Institute of Farm Animal Genetics, 31535 Mariensee, Neustadt, Germany.
Reproduction. 2012 Sep;144(3):319-30. doi: 10.1530/REP-12-0134. Epub 2012 Jun 25.
The developmental capacity of oocytes from prepubertal cattle is reduced compared with their adult counterparts, and epigenetic mechanisms are thought to be involved herein. Here, we analyzed DNA methylation in three developmentally important, nonimprinted genes (SLC2A1, PRDX1, ZAR1) and two satellite sequences, i.e. 'bovine testis satellite I' (BTS) and 'Bos taurus alpha satellite I' (BTαS). In parallel, mRNA expression of the genes was determined by quantitative real-time PCR. Oocytes were retrieved from prepubertal calves and adult cows twice per week over a 3-week period by ultrasound-guided follicular aspiration after treatment with FSH and/or IGF1. Both immature and in vitro matured prepubertal and adult oocytes showed a distinct hypomethylation profile of the three genes without differences between the two types of donors. The methylation status of the BTS sequence changed according to the age and treatment while the methylation status of BTαS sequence remained largely unchanged across the different age and treatment groups. Relative transcript abundance of the selected genes was significantly different in immature and in vitro matured oocytes; only minor changes related to origin and treatment were observed. In conclusion, methylation levels of the investigated satellite sequences were high (>50%) in all groups and showed significant variation depending on the age, treatment, or in vitro maturation. To what extent this is involved in the acquisition of developmental competence of bovine oocytes needs further study.
与成年牛卵母细胞相比,未成年牛卵母细胞的发育能力降低,而表观遗传机制被认为与此有关。在这里,我们分析了三个发育重要的非印记基因(SLC2A1、PRDX1、ZAR1)和两个卫星序列,即“牛睾丸卫星 I”(BTS)和“Bos taurus alpha 卫星 I”(BTαS)的 DNA 甲基化。同时,通过定量实时 PCR 确定了基因的 mRNA 表达。通过超声引导的卵泡抽吸,在 FSH 和/或 IGF1 处理后,每周两次从未成年小牛和成年奶牛中回收卵母细胞,持续 3 周。未成熟和体外成熟的未成年和成年卵母细胞均表现出三个基因的明显低甲基化谱,供体类型之间无差异。BTS 序列的甲基化状态随年龄和处理而变化,而 BTαS 序列的甲基化状态在不同的年龄和处理组之间基本保持不变。所选基因的相对转录丰度在未成熟和体外成熟的卵母细胞中存在显著差异;仅观察到与来源和处理相关的微小变化。总之,在所研究的卫星序列中,甲基化水平在所有组中均较高(>50%),并且根据年龄、处理或体外成熟而显著变化。这在多大程度上参与了牛卵母细胞获得发育能力仍需要进一步研究。