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This study observed the effect of giving tryptophan on the levels of Luteinizing Hormone (LH) and the number of Leydig cells. The study used the design of the post test only control group design, where the sample was dividedinto 4 groups consisting of 1 control group without treatment and 3 treatment groups with the provision of tryptophan doses of 40mg, 50mg and 60mg respectively. The treatments are given for 14 days and measured hormone levels using RIA. Counting the number of Leydig cells contained in the interstitial network between seminiferous tubules using an electric microscope. The results showed a statistically significant effect, there was a decline in the average levels of the hormone LH from 5.60 ± 0,30 nmol/L and a decreased in the number of leydig cells by 20.20 ± 2.06%. The levels of Luteinizing Hormone and the number of Leydig cells decreased in accordance with increasing the dose of tryptophan given on the white male rats Rattus Novergicus when compared with the control group. It could be concluded that the tryptophan that works as a neurotransmitter/ serotonin is able to affect the hypothalamus which then to the anterior pituitary which works as a regulator of the secretion of reproductive hormones.