比較人工合成乙醯雌酚 (DES, diethylstilbesterol) 誘導幼鼠所產生之子宮液與正常成鼠於動情週期所產生之子宮液兩者蛋白質的成分,發現一個分子量為 14kDa 、等電點為 4.6 的蛋白於成鼠表現,但於 DES 誘導的幼鼠沒有表現。由 MALDI-TOF-TOF 質譜分析,證實此一蛋白質為 proline-rich acidic protein (PRAP) 。 於成鼠的動情週期中, PRAP mRNA 表現於動情期 (estrus) 和早期動情後期 (metestrus I) ,但於動情前期 (proestrus) 表現較少,於後期動情後期 (metestrus II) 和動情間期 (diestrus) 則快速遞減。 在未成熟的老鼠子宮並不會表現 PRAP mRNA 。利用免疫組織化學法檢視 PRAP 在子宮主要分布於子宮內膜的上皮細胞和腺體表皮細胞。 對未成熟的小鼠施予動情素 (17β-estradiol) 或動情素加上黃體素 (progesterone) 三天後,於子宮中檢視到發現少量 PRAP mRNA ,也於子宮液檢測到少量 PRAP 。但於六天後,施打動情素加上黃體素的動物會產生大量 PRAP mRNA 。 DES 和動情素雖然都能加速幼鼠子宮達到成熟並分泌子宮液,但表現 PRAP mRNA 的時間點卻不相同。 將新生之幼鼠或三週大幼鼠施以 DES ,待其成熟於動情期 (estrus) , 前類動物子宮之 PRAP mRNA 的表現與正常成鼠無明顯差異,後類動物子宮之 PRAP mRNA 和子宮液之 PRAP 蛋白於正常成鼠則有顯著的減少。
I compared the protein components in the uterine luminal fluid (ULF) collected from either the diethylstilbestrol (DES)-stimulate baby mouse or normal adult females at estrus stage. By the mass spectrometry, I identify 14- kDa protein to be proline-rich acidic protein (PRAP) that was undetectable in ULF of the DES-stimulated baby mice but appeared in ULF of adult in estrus stage. In the natural estrous cycle, the mRNA level of PRAP appeared in proestrus, increased markedly in estrus and early metestrus and declined sharply in the late metestrus and diestrus to an almost undetectable level. PRAP mRNA was not detected in the uterus of immature mice. Immunohistochemical staining patterns of the estrous uterine section showed that PRAP was immunolocaliezed in glandular and epithelial cells of endometrium. Although treatment of baby mice with DES for consecutive three days did not stimulated the PRAP expression in the uterine an injection of estradiol (E2) alone or E2 together with progesterone (P4) for consecutive three days stimulated of the PRAP expression. Six days after the first injection of E2 together with P4, the PRAP mRNA increased remarkedly. Neonatal DES exposure did not coursed a reduction but prepubertal DES exposure decreases remarkedly in the uterine expression of PRAP when the animals were sexually matured.