Effect of vitrification on global gene expression dynamics of bovine elongating embryos.
Effect of vitrification on global gene expression dynamics of bovine elongating embryos.
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玻璃化冷冻对牛延长胚胎整体基因表达动态的影响。
DOI：
10.1071/rd20285
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发表时间：
2021-03
期刊：
影响因子：
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通讯作者：
Jiang Z
中科院分区：
文献类型：
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作者：
Gutierrez-Castillo E;Ming H;Foster B;Gatenby L;Mak CK;Pinto C;Bondioli K;Jiang Z
胚胎玻璃化冷冻涉及在冷冻保存过程中冷却和升温期间暴露于高浓度的冷冻保护剂和渗透应力。这些因素中有许多可能影响基因表达。在这项研究中，在体外生产的牛胚胎在囊胚阶段进行玻璃化冷冻。分别使用4只受体移植非玻璃化冷冻（n = 80）和玻璃化冷冻（n = 80）胚胎。通过子宫冲洗，共回收了12个非玻璃化冷冻和9个玻璃化冷冻的第14天（D14）存活胚胎。对来自玻璃化和新鲜回收的D14胚胎的整个胚胎或分离的滋养外胚层（TE）的RNA-seq分析显示，作为玻璃化的结果，在胚胎和TE分离物中分别共有927和4376个基因表达改变。此外，我们发现671和61基因通常上调或下调在玻璃化的整个胚胎和TE。玻璃化通常上调的途径包括上皮粘附连接，沉默调节蛋白信号传导，生殖细胞-支持细胞连接，ATM信号传导，NER和蛋白质泛素化途径。常见的下调途径包括EIF 2信号传导、氧化磷酸化、线粒体功能障碍、eIF 4和p70 S6 K信号传导的调节以及mTOR信号传导途径。我们的分析确定了影响胚胎发育能力的特定途径和涉及的特定基因表达模式，这些对冷冻保存很重要。
Embryo vitrification involves exposure to high concentrations of cryoprotectants and osmotic stress during cooling and warming in the cryopreservation process. Many of these factors can potentially affect gene expression. In this study, in vitro-produced bovine embryos at the blastocyst stage were subjected to vitrification. Four recipients each were used for transferring non-vitrified (n = 80) and vitrified (n = 80) embryos. A total of 12 non-vitrified and 9 vitrified viable day-14 (D14) embryos were recovered by uterine flushing. RNA-seq analysis of the whole embryo or isolated trophectoderm (TE) from vitrified and fresh recovered D14 embryos revealed a total of 927 and 4376 genes with changed expression in embryos and TE isolates, respectively, as a result of vitrification. In addition, we found 671 and 61 genes commonly up- or downregulated in both vitrified whole embryos and TE. Commonly upregulated pathways by vitrification included epithelial adherens junctions, sirtuin signalling, germ cell–sertoli cell junction, ATM signalling, NER and protein ubiquitination pathways. The commonly downregulated pathways included EIF2 signalling, oxidative phosphorylation, mitochondrial dysfunction, regulation of eIF4 and p70S6K signalling and mTOR signalling pathways. Our analysis identified specific pathways and implicated specific gene expression patterns affecting embryo developmental competence that are important to cryopreservation.