycolysis on axon extension is local along distal axons and can be reversed by promoting mitochondrial respiration. Knock down of GAPDH simplifies growth cone morphology and is rescued by shRNA resistant GAPDH expression. Rescue of GAPDH using killer red (KR) fused to GAPDH followed by localized chromophore assisted light inactivation of KR-GAPDH in distal axons halts growth cone dynamics. Consider...