Aneuploidy Detection and mtDNA Quantification in Bovine Embryos with Different Cleavage Onset Using a Next-Generation Sequencing-Based Protocol
Aneuploidy Detection and mtDNA Quantification in Bovine Embryos with Different Cleavage Onset Using a Next-Generation Sequencing-Based Protocol
复制标题
DOI：
10.1159/000452923
复制
发表时间：
2016-01-01
影响因子：
1.7
通讯作者：
Rubes, Jiri
中科院分区：
文献类型：
--
作者：
Hornak, Miroslav;Kubicek, David;Rubes, Jiri
牛胚胎现在通常用于农业系统，作为在世界范围内传播上级遗传学的一种手段，最终目的是养活不断增长的人口。进一步的调查，常见的人类体外受精胚胎，因此有优先权，以改善牛体外受精，因为有筛查非整倍体（染色体数目异常）。虽然非整倍体的发生率和后果在人类植入前胚胎中有很好的记录，但对其他动物的胚胎却不太了解。为了解决这个问题，我们评估了31个2细胞牛胚胎的非整倍体水平，这些胚胎来自早期和晚期分裂的受精卵。当代方法（全基因组扩增和下一代测序）允许对来自4-7岁供体的卵母细胞中的所有染色体进行非整倍性评估。我们还量化了所有评估的卵裂球中的线粒体DNA（mtDNA）水平，从而验证了它们与非整倍体水平相关的假设。在这一组牛胚胎中，非整倍体的总体发生率为41.1%，并与卵裂时间显著相关（晚期卵裂胚胎为77.8%，早期卵裂胚胎为31.8%）。此外，基于mtDNA序列读段计数，我们观察到在后期切割胚胎中mtDNA的中值量显著较低。这些发现进一步加强了牛系统作为人类IVF研究模型的使用。(C)2016 S. Karger AG，巴塞尔
Bovine embryos are now routinely used in agricultural systems as a means of disseminating superior genetics worldwide, ultimately with the aim of feeding an ever-growing population. Further investigations, common for human IVF embryos, thus have priority to improve cattle IVF, as has screening for aneuploidy (abnormal chromosome number). Although the incidence and consequences of aneuploidy are well documented in human preimplantation embryos, they are less well known for the embryos of other animals. To address this, we assessed aneuploidy levels in thirty-one 2-cell bovine embryos derived from early-and late-cleaving zygotes. Contemporary approaches (Whole Genome Amplification and next-generation sequencing) allowed aneuploidy assessment for all chromosomes in oocytes from donors aged 4-7 years. We also quantified mitochondrial DNA (mtDNA) levels in all blastomeres assessed, thereby testing the hypothesis that they are related to levels of aneuploidy. The overall incidence of aneuploidy in this cohort of bovine embryos was 41.1% and correlated significantly with the timing of cleavage (77.8% in late-cleaving vs. 31.8% in early-cleaving embryos). Moreover, based on mtDNA sequence read counts, we observed that the median mtDNA quantity is significantly lower in late-cleaving embryos. These findings further reinforce the use of the bovine system as a model for human IVF studies. (C) 2016 S. Karger AG, Basel