Epitope-dependent localization of estrogen receptorα, but not -β, in en face arterial endothelium
Epitope-dependent localization of estrogen receptorα, but not -β, in en face arterial endothelium
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DOI：
10.1152/ajpheart.00781.2002
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发表时间：
2003-04-01
影响因子：
4.8
通讯作者：
Moore, EDW
中科院分区：
文献类型：
--
作者：
Dan, P;Cheung, JCY;Moore, EDW
17 β-雌二醇（E-2）在内皮细胞中的快速、非基因组效应被认为是由膜相关雌激素受体（ER）引起的，而这在血管组织中还没有发现。为了鉴定膜ER，我们使用多种位点定向ER α或ER β抗体标记大鼠脑和冠状动脉内皮细胞。Western印迹显示一种新的55 kDa ER α亚型。用这些抗体和细胞核和小窝蛋白-1的标记物标记的细胞的三维图像用宽视野显微镜获得，去卷积，并进行数值分析。我们在细胞核和细胞周边发现ER α，其中三分之一与小窝蛋白-1共定位。受体定位依赖于抗体的表位。人卵巢表面上皮细胞产生类似的结果，但在大鼠子宫肌层，分布是表位独立的和核。ER β主要分布在核内，不依赖于表位。少量ER α与ER β共定位于细胞核内。两种动脉制剂的结果相同，对E-2不敏感。我们推测，不同的ER α构象在膜上，在细胞核中，和不同的细胞类型之间允许E-2触发细胞和位置特异性信号级联。
Rapid, nongenomic effects of 17beta-estradiol (E-2) in endothelial cells are postulated to arise from membrane-associated estrogen receptors (ERs), which have not been visualized in vascular tissue. To identify membrane ERs, we used multiple site-directed ERalpha or ERbeta antibodies to label en face rat cerebral and coronary arterial endothelia. Western blots revealed a novel 55-kDa ERalpha isoform. Three-dimensional images of cells labeled with these antibodies and markers for the nucleus and caveolin-1 were acquired with a wide-field microscope, deconvolved, and numerically analyzed. We found ERalpha in the nucleus and cell periphery, where one-third colocalized with caveolin-1. The receptor location was dependent on the epitope of the antibody. Human ovarian surface epithelium produced similar results; but in rat myometrium, the distribution was epitope independent and nuclear. ERbeta distribution was predominately intranuclear and epitope independent. A small amount of ERalpha colocalized with ERbeta within the nucleus. The results were identical in both arterial preparations and insensitive to E-2. We postulate that the different ERalpha conformations at the membrane, in the nucleus, and between different cell types allow E-2 to trigger cell- and location-specific signaling cascades.