Intracellular localization of the glucocorticoid receptor: evidence for cytoplasmic and nuclear localization.
Intracellular localization of the glucocorticoid receptor: evidence for cytoplasmic and nuclear localization.
复制标题
糖皮质激素受体的细胞内定位：细胞质和核定位的证据。
作者：
A. Wikström;Oddmund Bakke;S. Okret;Mikael Brönnegård;J
使用单特异性，单克隆抗体对糖皮质激素受体（GR），免疫细胞化学研究进行调查的GR在细胞内定位的存在或不存在的配体。采用所有测试的固定方法（多聚甲醛、乙酸乙醇溶液、布因固定剂和苯异醌PBS溶液），可以获得特异性GR染色。选择用多聚甲醛固定以进一步研究透化的效果，使用几种浓度的Triton X-100或皂苷。使用大鼠Rueber肝癌（H-4-II-E）和人子宫癌（NHIK 3025）细胞系以及来自正常大鼠的培养肝细胞。在双重免疫荧光染色实验中，通过使用针对具有已知位置的细胞组分（即核心核小体蛋白和微管蛋白）的抗体与抗GR抗体组合，测试固定和透化后不同细胞隔室的可及性。用间接过氧化物酶抗过氧化物酶技术或用异硫氰酸荧光素标记的第二抗体获得的特异性GR染色显示存在于细胞质和细胞核中。如果将单克隆抗体与90%纯GR制剂预孵育，则所有细胞室的染色被废除（过氧化物酶抗过氧化物酶）或减少（异硫氰酸荧光素）。这些发现与最近报道的免疫细胞化学研究相反，后者报道了雌激素和孕激素受体的严格核存在。因此，关于类固醇受体定位的一般化不能作出。此外，在体外模型进行了描述，其中地塞米松给药后的受体染色在H-4-II-E细胞的本地化的效果可以进行研究。
Using a monospecific, monoclonal antibody against the glucocorticoid receptor (GR), an immunocytochemical study was performed to investigate the intracellular localization of GR both in the presence or absence of ligand. With all fixation methods tested (paraformaldehyde, acetic acid in ethanol, Bouin's fixative, and bensochinone in PBS), it was possible to obtain specific GR staining. Fixation with paraformaldehyde was chosen for further studies on the effect of permeabilization, using several concentrations of Triton X-100 or saponin. A rat Rueber hepatoma (H-4-II-E) and a human uterus carcinoma (NHIK 3025) cell line were used as well as cultured hepatocytes from normal rat. The accessibility of the different cell compartments after fixation and permeabilization was tested for by using antibodies against cellular constituents with known locations (i.e. core-nucleosome proteins and tubulin), in combination with the anti-GR antibody in double immunofluorescence staining experiments. The specific GR stain obtained with the indirect peroxidase antiperoxidase technique or with fluorescein isothiocyanate-labeled second antibodies was shown to be present both in the cytoplasm and in the nucleus. Staining of all cellular compartments was abolished (peroxidase antiperoxidase) or diminished (fluorescein isothiocyanate) if the monoclonal antibody was preincubated with a 90% pure GR preparation. These findings are in contrast to recently reported immunocytochemical studies, where a strict nuclear existence of the estrogen and progestin receptors has been reported. Consequently, generalizations with regard to steroid receptor localization cannot be made. Furthermore, an in vitro model is described, where the effect of dexamethasone administration upon the localization of receptor staining in H-4-II-E cells can be studied.
使用单克隆雌性亲和蛋白抗体对雌激素受体进行免疫电子显微镜定位。
DOI：
10.1177/33.9.4020102
发表时间：
1985
期刊：
The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society
影响因子：
--
作者：
Press,MF;Nousek-Goebl,NA;Greene,GL
通讯作者：
Greene,GL
影响因子：
4.8
作者：
W. Welshons;B. Krummel;J. Gorski
通讯作者：
W. Welshons;B. Krummel;J. Gorski
糖皮质激素受体的单克隆抗体。
影响因子：
4.8
作者：
Grandics,P;Gasser,DL;Litwack,G
通讯作者：
Litwack,G