Gee J M, Nicholson R I, Jasani B, Newman G R, Amselgruber W M
Breast Cancer Unit, Tenovus Institute for Cancer Research, University of Wales College of Medicine, Cardiff, United Kingdom.
J Histochem Cytochem. 1990 Jan;38(1):69-78. doi: 10.1177/38.1.1688451.
We have developed an immunocytochemical method to demonstrate estrogen receptor in hormone-sensitive tissues of the rat using a dinitrophenyl (DNP) hapten-labeled rat antihuman estrogen receptor monoclonal antibody (MAb), H222. Mouse IgM anti-DNP was used secondarily, followed by a DNP/peroxidase conjugate, diaminobenzidine/hydrogen peroxide chromogen, and silver intensification. This method was applied to tissues from intact female rats and showed that estrogen receptor was localized in the nuclei of the stromal and glandular components of the uterine endometrium. Reduced receptor staining was observed in the luminal epithelium, with minimal myometrial staining. Anterior pituitary glands showed heterogeneous immunostaining and ovaries expressed the receptor predominantly in the interstitial cells; fallopian tubes demonstrated substantial epithelial staining. Uteri from chemically castrated rats showed reduced estrogen receptor immunostaining in both stromal and luminal cells, whereas staining was enhanced in the glandular elements. Classical estrogen-unresponsive tissues (heart, lung, and spleen) were unstained. Antibody controls involved pre-blocking antibody recognition sites on the receptor with unlabeled antibodies to estrogen receptor (H222, H226, and D547), as well as use of an inappropriate DNP-labeled antibody to metallothionein. These controls illustrated the specific nature of the DNP-H222 binding.
我们开发了一种免疫细胞化学方法，使用二硝基苯基（DNP）半抗原标记的大鼠抗人雌激素受体单克隆抗体（MAb）H222来检测大鼠激素敏感组织中的雌激素受体。其次使用小鼠IgM抗DNP，然后是DNP/过氧化物酶缀合物、二氨基联苯胺/过氧化氢显色剂和银增强剂。该方法应用于完整雌性大鼠的组织，结果表明雌激素受体定位于子宫内膜基质和腺体成分的细胞核中。在腔上皮中观察到受体染色减少，子宫肌层染色最少。垂体前叶显示出异质性免疫染色，卵巢主要在间质细胞中表达受体；输卵管显示出大量上皮染色。化学去势大鼠的子宫在基质和腔细胞中雌激素受体免疫染色均减少，而腺体成分中的染色增强。典型的雌激素无反应组织（心脏、肺和脾脏）未染色。抗体对照包括用未标记的雌激素受体抗体（H222、H226和D547）预先封闭受体上的抗体识别位点，以及使用不适当的DNP标记的金属硫蛋白抗体。这些对照说明了DNP-H222结合的特异性。
