In seasonal breeders like the hamster, gonadal function depends on the photoperiod. The present study examined the effects of different photoperiods and ambient temperatures on the gonadotropin-releasing hormone (GnRH) neuronal system in male gray hamsters, Tscherskia triton. Hamsters were kept from birth under a long-day (LD: 16 h light and 8 h dark) photoperiod and at 23°C until the experiment began. The animals were then kept for 30 days under different photoperiods and ambient temperatures. After autopsy, immunocytochemical and semiquantitative reverse transcription-polymerase chain reaction analyses were performed on the hamster forebrain. Exposure to a short-day (SD: 8 h light and 16 h dark) photoperiod at 23°C induced a functional decline in the GnRH neuronal system (increased neuron number and immunoreactivity, and reduced cell size). LD with low ambient temperature (7°C) did not influence the GnRH neuronal system. In contrast, SD with low ambient temperature enhanced the SD-induced functional decline in GnRH neurons. Changes in immunoreactivity of GnRH fibers in the median eminence, their release site, were also similar to those in neuronal perikarya. Significant differences in GnRH gene expression were detected in the SD only group and the SD with low ambient temperature group. However, no differences were detected between the two groups. These findings suggest that low ambient temperature enhanced the SD-induced decline in GnRH release from nerve terminals, but was not a crucial factor in the synthesis of this decapeptide.