Role of Pgrmc1 in estrogen maintenance of meiotic arrest in zebrafish oocytes through Gper/Egfr
Role of Pgrmc1 in estrogen maintenance of meiotic arrest in zebrafish oocytes through Gper/Egfr
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DOI：
10.1530/joe-14-0576
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发表时间：
2015-04-01
影响因子：
4
通讯作者：
Thomas, Peter
中科院分区：
文献类型：
--
作者：
Aizen, Joseph;Thomas, Peter
受体运输到细胞表面的调节及其对靶细胞对生长因子和激素的反应的影响仍然知之甚少。最近使用癌细胞获得的初步证据表明，表皮生长因子受体（EGFR）的表面表达依赖于其与孕酮受体膜组分1（PGRMC 1）的结合。雌激素对斑马鱼卵母细胞成熟（OM）的抑制作用是通过G蛋白偶联雌激素膜受体1（Gper 1）介导的，并涉及Egfr的激活。因此，在这项研究中，Pgrmc 1在正常细胞中的细胞表面表达和Egfr功能的潜在作用进行了研究，在这个体外OM模型的Egfr行动使用PGMRC 1信号传导抑制剂，AG 205。一个单一的类似于60 kDa的蛋白质带，这对应于Pgrmc 1二聚体的大小，检测到完全生长的卵母细胞的质膜上的蛋白质印迹。与PGRMC 1抑制剂AG 205（20 μ M）的共同治疗阻断了100 nM雌二醇-17 β和GPER激动剂G-1对裸露斑马鱼卵母细胞自发成熟的抑制作用。此外，通过与PGRMC 1抑制剂共孵育，可防止EGFR抑制剂AG 1478和AG 825（50 μ M）逆转这些雌激素对OM的作用。用AG 205抑制Pgrmc 1信号也引起卵母细胞膜上Egfr依赖性信号和Egfr表达的减少。这些结果表明，维护Pgrmc 1信号是必需的Egfr在斑马鱼卵母细胞膜上的表达和保存功能的Egfr在维持减数分裂阻滞通过雌激素激活Gper。
The regulation of receptor trafficking to the cell surface and its effect on responses of target cells to growth factors and hormones remain poorly understood. Initial evidence has been recently obtained using cancer cells that surface expression of the epidermal growth factor receptor (EGFR) is dependent on its association with progesterone receptor membrane component 1 (PGRMC1). Estrogen inhibition of oocyte maturation (OM) in zebrafish is mediated through G-protein-coupled estrogen membrane receptor 1 (Gper1) and involves activation of Egfr. Therefore, in this study, the potential roles of Pgrmc1 in the cell surface expression and functions of Egfr in normal cells were investigated in this in vitro OM model of Egfr action using an inhibitor of PGMRC1 signaling, AG205. A single similar to 60 kDa protein band, which corresponds to the size of the Pgrmc1 dimer, was detected on plasma membranes of fully grown oocytes by western blotting. Co-treatment with the PGRMC1 inhibitor AG205 (20 mu M) blocked the inhibitory effects of 100 nM estradiol-17 beta and the GPER agonist, G-1, on spontaneous maturation of denuded zebrafish oocytes. Moreover, reversal of these estrogen effects on OM by the EGFR inhibitors AG1478 and AG825 (50 mu M) was prevented by co-incubation with the PGRMC1 inhibitor. Inhibition of Pgrmc1 signaling with AG205 also caused a decrease in Egfr-dependent signaling and Egfr expression on oocyte cell membranes. These results indicate that maintenance of Pgrmc1 signaling is required for Egfr expression on zebrafish oocyte cell membranes and for conserving the functions of Egfr in maintaining meiotic arrest through estrogen activation of Gper.