In male hamsters, exposure to a short photoperiod results in a significant decrease in median eminence (ME) dopamine (DA) concentrations. The mechanism responsible for this decrease in DA is unknown. The experiments described in this paper were designed to examine the effects of photoperiod on DA metabolism and synthesis in the ME to determine if a change in these processes is responsible for the short-photoperiod-induced decrease in ME DA concentrations. In the first experiment, the metabolism of DA in tuberoinfundibular dopamine (TIDA) neuronal terminals was determined by measuring ME concentrations of 3,4-dihydroxyphenylacetic acid (DOPAC; a major metabolite of DA) and DA in male and female hamsters housed in long and short photoperiods. In both males and females, exposure to the short photoperiod induced a collapse of the reproductive system and a reduction in circulating prolactin. In males, but not in females, exposure to the short photoperiod reduced ME DA concentrations; however, DOPAC concentrations were not affected by photoperiod. Thus, the decrease in ME DA seen in males is not the result of an increase in DA metabolism. In the second experiment, tyrosine hydroxylase (TH) activity in the ME of males was determined by injecting animals housed in long and short photoperiods with a L-aromatic amino acid decarboxylase inhibitor (NSD 1015) and measuring 3,4-dihydroxyphenylalanine (DOPA). Consistent with Experiment 1, ME DA concentrations were significantly decreased in gonadally regressed males housed in a short photoperiod; however, ME DOPA accumulation was not affected. Thus, the observed decrease in DA is not the result of a decrease in TH activity in the ME. The results of the experiments presented here indicate that (1) in males but not females, the decrease in circulating prolactin seen in animals housed in a short photoperiod for 12 weeks is associated with a decrease in ME DA concentrations, and (2) the decrease in ME DA seen in males housed in a short photoperiod is not the result of an increase in DA metabolism or a decrease in synthesis by TIDA neurons.