Antibodies to the estrogen receptor-alpha modulate rapid prolactin release from rat pituitary tumor cells through plasma membrane estrogen receptors.
Antibodies to the estrogen receptor-alpha modulate rapid prolactin release from rat pituitary tumor cells through plasma membrane estrogen receptors.
复制标题
雌激素受体-α 的抗体通过质膜雌激素受体调节大鼠垂体肿瘤细胞催乳素的快速释放。
DOI：
10.1096/fasebj.14.1.157
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发表时间：
2000
期刊：
影响因子：
--
通讯作者：
Watson,CS
中科院分区：
文献类型：
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作者：
Norfleet,AM;Clarke,CH;Gametchu,B;Watson,CS
用抗雌激素受体α（ERα）抗体研究了ERα蛋白在雌激素刺激大鼠垂体GH 3/B6/F10细胞快速分泌催乳素（PRL）中的作用。将细胞暴露于1 nM 17β-雌二醇（E2）3或6 min后，PRL释放显著增加。当检测特异性结合ERα但太大而无法扩散到细胞中的ERα Ab在细胞膜上的活性时，靶向ERα铰链区序列的Ab R4以时间和浓度依赖性方式增加PRL释放。针对不同铰链区表位的Ab H151降低PRL释放并阻断E2的刺激作用。针对DNA结合结构域（H226）或羧基末端（C542）产生的Ab没有生物活性。当通过免疫细胞化学检查每种Ab对细胞表面ERα的识别时，除H151外，所有Ab在固定的细胞中均产生免疫染色。然而，在活细胞中，Ab H151而不是Ab R4阻断荧光标记的E2-BSA的膜结合。总之，数据表明质膜ERα蛋白介导雌激素刺激的GH 3/B6/F10细胞PRL释放。这些结果也可能传达有关ERα膜形式的构象敏感区域的信息，这些区域参与对雌激素的快速非基因组反应。Norfleet，A. M.，克拉克角，澳-地H、Gametchu，B.，沃森，C. S.抗雌激素受体-α抗体通过质膜雌激素受体调节大鼠垂体瘤细胞催乳素的快速释放
Antibodies (Abs) raised against the estrogen receptor-α (ERα) were used to investigate the role of ERα proteins located at the plasma membrane in mediating the rapid, estrogen-stimulated secretion of prolactin (PRL) from rat pituitary GH3/B6/F10 cells. Exposure of the cells to 1 nM 17β-estradiol (E2) significantly increased PRL release after 3 or 6 min. When ERα Abs that bind specifically to ERα but are too large to diffuse into cells were tested for activity at the cell membrane, Ab R4, targeted to an ERα hinge region sequence, increased PRL release in a time- and concentration-dependent fashion. Ab H151, directed against a different hinge region epitope, decreased PRL release and blocked the stimulatory action of E2. Abs raised against the DNA binding domain (H226) or the carboxyl terminus (C542) were not biologically active. When each Ab was examined for recognition of ERα on the cell surface by immunocytochemistry, all except H151 generated immunostaining in aldehyde-fixed cells. In live cells, however, Ab H151 but not Ab R4 blocked the membrane binding of fluorescently tagged E2-BSA. Overall, the data indicate that plasma membrane ERα proteins mediate estrogen-stimulated PRL release from GH3/B6/F10 cells. These results may also convey information about conformationally sensitive areas of the membrane form of ERα involved in rapid, nongenomic responses to estrogens.—Norfleet, A. M., Clarke, C. H., Gametchu, B., Watson, C. S. Antibodies to the estrogen receptor-α modulate rapid prolactin release from rat pituitary tumor cells through plasma membrane estrogen receptors.