The normal pattern of innervation of the optic tectum has been studied autoradiographically in a series of chick embryos which were injected intraocularly with [3-H]proline at intervals between the 6th and 21st days of incubation. From the distribution of the radioactively labeled proteins transported in the rapid phase of axonal flow, it is evident that retinal fibers first enter the tectum late on the 6th day of incubation and then spread across its surface from its rostrolateral aspect to its caudo-dorso-medial pole during the ensuing 6 days. At the 9th day, when the fibers have grown across the surface of the rostral half of the tectum, there is no indication that the terminal portions of the axons have left the stratum opticum to enter the outer layers of the stratum grieseum et fibrosum superficiale. The first suggestion of such an invasion of the stratum grieseum et fibrosum superficiale is found at day 10, when labeled fibers can be seen in this stratum, over a restricted, oval, area near the center of the tectum. Over the course of the next 2 days the region of the stratum grieseum et fibrosum superficiale occupied by retinal fibers expands, more-or-less concentrically, until by day 14 some labeled fibers are seen in the outer part of the stratum, throughout the tectum. Concurrent with the ingrowth of optic nerve fibers the final cytoarchitectonic differentiation of the outer layers of the tectum occurs so that by day 18 the autoradiographs show a pattern of labeling of layers a-f of the stratum griseum et fibrosum superficiale similar to that seen in mature, post-hatched chicks. Since the region of the tectum which is first innervated by retinal fibers corresponds to the projection field of the region around the upper end of the choroid fissure where the first ganglion cells are generated, it would appear that the axons of the first-formed ganglion cells grow over the rostral surface of the tectum before establishing the first retino-tectal synapses near the central portion of the tectum. Subsequently, as the ganglion cell population grows concentrically from around the area centralis, there is a parallel expansion of the region in which retino-tectal synapses are being formed in the tectum.