Ontogenetic expression of progesterone receptor (PR) and effect of estrogens on PR expression in the fetal female rat reproductive tract were investigated. To evaluate ontogenetic PR expression, female reproductive tract from untreated fetuses was examined on gestational days (GD) 15.5, 17.5, 19.5 and 21.5. To evaluate estrogen effects, pregnant rats were injected once per day with oil, 17beta-estradiol (E(2)) or diethylstilbestrol (DES) from GD 15 through 21. Female fetuses were prepared for real-time reverse-transcription polymerase chain reaction (RT-PCR) or immunohistochemistry for PR. Increase in PR mRNA expression was detected in the Müllerian duct on GD 21.5 compared to that on GDs 15.5 and 17.5 in untreated fetuses (P<0.05). Prenatal administration of E(2) or DES increased Müllerian PR mRNA levels by GD 21.5 compared with oil controls (P<0.01). To identify cell and region in which PR was expressed and up-regulated by E(2) and DES, localization was evaluated within three regions along the Müllerian duct axis which differentiate into oviduct, uterus and upper vagina in immunohistochemistry. In untreated fetuses, Müllerian epithelial PR immunoreactivity was weak on GDs 15.5 and 17.5, but then became moderate on GDs 19.5 and 21.5 in all three regions. These fetuses exhibited faint signals in Müllerian mesenchymal PR immunoreactivity during gestational monitoring. Critically, Müllerian mesenchymal PR staining became intense after E(2) exposure in all three regions by GD 21.5, but no change was observed in Müllerian epithelial PR. Similarly, DES dramatically induced Müllerian mesenchymal PR in all regions by GD 21.5, and also enhanced proximal epithelial PR. On the other hand, middle and caudal epithelial PRs were reduced by DES. These affected mesenchymal and epithelial cells by DES were ER alpha immunopositive in the Müllerian duct, except for middle Müllerian epithelium. These findings clearly demonstrate cell-specific PR localization and region-specific effect of DES on PR in the developing rat Müllerian duct, and provide fundamental information critical for investigating the tissue-specific mechanisms underlying the prenatal response to estrogen receptor agonists.