We have reported previously that during optic nerve regeneration in Rana pipiens, axons are misrouted into the opposite nerve and retina. In the present investigation we have examined the time course of formation of these "misrouted" axons and their cells of origin. The right eye of 31 frogs was injected with 3H-proline at various times after right optic nerve crush. In every frog examined 2 weeks and later after nerve crush, the distribution of autoradiographic label indicated that axons from the right eye had grown into the left optic nerve at the chiasm. The amount of label increased from 2 weeks to reach a maximum at 6 weeks where the entire left nerve was filled with silver grains. At 5 to 6 weeks after crush, labeled axons were found within the ganglion cell fiber layer (GCFL) of the retina of the opposite eye for a maximum distance of 2.3 mm from the optic disc. In frogs examined at intervals later than 6 weeks after crush, the amount of label within the left eye and nerve progressively decreased, indicating a gradual disappearance of the misrouted axons. Studies using anterograde transport of horseradish peroxidase (HRP) after nerve injection confirmed these autoradiographic findings. The position of ganglion cells in the right eye whose axons were misrouted to the left eye was determined by retrograde transport of HRP. Five or 6 weeks after crushing the right optic nerve, the left eye was injected with HRP and labeled ganglion cells were found throughout the right eye retina. The largest percentage of labeled cells was found within the ventral half of the retina, particularly within the temporal quadrant, and nearly all of the labeled cells were found in more peripheral portions of the retina. Since few retino-retinal axons are found during normal development, the present results show that the factors guiding regenerating axons in the adult frog differ substantially from those present during development.