Yamashita S, Newbold R R, McLachlan J A, Korach K S
Receptor Biology Section, National Institute of Environmental Health Sciences, Research Triangle Park, North Carolina 27709.
Endocrinology. 1990 Nov;127(5):2456-63. doi: 10.1210/endo-127-5-2456.
We have examined the relationship between localization of estrogen receptor (ER) and selected cell responses induced by estrogen in the neonatal CD-1 mouse uterus. The following simultaneous staining techniques were used to determine whether only uterine epithelial cells with ER are capable of showing proliferative and secretory activities after estrogen treatment: 1) ER localization and [3H]thymidine incorporation using immunohistochemistry and autoradiography; and 2) immunohistochemical double staining of ER and an estrogen-induced secretory protein lactoferrin (LF). Uterine tissues from two strains (CD-1 and BALB/c) mice on day 4 of age showed detectable ER by immunostaining in both epithelial and stromal cells. Day 4 CD-1 mice received a single injection of diethylstilbestrol (DES; 20 micrograms/kg BW). The percentage of ER-immunostained uterine epithelial cells and the intensity of the staining rapidly increased from 6-36 h, indicating that estrogen stimulates the expression and detectability of ER during the course of hormone treatment. Twenty-eight and 81% of epithelial cells showed positive ER immunostaining 6 and 24 h, respectively, after the DES treatment. The intensity of ER-immunostained stromal cells, however, was slightly decreased by the treatment. DES administration elicited epithelial cell proliferation. Labeling indices of epithelial cells reached a maximum (approximately 44%) 12 h after an injection of DES, and a second small peak was recognized 24 h after the treatment. Labeling indices of positively ER-immunostained epithelial cells and those of negatively stained epithelial cells showed almost the same pattern for 6-18 h after treatment. The respective maximum values were 45% and 43%. When day 4 mice were given three daily injections of DES in saline and killed 12 and 24 h after the last injection, significant amounts of LF were recognized in the apical cytoplasm of the epithelial cells. The epithelial cells that showed LF immunostaining always exhibited ER immunostaining in the nuclei. These results suggest that exogenous estrogen elicits increased expression of ER, DNA synthesis, and cell proliferation in neonatal uterine epithelial cells associated with low levels of ER, while there was a direct relationship with the presence of ER in epithelial cells and the induction of LF.
我们研究了雌激素受体（ER）的定位与雌激素诱导新生CD-1小鼠子宫中特定细胞反应之间的关系。采用以下同步染色技术来确定是否只有具有ER的子宫上皮细胞在雌激素处理后能够表现出增殖和分泌活性：1）使用免疫组织化学和放射自显影技术进行ER定位和[3H]胸腺嘧啶核苷掺入实验；2）对ER和一种雌激素诱导的分泌蛋白乳铁蛋白（LF）进行免疫组织化学双重染色。4日龄的两种品系（CD-1和BALB/c）小鼠的子宫组织经免疫染色显示，上皮细胞和基质细胞中均可检测到ER。4日龄的CD-1小鼠单次注射己烯雌酚（DES；20微克/千克体重）。ER免疫染色的子宫上皮细胞百分比和染色强度在6 - 36小时内迅速增加，表明雌激素在激素处理过程中刺激了ER的表达和可检测性。DES处理后6小时和24小时，分别有28%和81%的上皮细胞显示ER免疫染色呈阳性。然而，处理后ER免疫染色的基质细胞强度略有下降。给予DES可引起上皮细胞增殖。注射DES后12小时，上皮细胞的标记指数达到最大值（约44%），处理后24小时出现第二个小峰值。处理后6 - 18小时，ER免疫染色阳性的上皮细胞和阴性染色的上皮细胞的标记指数显示出几乎相同的模式。各自的最大值分别为45%和43%。当4日龄小鼠每天在盐水中注射三次DES，并在最后一次注射后12小时和24小时处死时，在上皮细胞的顶端细胞质中可识别出大量的LF。显示LF免疫染色的上皮细胞在细胞核中总是呈现ER免疫染色。这些结果表明，外源性雌激素可诱导新生子宫上皮细胞中ER表达增加、DNA合成及细胞增殖，而这些上皮细胞中的ER水平较低，同时上皮细胞中ER的存在与LF的诱导之间存在直接关系。
