The ability of homogenates of the chick optic lobe to accumulate a series of possible neurotransmitters has been studied. High affinity uptake of several possible neurotransmitters was examined in optic lobes of 21-day-old embryos that had a single eye removed on the third day of incubation and in 23-day-old chicks that had an eye removed at hatch. Embryonic enucleation resulted in severe reduction of development of the ability of the contralateral optic lobe to take up tritiated GABA, dopamine, choline, serotonin and glutamate from solutions around 10(-8)M. Unilateral eye removal of new-hatched chicks caused failure of the denervated optic lobe to grow, but only the uptake capacity for glutamate was significantly recuced. This deficit was apparent as early as 4 days after enucleation. The transport of other compounds was unimpaired. The uptake of glutamate by homogenates of the optic tract was 43% of that or the optic lobe. This was a much greater fraction than the corresponding value for other postulated neurotransmitters. These data suggest that glutamate may be the primary neurotransmitter of the fibers of the optic tract originating in the retinal ganglion cells.