Uptake of 3H-serotonin is localized to the outer plexiform layer in Long-Evans rat retinas. Autoradiographic accumulation is seen only after in vitro incubation in the light, with retinas isolated from the underlying sclera. Potassium stimulates the release of 3H-serotonin. In this species, amacrine cells do not accumulate these compounds; thus the outer plexiform layer appears to be the only site of uptake and release of this indoleamine. The age-related loss of 3H-serotonin accumulation in the outer plexiform layer of retinal dystrophic rats coincides temporally with the spontaneous degeneration of photoreceptor cells that occurs in this species. Electron-microscopic autoradiography of 3H-serotonin accumulation further confirms that uptake is localized to rod and cone terminals in the outer plexiform layer. The specific accumulation of indoleamines into rod and cone terminals that is observed in the light but is absent in darkness suggests that indoles have an important physiological role in photoreceptors.