The relationship between comceptus number and serum estradiol levels was studied, and the role of estrogen in the regulation of corpus luteum (CL) function was investigated in midpregnant rats. The number of conceptuses was adjusted to one, or 10 or more on day 7 of pregnancy. Serum estradiol levels, serum progesterone levels, and CL weights in rats bearing one conceptus were significantly lower than those in rats bearing 10 or more conceptuses on day 15 of pregnancy. However, administration of estradiol 0.1μg/day decreased serum progesterone levels and CL weights both in rats bearing one conceptus and in rats bearing 10 or more conceptuses. These results suggest that estrogen itself might be luteolytic in midpregnant rats. Some possible explanations of this luteolytic effect of estrogen are discussed.