This study investigated the transport rate of the tracer, cholera toxin B subunit, within the retinofugal pathway of the chick hatchlings. Following intraocular injections, the chicks were allowed to survive for various time-periods. The immunoreactivity of cholera toxin B subunit was then examined in the retinofugal pathways. Two hours post-injection, retinal ganglion cells began to take up the tracer and transport it to the most rostroventral portion of the optic tectum. After a 4 h survival period, the labeled retinal axons progressively innervated all retinofugal targets. Within the tectum, the labeling density varied from layer to layer with heavily labeled terminals in layer 5b, less label in layer 7 and the most diffuse label in layers 2-4. Scattered labeling was seen in the nucleus dorsolateralis anterior thalami, pars lateralis, the nucleus geniculatus lateralis, pars ventralis, the nucleus basal optic root, the nucleus lateralis anterior thalami, and the pretecal lentiformis nucleus of mesencephalon. After 6- and 8 h survival periods, increased labeling was seen in all retinofugal nuclei. There were increased numbers of retinal terminals in all retinorecipient layers of the tectum. It was noted that some of the retinal axons "overshot" into layers deeper than layer 7. In addition, retinal projections were found scattered throughout the ipsilateral nucleus basal optic root. Maximal labeling in all retinofugal targets was observed at a 10 h survival period. The present study suggests that cholera toxin B subunit can be used to trace retinal axons along their retinofugal paths up to the small terminal zones at a rate of 4.25 mm/h or 102 mm/day. Also, evidence of synchronous retinal terminations in layers 5b and 7 indicates that the transport of cholera toxin B subunit is independent of axon diameters of retinal ganglion cells. Finally, given the changing status of the embryo, the rapid transport of cholera toxin B subunit can be applied for tracing developing pathways.