The action of estrogen on target organs has been actively studied with the discovery of estrogen receptor (ER) beta. This study was carried out to examine the expression of ERalpha and ERbeta in the uterus and the vagina of immature Sprague-Dawley rats treated with 17-ethinyl estradiol (EE). Twenty days old rats were subcutaneously treated with EE at the doses of 0 (vehicle control), 0.03, 0.3, 1.0, 3.0, and 10.0 microg/kg/day for three consecutive days. The treatment of EE at the doses of 0.3, 1.0, 3.0 and 10.0 microg/kg/day significantly increased the weights of the uterus and vagina of rats (p<0.01) and retained fluid in the uterus of rats. At the high doses of 3.0 and 10.0 microg/kg/day, the treatment of EE caused an increase in the uterine height, hypertrophy, and a decrease in the expression of ERalpha and ERbeta in the uterine luminal and glandular epithelium. The treatment of EE at the doses of 3.0 and 10.0 microg/kg/day also caused cornification and a decrease in the expression of ERalpha and ERbeta in the vaginal epithelium. These results suggest that the EE treatment decrease the expression of ERalpha and ERbeta in the uterus and vagina of immature rats and that may be associated with the morphological changes such as increase in the uterine height, hypertrophy of the uterine epithelium, and cornification of the vagina.