Abstract In the present study, the enhancing effect of synthetic estrogen on ovarian carbonyl reductase, a new prostaglandin (PG)-metabolizing enzyme, was investigated, and the antagonistic effect of antiestrogen on this enhancement was examined in immature rats. Ovarian carbonyl reductase activity towards 13,14-dihydro-15-keto-PGF 2α (15KD-PGF 2α), 4-benzoylpyridine (4BP) and menadione was determined photometrically and radiochemically, and quantitation of ovarian carbonyl reductase content was performed by Western blot analysis. Diethylstilbestrol (DES) and hexestrol (HX) enhanced the increasing effect of human chorionic gonadotropin (hCG) on ovarian carbonyl reductase activity and content when these synthetic estrogens (0.2 mg/kg) were administered for 3 days from 26 days of age, before hCG treatment. On the other hand, tamoxifen, which inhibits the binding of estradiol to the estrogen receptor, significantly prevented estradiol (E 2) from enhancing the effect of hCG on ovarian carbonyl reductase. Furthermore, the ovarian carbonyl reductase activities towards the three substrates correlated well with the ovarian carbonyl reductase content. These results indicate that ovarian carbonyl reductase in immature rats may be regulated by a specific increase in the ovarian response to luteinizing hormone mediated by estrogen receptor.