Department of Animal Science, Tokyo University of Agriculture, Atsugi, Japan.
Mol Reprod Dev. 2013 Jul;80(7):508-21. doi: 10.1002/mrd.22187. Epub 2013 May 31.
In general, maternal age affects the quality of oocytes and embryos. The present study aimed to examine the features and age-associated gene expression profiles of bovine oocytes and embryos as well as to discover possible countermeasures against age-associated events. Comprehensive gene expression assays of germinal vesicle and metaphase II (MII)-stage oocytes and 8- to 16-cell-stage embryos were conducted using next-generation sequencing technology. The gene expression profiles of aged cows showed high expression of genes related to oxidative phosphorylation, eIF4 and p70S6K signaling, and mitochondrial dysfunction in MII-stage oocytes. Oocytes derived from aged cows, compared with those derived from their younger counterparts, exhibited high levels of abnormal fertilization and blastocysts with low total cell numbers. Levels of reactive oxygen species (ROS) and SIRT1 were higher in in vitro-matured oocytes derived from aged cows than in those derived from their younger counterparts. Supplementation of maturation medium with N-acetyl-cysteine (NAC), but not resveratrol, reduced the levels of ROS in the oocytes derived from cows of both age groups; however, resveratrol, but not NAC, improved the fertilization ratio. Conversely, EX 527, an inhibitor of SIRT1, increased the ratio of abnormal fertilization. In conclusion, gene expression profiles of oocytes and embryos derived from aged cows differ from those of oocytes and embryos derived from young cows; in particular, oocytes derived from aged cows show protein and mitochondrial dysfunction. In addition, activation of SIRT1 in oocytes may be a potential countermeasure against age-associated events in oocytes derived from aged cows.
一般来说,母体年龄会影响卵母细胞和胚胎的质量。本研究旨在研究牛卵母细胞和胚胎的特征以及与年龄相关的基因表达谱,并发现可能对抗与年龄相关的事件的对策。使用下一代测序技术对生发泡期和中期 II(MII)期卵母细胞和 8-16 细胞期胚胎进行了全面的基因表达分析。高龄奶牛的基因表达谱显示,MII 期卵母细胞中与氧化磷酸化、eIF4 和 p70S6K 信号以及线粒体功能障碍相关的基因表达较高。与年轻奶牛相比,来自高龄奶牛的卵母细胞受精异常率较高,囊胚的总细胞数较低。与年轻奶牛相比,来自高龄奶牛的体外成熟卵母细胞中的活性氧(ROS)和 SIRT1 水平较高。在成熟培养基中添加 N-乙酰半胱氨酸(NAC)而非白藜芦醇可降低来自两个年龄组奶牛的卵母细胞中的 ROS 水平;然而,只有白藜芦醇而非 NAC 可以提高受精率。相反,SIRT1 的抑制剂 EX 527 增加了异常受精的比例。总之,来自高龄奶牛的卵母细胞和胚胎的基因表达谱与来自年轻奶牛的卵母细胞和胚胎的基因表达谱不同;特别是,来自高龄奶牛的卵母细胞表现出蛋白质和线粒体功能障碍。此外,卵母细胞中 SIRT1 的激活可能是对抗来自高龄奶牛的卵母细胞中与年龄相关的事件的潜在对策。