Positive feedback loop between prostaglandin E2 and EGF-like factors is essential for sustainable activation of MAPK3/1 in cumulus cells during in vitro maturation of porcine cumulus oocyte complexes
Positive feedback loop between prostaglandin E2 and EGF-like factors is essential for sustainable activation of MAPK3/1 in cumulus cells during in vitro maturation of porcine cumulus oocyte complexes
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前列腺素 E2 和 EGF 样因子之间的正反馈环对于猪卵丘卵母细胞复合体体外成熟期间卵丘细胞中 MAPK3/1 的可持续激活至关重要
DOI：
10.1095/biolreprod.110.090092
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发表时间：
2011
影响因子：
3.6
通讯作者：
et al
中科院分区：
文献类型：
--
作者：
Yamashita;et al
期间 在猪卵丘-卵母细胞复合体（COCs）的体外成熟中，促卵泡激素（FSH）增加前列腺素E2（PGE 2）的产生和EGF样因子的表达水平。配体通过其特异性受体EP 2、EP 4或EGF受体通过自分泌系统作用于卵丘细胞。当每种途径被抑制剂抑制时，卵丘完全扩展和卵母细胞成熟不会发生。在这项研究中，我们研究了这两个途径之间的关系，在卵丘细胞的猪COCs。当卵母细胞与FSH共同培养时，FshrmRNA表达在5 h内即开始下降，而卵丘细胞Ptger 2、Ptger 4和Ptgs 2的表达在FSH共同培养5或10 h后明显增加。PTGS 2抑制剂NS 398不仅在任何培养时间点显著抑制卵丘细胞PGE 2的分泌，而且在10和20 h显著抑制卵丘细胞Areg、Ereg和Tace/Adam 17的表达，而在1或5 h不显著。在早期培养期间，MAPK 3和MAPK 1（MAPK 3/1）的磷酸化不受NS 398的影响;然而，在10和20小时，磷酸化被抑制的药物。此外，通过添加PGE 2或EGF克服了NS 398对MAPK 3/1磷酸化和靶基因表达的下调。FSH诱导的卵丘扩张和减数分裂进程的MII阶段也被抑制NS 398，而这些影响也克服了添加PGE 2或EGF。这些结果表明，PGE 2通过诱导EGF样因子参与了卵丘细胞中MAPK 3/1的持续激活，而EGF样因子是卵丘细胞扩展和减数分裂成熟所必需的。
During in vitro maturation of porcine cumulus-oocyte complexes (COCs), follicle-stimulating hormone (FSH) increases both prostaglandin E2 (PGE2) production and the expression levels of EGF-like factors. The ligands act on cumulus cells by the autocrine system due to their specific receptors, EP2, EP4, or EGF receptor. When each pathway is suppressed by inhibitors, complete cumulus expansion and oocyte maturation do not occur. In this study, we examined the relationship between both of these pathways in cumulus cells of porcine COCs. When COCs were cultured with FSH,FshrmRNA expression was immediately decreased within 5 h, whereasPtger2,Ptger4,andPtgs2expression levels were significantly increased in cumulus cells in the culture containing FSH for 5 or 10 h. The PTGS2 inhibitor NS398 significantly suppressed not only PGE2 secretion at any culture time point but alsoAreg,Ereg,andTace/Adam17expression in cumulus cells at 10 and 20 h but not at 1 or 5 h. During the early culture period, phosphorylation of MAPK3 and MAPK1 (MAPK3/1) was not affected by NS398; however, at 10 and 20 h, phosphorylation was suppressed by the drug. Furthermore, down-regulations of MAPK3/1 phosphorylation and expression of the target genes by NS398 was overcome by the addition of either PGE2 or EGF. FSH-induced cumulus expansion and meiotic progression to the MII stage were also suppressed by NS398, whereas these effects were also overcome by addition of either PGE2 or EGF. These results indicated that PGE2 is involved in the sustainable activation of MAPK3/1 in cumulus cells via the induction of EGF-like factor, which is required for cumulus expansion and meiotic maturation of porcine COCs.