Immunological analysis of the biochemical properties of the uterine estrogen receptor.
Immunological analysis of the biochemical properties of the uterine estrogen receptor.
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子宫雌激素受体生化特性的免疫学分析。
DOI：
10.1095/biolreprod40.6.1275
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发表时间：
1989
影响因子：
3.6
通讯作者：
K. Korach
中科院分区：
文献类型：
--
作者：
Shuji Yamashita;K. Korach
用雌激素受体（ER）单克隆抗体H-222进行免疫组化和免疫印迹分析，以阐明雌激素受体（ER）的“核转位”现象，阐明雌激素受体的主要核结合位点，以及评价雌激素受体与其核结合位点在去卵巢成年小鼠子宫中的结合力。ER在上皮细胞、间质细胞和平滑肌细胞中均定位于核内。从注射生理盐水、17 β-雌二醇（E2）、雌三醇（E3）和己烯雌酚（DES）的动物制备的子宫组织在切片前固定（预固定法）和使用冷冻切片时表现出几乎相同的ER免疫染色。另一方面，当新鲜冷冻切片与各种溶液孵育之前或之后固定（后固定法），然后用各种盐溶液处理时，在盐水注射和盐水处理的动物之间ER的免疫染色中观察到更大的差异。与PBS（含有0.16 M NaCl的0.01 M磷酸盐缓冲液，pH 7.2）孵育后，对照动物的ER免疫染色较低，而注射激素的小鼠的子宫组织在这种处理后显示出强烈的核免疫染色。在用0.4 M KCl或0.5 M NaCl处理后，注射过雌二醇的动物和对照动物的子宫中的免疫染色被完全消除。DNA酶治疗引起的免疫染色的ER几乎完全丧失，然而，RNA酶消化略有增加免疫反应性在E2注射和对照组动物。使用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳和Western印迹技术的定量分析表明，孵育后的组织切片与PBS，0.4 M KCl，或DNA酶，60%，10%，和30%的ER分别存在于未孵育的部分相比，ER的量。这些研究结果表明以下子宫组织中的ER;核占有率是由于细胞核中被占据和未被占据的受体之间的差异亲和力而发生的现象;激素治疗后，受体水平在细胞核中不会波动到结合试验所证明的程度;并且在免疫组织化学分析中检测到的ER的性质与生化研究中观察到的相同。
Immunohistochemical and immunochemical analysis using Western blot techniques were carried out with estrogen receptor (ER) monoclonal antibody H-222 to 1) clarify the "nuclear translocation" phenomenon of ER, 2) elucidate the primary nuclear binding site of ER, and 3) to evaluate the binding force between ER and its nuclear binding site in the uterus of ovariectomized adult mice. Exclusive nuclear localization of ER was recognized in the epithelial cells, stroma cells, and smooth muscle cells. Uterine tissues prepared from animals injected with saline, 17 beta-estradiol (E2), estriol (E3), and diethylstilbestrol (DES) exhibited almost the same ER immunostaining when they were fixed prior to sectioning (prefixation method) and frozen sections were used. On the other hand, when fresh-frozen sections were fixed before or after incubation with various solutions (postfixation method) and then treated with various salt solutions, greater differences were seen in immunostaining of ER between saline-injected and hormone-treated animals. Immunostaining of ER in control animals was low after incubation with PBS (0.01 M phosphate buffer containing 0.16 M NaCl, pH 7.2), whereas uterine tissue from hormone-injected mice showed strong nuclear immunostaining after this treatment. After treatment with 0.4 M KCl or 0.5 M NaCl, immunostaining in the uterus of both hormone-injected and control animals was completely abolished. DNase treatment caused an almost complete loss of immunostaining of ER; however, RNase digestion slightly increased immunoreactivity in both E2-injected and control animals. Quantitative analysis using sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blot techniques showed that after incubation of tissue sections for 30 min with PBS, 0.4 M KCl, or DNase, 60%, 10%, and 30% of ER were present, respectively, compared to amount of ER present in unincubated sections. These findings suggest the following for the ER in uterine tissue; nuclear occupancy is a phenomenon that occurs due to a differential affinity between occupied and unoccupied receptors in the nucleus; after hormone treatment, the receptor levels do not fluctuate in the nucleus to the extent demonstrated by binding assays; and the properties of the ER detected in the immunohistochemical analysis are identical to those observed in biochemical studies.
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核基质中的雌激素受体：使用单克隆抗受体抗体直接演示。
影响因子：
4.8
作者：
Alexander,RB;Greene,GL;Barrack,ER
通讯作者：
Barrack,ER
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
DOI：
10.1016/0022-4731(84)90188-2
发表时间：
1984-01-01
影响因子：
4.1
作者：
GREENE, GL;SOBEL, NB;JENSEN, EV
通讯作者：
JENSEN, EV
使用单克隆抗体对大鼠脑、垂体和子宫中雌激素受体进行免疫组织化学定位。
DOI：
10.1016/0022-4731(86)90111-1
发表时间：
1986
期刊：
Journal of steroid biochemistry
影响因子：
--
作者：
Sar,M;Parikh,I
通讯作者：
Parikh,I
影响因子：
4.8
作者：
JORDAN, VC;TATE, AC;WELSHONS, WV
通讯作者：
WELSHONS, WV