Adverse effects of light at night are associated with human health problems and with changes in seasonal reproduction in several species. Owing to its role in the circadian timing system, melatonin production is suspected to mediate excess nocturnal light. To test this hypothesis, we examined the effect of light pollution on the timing of seasonal reproduction on a strict Malagasy long-day breeder, the nocturnal mouse lemur (Microcebus murinus). We randomly exposed 12 males in wintering sexual rest to moonlight or to a light-mimicking nocturnal streetlight for 5 weeks. We monitored urinary 6-sulfatoxymelatonin concentrations (aMT6s), plasma testosterone concentrations, and testis size, and we recorded daily rhythms of core temperature and locomotor activity. In males exposed to light pollution, we observed a significant decrease in urinary aMT6s concentrations associated with changes in daily rhythm profiles and with activation of reproductive function. These results showed that males entered spontaneous sexual recrudescence leading to a summer acclimatization state, which suggests that light at night disrupts perception of day length cues, leading to an inappropriate photoentrainment of seasonal rhythms.