Center of Natural and Human Sciences, Universidade Federal do ABC, Santo André, SP, Brazil.
Institute of Biomedical Sciences, Universidade de São Paulo, SP, Brazil.
Oxid Med Cell Longev. 2017;2017:1502489. doi: 10.1155/2017/1502489. Epub 2017 Oct 25.
High oxygen levels during culture (IVC) can induce oxidative stress through accumulation of reactive oxygen species (ROS), negatively affecting embryo development. This study evaluated the effect of different O tensions during IVC on bovine blastocyst development and transcriptional status, considering transcription factors that play an essential role during early embryo development. For this purpose, embryos were produced by conventional protocols and cultured in two different oxygen tensions, physiological (5%) and atmospheric (20%). Expanded blastocysts were subjected to transcript quantitation analysis by RT-qPCR with Biomark™ HD System (Fluidigm, US), using 67 TaqMan assays specific for . Differences were observed in genes related to oxidation-reduction processes, DNA-dependent transcription factors, and factors related to important functional pathways for embryo development. Blastocyst rate was higher in the 5% O group and the number of cells was assessed, with the 5% O group having a higher number of cells. ROS concentration was evaluated, with a higher ROS presence in the 20% O group. Taken together, these results allow us to conclude that IVC of embryos at atmospheric O tension affects the expression of important transcription factors involved in multiple cell biology pathways that can affect embryo development, quality, and viability.
培养过程中(IVC)的高氧水平可通过活性氧(ROS)的积累引起氧化应激,从而对胚胎发育产生负面影响。本研究评估了 IVC 过程中不同氧张力对牛囊胚发育和转录状态的影响,同时考虑了在早期胚胎发育中发挥重要作用的转录因子。为此,通过常规方案产生胚胎,并在两种不同的氧张力(生理(5%)和大气(20%))下进行培养。使用针对 67 个 TaqMan 探针的 Biomark™ HD 系统(Fluidigm,美国)对扩展囊胚进行 RT-qPCR 转录定量分析,这些探针特异性针对 。在与氧化还原过程、DNA 依赖性转录因子以及与胚胎发育重要功能途径相关的因子相关的基因中观察到差异。5% O 组的囊胚率更高,并且评估了细胞数量,发现 5% O 组的细胞数量更多。还评估了 ROS 浓度,发现 20% O 组的 ROS 存在更高。综上所述,这些结果使我们能够得出结论,即在大气氧张力下进行 IVC 会影响参与多个细胞生物学途径的重要转录因子的表达,这些途径可能会影响胚胎发育、质量和活力。