The adult mammalian retinal pigment epithelium (RPE) is mitotically inactive, yet retains the capacity to proliferate under certain conditions. To determine the onset of RPE proliferation after retinal detachment, we examined experimentally detached cat retinas of 12, 24, 48, and 72 hr duration. An additional animal served as a nondetached, sham-operated control. 3H-thymidine was injected into the vitreous chamber and the eyes were processed for light microscopic autoradiography. Autoradiograms from both the control and the 12 hr detachment showed no evidence of labeled RPE nuclei; however, labeled nuclei were present at both 24 and 48 hr after detachment. Labeled nuclei per millimeter of RPE at 24 hr were 55% of the 48 hr rate. Mitotic figures were noted only at 48 and 72 hr after detachment. No labeled RPE nuclei appeared in autoradiograms that bordered the detachment zone. Electron micrographs showed that proliferating RPE cells assume several configurations, some of which have been reported in other species. The proliferative response of the RPE occurs much sooner than had previously been thought. It appears to be a local effect that does not involve retinal regions beyond the detachment boundaries, and it may have potentially adverse effects when the retina and RPE are reapposed after retinal reattachment surgery.