The influence of pubertal age on the differential effect of a familiar versus an unfamiliar environment on social interaction (SI) in pairs of male rats was evaluated. The decrease in SI induced by the unfamiliar environment in adult rats is considered an anxiety-related response. Intact male rats and male rats castrated at 19 days were tested for SI at 28, 35, and 60 days of age. The results revealed that in the intact rats, decreased SI in an unfamiliar environment was evident at 35 and 60 days but not at 28 days. The behavioral composition of the environment-induced response at 35 days was different from that measured at 60 days. Prepubertal castration prevented the decrease in SI in the unfamiliar environment at both 35 and 60 days. This study demonstrated the emergence with the onset of puberty of a specific behavioral response to an anxiogenic condition. Furthermore, the development of this environment-related behavioral response was influenced by gonadal secretion(s), suggesting the importance of gonadal function to the emergence of this response.