Progesterone is essential for maintenance of Tace/Adam17 mRNA expression, but not EGF-like factor, in cumulus cells, which enhances the EGF receptor signaling pathway during in vitro maturation of porcine COCs.
Progesterone is essential for maintenance of Tace/Adam17 mRNA expression, but not EGF-like factor, in cumulus cells, which enhances the EGF receptor signaling pathway during in vitro maturation of porcine COCs.
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DOI：
10.1262/jrd.09-199h
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发表时间：
2010-06
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通讯作者：
Y. Yamashita;Ikko Kawashima;Yosuke Gunji;M. Hishinuma;M. Shimada
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作者：
Y. Yamashita;Ikko Kawashima;Yosuke Gunji;M. Hishinuma;M. Shimada
在猪卵丘 - 卵母细胞复合体（COCs）的体外成熟过程中，孕酮由卵丘细胞分泌并作用于卵丘细胞自身，这是卵丘扩展和卵母细胞成熟所必需的。表皮生长因子样因子（双调蛋白，AREG；上皮调节蛋白，EREG）及其蛋白酶，肿瘤坏死因子α转换酶/去整合素金属蛋白酶17（TACE/ADAM17）也在卵丘细胞中表达，因此，可溶性表皮生长因子结构域作用于卵丘细胞上表达的表皮生长因子受体。在本研究中，我们检测了猪COCs卵丘细胞中孕酮功能与表皮生长因子样因子刺激之间的关系。当COCs在促卵泡激素（FSH）和促黄体生成素（LH）作用下培养时，卵丘细胞中会表达Areg、Ereg和Tace/Adam17。用孕酮受体（PGR）拮抗剂RU486处理，在任何培养时间点都不影响Areg和Ereg的mRNA表达水平。然而，在30或40小时时间点，RU486显著抑制了Tace/Adam17的mRNA水平、蛋白质水平及其活性。在培养20小时时，RU486不抑制细胞外调节蛋白激酶1/2（ERK1/2）的磷酸化以及靶基因（透明质酸合酶2（Has2）、肿瘤坏死因子α诱导蛋白6（Tnfaip6）和前列腺素内过氧化物合酶2（Ptgs2））的表达；然而，在40小时时，RU486显著下调卵丘细胞中ERK1/2的磷酸化以及靶基因的表达水平。此外，添加表皮生长因子可克服RU486在40小时产生的负面影响。这些结果表明，在猪COCs体外成熟过程中，卵丘细胞中TACE/ADAM17的水平受孕酮 - 孕酮受体途径调控。因此，我们得出结论：在猪COCs体外成熟过程中，孕酮诱导的TACE/ADAM17促使卵丘细胞产生可溶性表皮生长因子结构域，从而增强卵丘细胞的功能变化并促进卵母细胞的减数分裂成熟。
During in vitro maturation of porcine cumulus-oocyte complexes (COCs), progesterone was secreted from cumulus cells and acted on the cumulus cells themselves, which required for cumulus expansion and oocyte maturation. EGF-like factor (amphiregulin, AREG; epiregulin, EREG) and its protease, TACE/ADAM17, are also expressed in cumulus cells, and thereby, soluble EGF domain was acted on the EGF receptor expressed on cumulus cells. In this study, we examined the relationship between progesterone function and EGF-like factor stimuli in cumulus cells of porcine COCs. When COCs were cultured with FSH and LH, Areg, Ereg and Tace/Adam17 were expressed in cumulus cells. Treatment with a progesterone receptor (PGR) antagonist, RU486, did not affect the Areg and Ereg mRNA expression levels at any culture time points. However, the Tace/Adam17 mRNA level, protein level and its activity were significantly suppressed by RU486 at the 30 or 40 h time point. At 20 h of culture, phosphorylation of ERK1/2 and the expressions of target genes (Has2, Tnfaip6 and Ptgs2) were not suppressed by RU486; however, at 40 h, ERK1/2 phosphorylation and the target gene expression levels were significantly downregulated by RU486 in cumulus cells. Furthermore, the negative effects of RU486 at 40 h were overcome by the addition of EGF. These results indicated that the level of TACE/ADAM17 in cumulus cells was regulated by the progesterone-PGR pathway during in vitro maturation of porcine COCs. Therefore, we concluded that the progesterone-induced TACE/ADAM17 leads to production of soluble EGF domain from cumulus cells, which enhances functional changes of cumulus cells and progresses meiotic maturation of oocytes during in vitro maturation of porcine COCs.