Ovariectomized ewes were injected with either 10 or 30 mg of progesterone, administered either intramuscularly (im) or subcutaneously (sc) at 24, 8, 4 h before, or at the same time, as im injections of 1, 2 or 4 mg of estradiol-17β. Determinations of serum luteinizing hormone (LH), estrogen, and progesterone by radioimmunoassay (RIA) were conducted to study the effects of the treatments on LH release, and the time course of estrogen and progesterone serum concentrations following injection. In the first experiment, progesterone failed to block the estrogen-induced LH release in four of six ewes, and in all three ewes of the second experiment. Intramuscular injection of progesterone resulted in higher circulating progesterone concentrations than sc injection and reached peak levels at 2–4 h postinjection. Estrogen reached peak values at 30 min–2 h postinjection. The results showed that progesterone injected at either 4, 8 and 24 h before, or simultaneously with, a pharmacological dosage of estrogen did not consistently block the LH release in ovariectomized ewes.