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The effect of ovariectomy, progesterone and prolactin treatment on the action of prostaglandin F₂ (PGF₂) was determined in pregnant rats. PGF₂ (150 µg x 2) injected i.p. on day 19 or 18 of pregnancy induced lactogenesis about 25 h later and abortion on days 20 and 21 of pregnancy. Treatment with PGF₂ (100 µg x 2 or 50 µg x 2) on day 19 induced lactogenesis around 22 or 38 h later, respectively, and abortion on day 21. PGF₂ treatment on day 17 was less effective. Unilateral ovariectomy on day 17 of pregnancy induced lactogenesis 32 h later but not abortion. PGF₂ (150 µg x 2) given on the day of surgery advanced lactogenesis 12 h and rats aborted on day 19. Bilateral ovariectomy on day 17 induced abortion between days 20 to 21, but if a single dose of PGF₂ (300 µg) was injected on day 18, all the ovariectomized rats aborted on day 19. Progesterone (10 mg) injected into rats treated with PGF₂ (150 µg x 2) on day 18, prevented abortion and delayed lactogenesis. Prolactin (1 mg x 4) treatment delayed only abortion. Serum prolactin levels were significantly higher 12 h after the last dose of PGF₂ (150 µg x 2) in rats treated on days 17, 18 or 19 of pregnancy. Pretreatment with progesterone prevented the rise in prolactin concentration. These results suggest that the lactogenic and abortive action of PGF₂ may be dependent on the uterine and plasma concentration of progesterone.