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Integrated Analysis of Quantitative Proteome and Transcriptional Profiles Reveals the Dynamic Function of Maternally Expressed Proteins After Parthenogenetic Activation of Buffalo Oocyte

机译:定量蛋白质组和转录谱的综合分析揭示了水牛卵母细胞孤雌激活后母体表达蛋白的动态功能。

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摘要

Maternal-effect genes are especially critical for early embryonic development after fertilization and until massive activation of the embryonic genome occurs. By applying a tandem mass tag (TMT)-labeled quantitative proteomics combined with RNA sequencing approach, the proteome of the buffalo was quantitatively analyzed during parthenogenesis of mature oocytes and the two-cell stage embryo. Of 1908 quantified proteins, 123 differed significantly. The transcriptome was analyzed eight stages (GV, MII, 2-cell, 4-cell, 8-cell, 16-cell, morula, blastocyst) of Buffalo using the RNA sequencing approach, and a total of 3567 unique genes were identified to be differently expressed between all consecutive stages of pre-implantation development. Validation of proteomics results (TUBB3, CTNNA1, CDH3, MAP2K1), which are involved in tight junction and gap junction, revealing that the maternal expression of the proteins possibly plays a role in the formation of cellular junctions firstly after parthenogenetic activation. Correlation and hierarchical analyses of transcriptional profiles and the expression of NPM2 and NLRP5 mRNA of buffalo in vitro developed oocytes and parthenogenetic embryos indicated that the “maternal-to-zygotic transition” (MZT) process might exist in the model of parthenogenesis, which is similar to a normally fertilized embryo, and may occur between the 8-cell to 16-cell stage. These data provide a rich resource for further studies on maternal proteins and genes and are conducive to improving nuclear transfer technology.
机译:母本效应基因对于受精后直至胚胎基因组大量激活之前的早期胚胎发育特别重要。通过应用串联质量标签(TMT)标记的定量蛋白质组学与RNA测序方法相结合,在成熟卵母细胞和两细胞阶段胚胎单性生殖过程中对水牛的蛋白质组进行了定量分析。在1908种定量蛋白质中,有123种差异显着。使用RNA测序方法对Buffalo的8个阶段(GV,MII,2细胞,4细胞,8细胞,16细胞,桑ula,胚泡)的转录组进行了分析,共鉴定出3567个独特基因在植入前发展的所有连续阶段之间表达不同。蛋白质组学结果的验证(TUBB3,CTNNA1,CDH3,MAP2K1)涉及紧密连接和间隙连接,揭示了蛋白质的母体表达可能首先在孤雌生殖激活后在细胞连接的形成中起作用。水牛体外发育卵母细胞和孤雌生殖胚胎的转录谱以及NPM2和NLRP5 mRNA的表达的相关性和层次分析表明,孤雌生殖模型中可能存在“母体-合子转变”(MZT)过程,这一点相似到正常受精的胚胎,可能发生在8细胞到16细胞阶段之间。这些数据为进一步研究母体蛋白质和基因提供了丰富的资源,有利于改进核移植技术。

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