Integrated Analysis of Quantitative Proteome and Transcriptional Profiles Reveals the Dynamic Function of Maternally Expressed Proteins After Parthenogenetic Activation of Buffalo Oocyte
Integrated Analysis of Quantitative Proteome and Transcriptional Profiles Reveals the Dynamic Function of Maternally Expressed Proteins After Parthenogenetic Activation of Buffalo Oocyte
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DOI：
10.1074/mcp.ra118.000556
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发表时间：
2018-10-01
影响因子：
7
通讯作者：
Zhang, Ming
中科院分区：
文献类型：
--
作者：
Chen, Fumei;Fu, Qiang;Zhang, Ming
母体效应基因对受精后的早期胚胎发育尤其重要，直到胚胎基因组发生大规模激活。采用串联质量标签（TMT）标记的定量蛋白质组学技术结合RNA测序技术，对布法罗成熟卵母细胞和2细胞期胚胎孤雌发育过程中的蛋白质组进行了定量分析。在1908种定量蛋白质中，有123种差异显著。利用RNA测序方法分析了布法罗胚胎发育的8个阶段（GV、MII、2-细胞、4-细胞、8-细胞、16-细胞、桑椹胚、囊胚）的转录组，共鉴定出3567个独特基因在着床前发育的所有连续阶段之间差异表达。验证了蛋白质组学结果（TUBB 3、CTNNA 1、CDH 3、MAP 2K 1），这些蛋白质参与了紧密连接和间隙连接，揭示了这些蛋白质的母体表达可能在孤雌激活后首先形成细胞连接中起作用。通过对布法罗体外发育卵母细胞和孤雌发育胚胎的转录水平与NPM 2和NLRP 5 mRNA表达水平的相关性和层次分析，表明在孤雌发育模式中可能存在类似于正常受精胚胎的母合子转换（MZT）过程，可能发生在8-细胞至16-细胞阶段。这些数据为进一步研究母体蛋白和基因提供了丰富的资源，有利于提高核移植技术。
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.