The main objective of the study was to determine the rate at which Graafian follicles of sheep that had been treated with exogenous gonadotrophin acquire the ability to secrete oestrogen in vitro. Follicles were explanted from sheep 5 min to 24 h after injection of pregnant mare serum gonadotrophin (PMSG) and kept individually in culture for 7 days. The mean daily output of oestrogen by follicles from PMSG-treated sheep was higher than that secreted by follicles from untreated sheep. However, only a certain proportion of the follicles from each sheep secreted significant amounts of oestrogen in vitro; these follicles were called 'stimulated'. The proportion of stimulated follicles was 5% for control sheep, 20–30% for follicles explanted from sheep 5 min to 12 h after injection with PMSG, and 80% for follicles explanted from sheep that had been injected with PMSG 24 h previously. In the second part of the study, the pattern of oestrogen and progesterone secretion by stimulated follicles of different sizes explanted from PMSG-treated sheep at various stages of the oestrous cycle was determined. Up to the 14th day, oestrogen production in vitro by follicles over 4·5 mm in diameter reached a maximum 2 days after PMSG injection and decreased thereafter; progesterone production rose steadily as the oestrogen levels declined. In contrast, follicles of less than 4·5 mm diameter secreted considerable amounts of oestrogen for the first 5 days in culture, but produced only small quantities of progesterone. In follicles explanted on day 15, oestrogen secretion decreased steadily from the beginning of the culture period and was very low by the 4th day. Most follicles explanted at oestrus secreted only small amounts of oestrogen in vitro but secreted large amounts of progesterone.