Aromatization and antioxidant strategies in the male gonads are important processes, which are involved in control of normal fertility. The objective of this study was to show whether luteinizing hormone (LH), prolactin (PRL), and insulin-like growth factor-I (IGF-I) as well as the length of photoperiod are able to exert an effect on aromatase expression, steroid hormone levels, and antioxidant concentrations in testes of bank voles, seasonally breeding rodents. Mature bank voles that were kept under short light cycles or long light cycles served as the animal model. Testicular sections were used for immunohistochemical visualization of aromatase expression, whereas testicular homogenates were used for radioimmunological measurement of steroids, biochemical determination of superoxide dismutase activity, total antioxidant status (TAS) and protein content. In the testes of hormone-treated voles a stronger immunostaining for aromatase was concurrent with the increase in testosterone and estradiol levels, and total antioxidant status compared with the controls. In contrast, there was a decrease in superoxide dismutase (SOD) activity. The strongest effect on aromatase immunoexpression and steroid hormone levels was detected after combined doses of LH and IGF-I, indicating a stimulatory role of these hormones on estrogen synthesis in the bank vole. An increase in total antioxidant status in testes of hormone-treated bank voles suggests the presence of testicular defense, whereas a decrease in superoxide dismutase activity indicates a protective role of administered hormones against toxic oxygen radicals. The present study also demonstrates a significant, photoperiod-dependent relationship between aromatization and antioxidant capacity in the testis of the bank vole.