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卵母细胞成熟过程中牛卵丘-卵母细胞复合体中胰岛素样生长因子系统成分及促性腺激素受体的表达

Expression of components of the insulin-like growth factor system and gonadotropin receptors in bovine cumulus-oocyte complexes during oocyte maturation.

作者信息

Nuttinck F, Charpigny G, Mermillod P, Loosfelt H, Meduri G, Freret S, Grimard B, Heyman Y

机构信息

UMR 1198 Biologie du Développement et Reproduction, INRA-ENVA, 94704 Maisons-Alfort, France.

出版信息

Domest Anim Endocrinol. 2004 Aug;27(2):179-95. doi: 10.1016/j.domaniend.2004.03.003.

DOI:10.1016/j.domaniend.2004.03.003
PMID:15219936
Abstract

IGF system expression has been largely explored in the bovine follicular wall whereas it remains poorly studied in the COC. Using semi-quantitative RT-PCR and Western blot analysis, we have investigated spatial and temporal expression of IGF-1, IGFR-1, IGFBP-2, IGFBP-4, as well as gonadotropin receptors in bovine COC during oocyte maturation. In addition, we have compared changes in the IGF system and FSHR expression during in vitro maturation in TCM199 alone or in the presence of 10 ng/ml of EGF. The transcripts for IGFR-1 and IGFBP-2 were detected in cumulus and germinal cells whereas IGF-1, IGFBP-4 and FSHR mRNA were restricted to cumulus cells. Topography of the IGF system and gonadotropin receptor expression within COC were unaffected by the maturation step. In contrast, levels of IGFBP-2 and FSHR expression decreased (P < 0.05) in matured COC. Under defined culture conditions, IGFBP-2 and FSHR mRNA expression remained at a high level in TCM199 alone and were significantly reduced (P < 0.05) in the presence of 10 ng/ml EGF after a 24 h period of in vitro maturation. In conclusion, our results demonstrate a cell-specific pattern of IGF system member gene expression within bovine COC suggesting interaction between the somatic and germinal compartments. In addition, synchronized changes in the pattern of COC IGFBP-2 and FSHR expression during oocyte maturation suggest possible synergistic actions between IGF-1 and FSH.

摘要

胰岛素样生长因子(IGF)系统的表达在牛卵泡壁中已得到大量研究,而在卵丘-卵母细胞复合体(COC)中的研究仍较少。我们运用半定量逆转录聚合酶链反应(RT-PCR)和蛋白质免疫印迹分析,研究了牛COC在卵母细胞成熟过程中IGF-1、IGFR-1、IGFBP-2、IGFBP-4以及促性腺激素受体的时空表达。此外,我们比较了单独使用TCM199或添加10 ng/ml表皮生长因子(EGF)进行体外成熟培养时,IGF系统和促卵泡激素受体(FSHR)表达的变化。在卵丘细胞和生殖细胞中检测到了IGFR-1和IGFBP-2的转录本,而IGF-1、IGFBP-4和FSHR的mRNA仅局限于卵丘细胞。COC内IGF系统和促性腺激素受体表达的拓扑结构不受成熟步骤的影响。相反,成熟COC中IGFBP-2和FSHR的表达水平降低(P < 0.05)。在特定培养条件下,单独使用TCM199时,IGFBP-2和FSHR的mRNA表达保持在较高水平,而在体外成熟24小时后添加10 ng/ml EGF时,其表达显著降低(P < 0.05)。总之,我们的结果表明牛COC内存在IGF系统成员基因表达的细胞特异性模式,提示体细胞和生殖细胞区室之间存在相互作用。此外,卵母细胞成熟过程中COC的IGFBP-2和FSHR表达模式的同步变化表明IGF-1和FSH之间可能存在协同作用。

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