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The effect of tryptophan administration to pregnant rats on the development of serotonergic systems and serotonin-related hormones in the offspring was studied. The male offspring of rats treated with tryptophan (200 mg/kg/day) during the second half of gestation showed a 4- to 7-fold increase in serum prolactin 40 and 70 days after birth and a 2-fold increase in serum luteinizing hormone 70 days after birth. The forebrain of adult offspring of tryptophan-treated rats showed an increase in serotonin and 5-hydroxyindoleacetic acid levels. Present data suggest that tryptophan regulates serotonergic differentiation during early development.