We have cloned a homolog of Pax-6 in Xenopus laevis. Its deduced amino acid sequence has a 95% overall identity with Pax-6 homologs in other vertebrates. It is expressed early in development in cells fated to form the eye and parts of the forebrain, hindbrain, and spinal cord. It has two phases of expression in the eye. In the early phase, from stage 12.5 to stage 33/34, Xenopus Pax-6 is expressed throughout the developing retina. In the late phase, after stage 33/34, it is excluded from mature cells in the outer half of the retina and from cells in the ciliary marginal zone, remaining only in amacrine and ganglion cells. Misexpression of Pax-6 early in development results in axial defects, but no specific eye phenotype is observed. Targeted misexpression in the retina at later stages does not result in any significant bias toward formation of amacrine or ganglion cells or away from photoreceptors. Ectopic expression of the proneural gene NeuroD alters the pattern of Pax-6, substantially reducing its expression in the eye field and later reducing or eliminating the eye itself. Our results show that Pax-6 expression appears to be necessary, but not sufficient, for eye formation in Xenopus.