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In order to study the regulation of primary spermatogonial (SPG) proliferation in the frog (Rana esculenta), mitotic and labelling indices of these cells were analysed, in vivo and in vitro, under different experimental conditions and periods of the year. Hypophysectomy, irrespective of the period of the year and independent of temperature, showed a remarkable negative influence on the mitotic or labelling index of the primary SPG. Replacement therapy with homologous pars distalis homogenate stimulated the proliferative activity, the stimulation being of significantly greater magnitude at 18°C than at 4°C. In parallel, mitotic index in vitro, in different periods of the year, after a 24-h incubation, was significantly higher at 20°C than at 8°C. At 2°C no 3H-thymidine labelling of the primary SPG was observed. Primary SPG labelling index in vitro increased with temperature, reaching the highest value at 15°C; it was, however, very low at 28°C. Under in vitro conditions FSH and LH stimulated primary SPG proliferation only when given together, but in vivo hypophysectomy stimulated SPG proliferation independently; GnRHa, thyroxine and prolactin were strongly stimulatory. The present in vitro data confirmed that testosterone acts synergistically with FSH-like substances to influence SPG proliferation. Unilateral castration rapidly increased the labelling index of the primary SPG in the remaining testis and this increased proliferative activity is assumed to be responsible for increased spermatogenetic activity and augmentation of testis mass later in time. It is suggested that temperature represents the constraint which controls the primary SPG responsiveness to hormonal factors.