G protein-coupled receptor 30 is an estrogen receptor in the plasma membrane
G protein-coupled receptor 30 is an estrogen receptor in the plasma membrane
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DOI：
10.1016/j.bbrc.2006.05.191
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发表时间：
2006-08-04
影响因子：
3.1
通讯作者：
Mizukami, Yoichi
中科院分区：
文献类型：
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作者：
Funakoshi, Takeshi;Yanai, Akie;Mizukami, Yoichi
最近，GPR30被报道为一种新型雌激素受体；然而，其细胞内定位仍存在争议。为了研究 GPR30 在体内的细胞内定位，我们针对 GPR30 上的不同表位制备了四种多克隆抗体。使用抗GPR30抗体和抗FLAG抗体的免疫细胞化学观察表明，转染后24小时FLAG-GPR30定位于质膜。在表达 FLAG-GPR30 的 HeLa 细胞中，用雌激素（17β-雌二醇或 E2）治疗会导致细胞内 Ca2+ 浓度 ([Ca2+](i)) 在 10 秒内升高。此外，E2在刺激后1小时内诱导GPR30从质膜易位至细胞质。免疫组织化学分析显示GPR30存在于CA2锥体神经元细胞的细胞表面。透射电子显微镜图像显示 GPR30 定位于与锥体细胞质膜相关的特定区域。这些数据表明，GPR30（雌激素的跨膜受体）定位于大鼠大脑海马CA2锥体神经元细胞的质膜。 (c) 2006 Elsevier Inc. 保留所有权利。
Recently, GPR30 was reported to be a novel estrogen receptor; however, its intracellular localization has remained controversial. To investigate the intracellular localization of GPR30 in vivo, we produced four kinds of polyclonal antibodies for distinct epitopes on GPR30. Immunocytochemical observations using anti-GPR30 antibody and anti-FLAG antibody show that FLAG-GPR30 localizes to the plasma membrane 24 h after transfection. Treatment with estrogen (17 beta-estradiol or E2) causes an elevation in the intracellular Ca2+ concentration ([Ca2+](i)) within 10 s in HeLa cells expressing FLAG-GPR30. In addition, E2 induces the translocation of GPR30 from the plasma membrane to the cytoplasm by I h after stimulation. Immunohistochemical analysis shows that GPR30 exists on the cell surface of CA2 pyramidal neuronal cells. The images on transmission electron microscopy show that GPR30 is localized to a particular region associated with the plasma membranes of the pyramidal cells. These data indicate that GPR30, a transmembrane receptor for estrogen, is localized to the plasma membrane of CA2 pyramidal neuronal cells of the hippocampus in rat brain. (c) 2006 Elsevier Inc. All rights reserved.