This study attempted to determine whether brain serotonin (5-HT), which is altered by melatonin administration, is involved in mediating the effects of melatonin on basal endocrine function. Pineal melatonin levels, serum N-acetylserotonin (NAS) levels, adrenocortical activity, and other endocrine parameters were measured following 5-HT depletion by p-chlorophenylalanine (p-CPA) together with either pineal stimulation by blinding or blinding plus pinealectomy. Blinding increased pineal melatonin levels in both saline and p-CPA treated animals. P-CPA treatment increased adrenal weights and morning plasma corticosterone levels in both blinded and blinded-pinealectomized animals. Conversely, p-CPA depressed pineal melatonin levels and serum NAS but elevated morning plasma corticosterone levels in sighted controls. P-CPA also decreased plasma prolactin and growth hormone levels in intact animals. These findings suggest that 5-HT inhibits morning corticosterone secretion and stimulates prolactin and growth hormone release. In addition, melatonin and serotonin may function independently in regulating adrenocortical function, while melatonin's effect is superceded by that of serotonin.