Transcriptomic signature to oxidative stress exposure at the time of embryonic genome activation in bovine blastocysts
Transcriptomic signature to oxidative stress exposure at the time of embryonic genome activation in bovine blastocysts
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DOI：
10.1002/mrd.22162
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发表时间：
2013-04-01
影响因子：
2.5
通讯作者：
Sirard, Marc-Andre
中科院分区：
文献类型：
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作者：
Cagnone, Gael L. M.;Sirard, Marc-Andre
为了了解体外培养如何影响胚胎质量，我们分析了暴露于氧化应激条件增加后牛囊胚的存活率和整体基因表达。从第3天（816细胞期）开始，将两种促氧化剂分别加入体外培养基中，一种通过促进活性氧（ROS）产生（0.01 mM 2，2-偶氮双（2-脒基丙烷）二盐酸盐[AAPH]）在细胞外发挥作用，另一种通过抑制谷胱甘肽合成（0.4 mM丁硫基磺酰亚胺[BSO]）在细胞内发挥作用。然后对所得第7天囊胚进行转录组学分析。在文献中，这两种促氧化剂条件显示在一定比例的第8天囊胚中诱导延迟变性。在我们的实验中，没有可见的形态学差异，但AAPH倾向于降低囊胚率，而BSO显著降低囊胚率，表明对存活群体的差异影响。在转录组水平上，无论是促氧化剂暴露存活的胚泡显示氧化应激和炎症反应（ARRB 2），虽然AAPH诱导更高的干扰细胞内稳态（SERPINE 1）。然而，BSO档案的功能基因组学鉴定了与甘氨酸代谢和能量代谢（TPI 1）相关的基因的差异表达。这些差异特征可能表明AAPH群体中存在退化前囊胚（IGFBP 7），而BSO暴露将选择最具活力的个体（TKDP 1）。总之，这些结果说明了如何氧化破坏前附着的发展是与系统上调的几个代谢标志物。此外，它表明，更好的抗氧化剂耗尽的生存能力可能允许囊胚在压实后以更安静的代谢存活。摩尔Reprod. Dev. 9999：100。(c)2013 Wiley Periodicals，Inc.摩尔Reprod. Dev. 80：297314，2013. (c)2013 Wiley Periodicals，Inc.
In order to understand how in vitro culture affects embryonic quality, we analyzed survival and global gene expression in bovine blastocysts after exposure to increased oxidative stress conditions. Two pro-oxidant agents, one that acts extracellularly by promoting reactive oxygen species (ROS) production (0.01mM 2,2-azobis (2-amidinopropane) dihydrochloride [AAPH]) or another that acts intracellularly by inhibiting glutathione synthesis (0.4mM buthionine sulfoximine [BSO]) were added separately to in vitro culture media from Day 3 (816-cell stage) onward. Transcriptomic analysis was then performed on resulting Day-7 blastocysts. In the literature, these two pro-oxidant conditions were shown to induce delayed degeneration in a proportion of Day-8 blastocysts. In our experiment, no morphological difference was visible, but AAPH tended to decrease the blastocyst rate while BSO significantly reduced it, indicating a differential impact on the surviving population. At the transcriptomic level, blastocysts that survived either pro-oxidant exposure showed oxidative stress and an inflammatory response (ARRB2), although AAPH induced higher disturbances in cellular homeostasis (SERPINE1). Functional genomics of the BSO profile, however, identified differential expression of genes related to glycine metabolism and energy metabolism (TPI1). These differential features might be indicative of pre-degenerative blastocysts (IGFBP7) in the AAPH population whereas BSO exposure would select the most viable individuals (TKDP1). Together, these results illustrate how oxidative disruption of pre-attachment development is associated with systematic up-regulation of several metabolic markers. Moreover, it indicates that a better capacity to survive anti-oxidant depletion may allow for the survival of blastocysts with a quieter metabolism after compaction. Mol. Reprod. Dev. 9999:XXXX. (c) 2013 Wiley Periodicals, Inc. Mol. Reprod. Dev. 80: 297314, 2013. (c) 2013 Wiley Periodicals, Inc.