Histological and steroidogenic changes within dominant ovarian follicles (DFs) undergoing atresia following systemic administration of oestradiol benzoate (ODB) were characterized in beef heifers. At 5.6 6 0.1 days after the onset of oestrus, heifers received 1 mg ODB i.m./500 kg body weight (ODB; n 5 15) or served as controls (n 5 15). Timing of treatment initiation was designated as hour (h) 0 on day (d) 0, and coincided with the presence of the DF of the first follicular wave (DF1). Within treatments, the DF1 was collected following ovariectomy in four animals at h 12, h 36 or after ultrasonic detection of a new wave (NW) of ovarian follicular development. In heifers of the NW groups (n 5 7 per treatment), blood samples were collected at intervals of 20 min for 12 h beginning at h 2 12, 0, 24 and 48 to characterize circulating LH patterns. Administration of ODB suppressed (P < 0.01) mean concentrations of LH at h 24 and h 48 by preventing (P < 0.05) the increase in LH pulse amplitude observed in controls, but had no effect on FSH. Follicular fluid (FF) concentrations of androgens and oestradiol were reduced at h 36 in the ODB-treated group. The diameter of the DF1 and the number of granulosa cell layers were also reduced in ODB-treated as compared with control heifers. Treatment differences were not observed in the proportion of apoptotic granulosa cells as assessed using the TUNEL assay method, and timing of a new wave of follicular development (d 4.6 6 0.2) was similar (P > 0.1) among treatments. A prominent characteristic of oestradiol-induced atresia of the DF1 of the oestrous cycle in heifers was a loss in oestrogenic function associated with reduced LH support. However, the timing of new follicular development may be influenced by a factor(s) other than the status of the DF undergoing oestradiol-induced atresia.Reproduction (2005) 129 611-620