Responsiveness of bovine cumulus-oocyte-complexes (COC) to porcine and recombinant human FSH, and the effect of COC quality on gonadotropin receptor and Cx43 marker gene mRNAs during maturation in vitro.
Responsiveness of bovine cumulus-oocyte-complexes (COC) to porcine and recombinant human FSH, and the effect of COC quality on gonadotropin receptor and Cx43 marker gene mRNAs during maturation in vitro.
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DOI：
10.1186/1477-7827-1-14
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发表时间：
2003-02-11
期刊：
影响因子：
--
通讯作者：
Watson, Andrew J
中科院分区：
文献类型：
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作者：
Calder, Michele D;Caveney, Anita N;Watson, Andrew J
体外囊胚阶段的发育明显少于体内，这可能是由于卵母细胞能力的缺陷。尽管大部分牛卵母细胞经历自发核成熟，但对于适当的细胞质成熟的要求却知之甚少。通常，在成熟培养基中添加超生理浓度的 FSH 和 LH，以改善卵丘扩张、受精和胚胎发育。因此，研究了不同浓度的猪卵泡刺激素（pFSH）和重组人卵泡刺激素（rhFSH）对体外牛卵丘扩张的影响。测定卵丘-卵母细胞复合物中 FSHr、LHr 和 Cx43 mRNA 的表达，以确定它们是否是卵母细胞能力的有用标记。在无血清培养基中，仅 1000 ng/ml pFSH 诱导显着的卵丘扩张，但在 10% 血清存在下，100 ng/ml pFSH 的效果被放大。相反，在没有血清的情况下，1 ng/ml rhFSH 会发生卵丘扩张。 FSHr mRNA 在成熟 0-6 小时时最高，然后丰度下降。同样，Cx43 mRNA 表达在 0-6 小时内最高，但在成熟 24 小时后下降。然而，LHr mRNA 的相对丰度在成熟 6-24 小时内没有变化。在质量较差的 COC 中检测到 FSHr、LHr 和 Cx43 mRNA 水平降低。总之，在无血清培养基中低剂量的 rhFSH 时，牛卵丘发生了扩张。综上所述，FSHr、LHr和Cx43 mRNA丰度反映了COC质量以及体外成熟过程中FSHr和Cx43 mRNA表达的变化；这些基因可能是卵母细胞发育能力的有用标记。
Substantially less development to the blastocyst stage occurs in vitro than in vivo and this may be due to deficiencies in oocyte competence. Although a large proportion of bovine oocytes undergo spontaneous nuclear maturation, less is known about requirements for proper cytoplasmic maturation. Commonly, supraphysiological concentrations of FSH and LH are added to maturation media to improve cumulus expansion, fertilization and embryonic development. Therefore, various concentrations of porcine FSH (pFSH) and recombinant human FSH (rhFSH) were investigated for their effect on bovine cumulus expansion in vitro. Expression of FSHr, LHr and Cx43 mRNAs was determined in cumulus-oocyte complexes to determine whether they would be useful markers of oocyte competence. In serum-free media, only 1000 ng/ml pFSH induced marked cumulus expansion, but the effect of 100 ng/ml pFSH was amplified in the presence of 10% serum. In contrast, cumulus expansion occurred with 1 ng/ml rhFSH in the absence of serum. FSHr mRNA was highest at 0-6 h of maturation, then abundance decreased. Similarly, Cx43 mRNA expression was highest from 0-6 h but decreased by 24 h of maturation. However, the relative abundance of LHr mRNA did not change from 6-24 h of maturation. Decreased levels of FSHr, LHr and Cx43 mRNAs were detected in COCs of poorer quality. In conclusion, expansion of bovine cumulus occurred at low doses of rhFSH in serum-free media. In summary, FSHr, LHr and Cx43 mRNA abundance reflects COC quality and FSHr and Cx43 mRNA expression changes during in vitro maturation; these genes may be useful markers of oocyte developmental competence.