Center of Embryo Engineering and Technology, College of Animal Science and Technology, Nanjing Agricultural University, Nanjing, China.
Theriogenology. 2013 Sep 1;80(4):328-36. doi: 10.1016/j.theriogenology.2013.04.019. Epub 2013 Jun 7.
Oocyte aging severely decreases the quality of oocytes, which hampers fertilization and subsequent embryo development. In the present study, age-dependent molecular changes in goat oocytes were investigated. First, the quality of goat oocytes with various in vitro culture times (24, 30, 36, 48, and 60 hours) was evaluated on the basis of developmental rates of parthenogenetically activated embryos and apoptosis of cumulus cells (CCs). Second, relative gene expression of six genes (mitochondrial genes: PGC-1α and NRF-1; epigenetic modification genes: SNRPN and HAT1; mitotic spindle checkpoint protein: SMAD2; and hyaluronan synthase gene: HAS3) were analyzed during oocyte aging. Third, we further studied the changes of seven genes (PGC-1α and NRF-1; apoptotic-related genes: BAX and BCL2; hyaluronan synthase gene: HAS2; metabolism-related gene: STAR; and superoxide dismutase gene: SOD1) in CCs during oocyte aging. In these studies, the blastocyst rate gradually decreased and the number of apoptotic cells significantly increased as the culture time increased (P < 0.05). Moreover, relative gene expressions of PGC-1α, NRF-1 and SMAD2 significantly decreased from 24 to 36 hours (P < 0.05), whereas the levels of HAT1 and HAS3 slowly increased as culture was prolonged. Furthermore, the levels of PGC-1α, BCL2, HAS2 and SOD1 quickly reduced, and BAX significantly increased from 24 to 36 hours in aged CCs (P < 0.05). In conclusion, goat oocytes started to age at 30 hours in vitro culture, and gene expression patterns of oocytes and CCs significantly changed as the oocytes aged. Gene expression pattern changes in CCs may provide a convenient and effective way to detect oocyte aging without compromising oocyte integrity.
卵母细胞老化严重降低卵母细胞质量,从而阻碍受精和随后的胚胎发育。本研究旨在探讨山羊卵母细胞随年龄变化的分子变化。首先,根据孤雌激活胚胎的发育率和卵丘细胞(CCs)的凋亡情况,评估了具有不同体外培养时间(24、30、36、48 和 60 小时)的山羊卵母细胞的质量。其次,分析了卵母细胞老化过程中六个基因(线粒体基因:PGC-1α 和 NRF-1;表观遗传修饰基因:SNRPN 和 HAT1;有丝分裂纺锤体检查点蛋白:SMAD2;透明质酸合酶基因:HAS3)的相对基因表达。第三,我们进一步研究了卵母细胞老化过程中 CCs 中七个基因(PGC-1α 和 NRF-1;凋亡相关基因:BAX 和 BCL2;透明质酸合酶基因:HAS2;代谢相关基因:STAR;超氧化物歧化酶基因:SOD1)的变化。在这些研究中,随着培养时间的增加,囊胚率逐渐降低,凋亡细胞数量显著增加(P<0.05)。此外,PGC-1α、NRF-1 和 SMAD2 的相对基因表达从 24 小时到 36 小时显著降低(P<0.05),而 HAT1 和 HAS3 的水平随着培养时间的延长而缓慢增加。此外,老化 CCs 中 PGC-1α、BCL2、HAS2 和 SOD1 的水平在 24 小时至 36 小时内迅速降低,BAX 水平显著增加(P<0.05)。综上所述,山羊卵母细胞在体外培养 30 小时时开始衰老,卵母细胞和 CCs 的基因表达模式随卵母细胞衰老而显著改变。CCs 中的基因表达模式变化可能为不损害卵母细胞完整性的情况下检测卵母细胞衰老提供一种方便有效的方法。