Immunodetection of Estrogen Receptors in Fetal and Neonatal Female Mouse Reproductive Tracts^*

Abstract

An immunocytochemical assay for estrogen receptor (ER) was used to study the distribution of receptor in fetal and immature female mouse reproductive tracts. Immunoblots confirmed that a single band, the size of the ER, immunostained in extracts from day 15 and 17 fetal reproductive tracts. Staining was observed over nuclei of epithelial cells of the Mullerian duct and over nuclei of cells of the developing connective tissue (mesenchymal cells) of the reproductive tract on fetal day 15. By day 17 when a primitive uterus could be distinguished, ERs were detected in nuclei of mesenchymal cells, but in only a small portion of epithelial cells. A different pattern of immunocytochemical staining was observed in uteri from animals killed on the day of birth; cells of the connective tissue contained ER, but the epithelial cells did not. By 4 or 6 days after birth, more nuclei in the connective tissue stained for ER with a greater intensity compared to nuclear staining in epithelial cells. ERs were detectable in nuclei of both uterine epithelial cells and connective tissue cells on days 10 and 19 after birth.

Estrogen receptor (ER) and its messenger ribonucleic acid expression in the genital tract of female mice exposed neonatally to tamoxifen and diethylstilbestrol

“…Although the uterus of early postnatal mice does not express ER protein in epithelial cells until at least the third postnatal day Bigsby et al, 1990;Greco et al, 1991;Sato et al, 19921, a single injection of E, or DES on day 0 induces ER protein expression in uterine epithelial cells in mice (Yamashita et al, 1990;Sato et al, 1992). In the present study, ER mRNA and ER protein in uterine epithelial cells were induced 4 h and 12 h later, respectively, by a single injection of DES on day 0.…”

“…For studies of immunoblots, uterus from adult ovariectomized mice were minced and extracted in 10 mM Tris and 2% sodium dodecyl sulfate (SDS) buffer by sonication and passed through a 25G needle as described previously (Greco et al, 1991). Proteins were analyzed by 7.5% SDS-polyacrylamide gel electrophoresis and transferred to PVDF membranes that were incubated in Tris-buffered saline (TBS) (pH 7.5) containing 5% skim milk (Difco, Detroit, MI) a t room temperature for 1 h, then treated with anti-ER monoclonal antibody (Abbott Laboratories, Chicago, IL) or nonimmune IgG (Abbott) at 1 5 0 dilution overnight.…”

“…et Kamashita et al, 1989;Bigsby et al, 1990;Greco et al, 1991;Sat0 et al, 19921, but a single injection of 17P-estradiol (E,) or diethylstilbestrol (DES) on the day of birth induces ER expression (Taguchi et al, 1988;Sat0 el;Yamashita et al, 1990). However, in the uterus of adult ovariectomized rats, ER mRNA levels were decreased by E, (Shupnik et al, 1989;Medlock et al, 1991).…”

Ontogeny of oestrogen receptor alpha in gonads and sex ducts of fetal and newborn mice

“…However, these studies have limitations in histological interpretation as a result of the use of either autoradiography (Stumpf et al, 1980;Schleicher et al, 1984;Holderegger and Keefer, 1986;Cooke et al, 1991) or frozen sections (Greco et al, 1991(Greco et al, , 1992. Furthermore, these studies do not discriminate between ER-α and ER-β.…”

Reduction of Primordial Follicles Caused by Maternal Treatment with Busulfan Promotes Endometrial Adenocarcinoma Development in Donryu Rats

“…In addition, estradiol receptor mediated responsiveness plays an important role for the uterine adenocarcinoma development in rats [1] and women [2], although estrogen receptors were expressed only in a few areas of the epithelial cells in the fetal uterus on day 15 of gestation, and the receptor mediated responsiveness was absent in the development of the prenatal female reproductive tract in mice and rats [33][34][35]. Busulfan exerts any estrogenic activity.…”