Sequential exposure of porcine cumulus cells to FSH and/or LH is critical for appropriate expression of steroidogenic and ovulation-related genes that impact oocyte maturation in vivo and in vitro
Sequential exposure of porcine cumulus cells to FSH and/or LH is critical for appropriate expression of steroidogenic and ovulation-related genes that impact oocyte maturation in vivo and in vitro
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DOI：
10.1530/rep-08-0074
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发表时间：
2008-07-01
期刊：
影响因子：
3.8
通讯作者：
Shimada, Masayuki
中科院分区：
文献类型：
--
作者：
Kawashima, Ikkou;Okazaki, Tetsuji;Shimada, Masayuki
在本研究中，我们收集了马绒毛膜促性腺激素和人绒毛膜促性腺激素处理后特定时间间隔内有腔卵泡的卵泡液、颗粒细胞和卵丘细胞。心电图组在促进颗粒细胞和绒毛细胞增殖的同时，也增加了雌激素的产生。心电图还诱导卵丘细胞表达LHCGR和PGR，黄体生成素/人绒毛膜促性腺激素释放峰出现前卵泡液中孕酮积聚。此外，孕酮和孕酮受体(PGR)在FSH诱导卵丘细胞表达LjqCGR中起关键作用。卵丘细胞LHCGR mRNA的表达与黄体生成素诱导前列腺素的产生、表皮生长因子(EGF)样因子的释放以及具有凝血酶反应素样重复序列1的去整合素和金属蛋白酶的表达有关，从而促进卵丘细胞卵母细胞复合体(COCs)的扩张和卵母细胞的成熟。基于PGR和LHCGR在卵丘细胞中独特的表达和调控，我们设计了一种新的猪COCs培养系统，在该系统中依次添加激素来模拟体内观察到的变化。取小的有腔卵泡的COCs，用FSH和雌二醇预培养10h，再加入孕酮10h，20h后，将COCs移至含有促黄体生成素、EGF和孕酮的新鲜培养液中。在此改良的COC培养体系中成熟的卵母细胞表现出更强的囊胚期发育能力。根据这些结果，我们得出结论，为了在培养中实现最佳的COC扩增和卵母细胞成熟，需要确定每个物种卵丘细胞中独特的基因表达模式，并利用这些模式来增加激素刺激的有效性。
in this study, we collected follicular fluid, granulosa cells, and cumulus cells from antral follicles at specific time intervals following equine chorionic gonadotropin (eCG) and human chorionic gonadotropin (hCG) treatment of gilts. The treatment with eCG increased the production of estrogen coordinately with up-regulated proliferation of granulosa and CLIMUIUS cells. eCG also induced the expression of LHCGR and PGR in cumulus cells and progesterone accumulation was detected in folficular fluid prior to the LH/hCG surge. Moreover, progesterone and progesterone receptor (PGR) were critical for FSH-induced LjqCGR expression in cumulus cells in culture. The expression of LHCGR mRNA in cumulus cells was associated with the ability of LH to induce prostaglandin production, release of epidermal growth factor (EGF)-like factors, and a disintegrin and metalloprotease with thrombospondin-like repeats 1 expression, promoting cumulus cell oocyte complexes (COCs) expansion and oocyte maturation. Based on the unique expression and regulation of PGR and LHCGR in cumulus cells, we designed a novel porcine COCs culture system in which hormones were added sequentially to mimic changes observed in vivo. Specifically, COCs from small antral follicles were pre-cultured with FSH and estradiol for 10 h at which time progesterone was added for another 10 h. After 20 h, COCs were moved to fresh medium containing LH, EGF, and progesterone. The oocytes matured in this revised COC culture system exhibited greater developmental competence to blastocyst stage. From these results, we conclude that to achieve optimal COC expansion and oocyte maturation in culture thE' unique gene expression patterns in cumulus cells of each species need to be characterized and used to increase the effectiveness of hormone stimulation.