To compare the roles of the nonreceptor tyrosine kinase c-src and its neuronal splice form n-src in developing neurons, Xenopus retinal precursors were transfected in vivo with c-src, n-src, or constitutively active mutants. Axonogenesis of retinal ganglion cells was markedly impaired by the expression of constitutively active c-src and only mildly affected by the expression of constitutively active n-src. This differential phenotype could not be accounted for by raised levels of intracellular tyrosine phosphorylation alone because the average anti-phosphotyrosine staining intensity of retinal neurons expressing mutant n-src was almost twofold greater than that of neurons expressing mutant c-src. The expression of either constitutively active isoform inhibited photoreceptor differentiation by 72% but did not influence other cell fates. These results suggest that c-src and n-src have both overlapping and distinct activities in differentiating retinal neurons.