buffalo parthenogenesis TMT LC-MS/MS

Oocytes act as the genetic vector for zygote reprogramming, and stores a large amount of material that is required to regulate embryogenesis. However, to date, there is no report on the dynamic changes of maternal proteins and genes that occur during the early stages of the embryo, particularly studies that use proteomic techniques in buffalos. Here, an integrated single-cell RNA sequencing transcriptomic and quantitative proteomic analysis were employed to systematically explore the dynamic function of maternally-expressed proteins in parthenogenesis model of buffalo.The proteome of the buffalo was quantitatively analyzed during parthenogenesis of mature oocytes and the two-cell stage embryo. Of 1,908 quantified proteins, 123 differed significantly. The transcriptome was analyzed 8 stages (GV, MII, 2-cell,4-cell,8-cell,16-cell,morula,blastocyst)of buffalo using the single-cell RNA sequencing, and a total of 3567 unique genes were identified to be differently expressed between all consecutive stages of pre-implantation development. Bioinformatics studies and validated results indicated that maternal expression of the proteins possibly plays a role in the formation of cellular junctions firstly after parthenogenetic activation and the “maternal-to-zygotic transition” (MZT) process exists during parthenogenesis and occur between the 8-cell to 16-cell stage.

卵母细胞作为受精卵重编程的遗传载体，储存了调控胚胎发生所需的大量物质。然而迄今为止，尚无关于胚胎早期阶段母源蛋白与基因动态变化的相关报道，尤其是在水牛中采用蛋白质组学技术（proteomic techniques）开展的相关研究。本研究整合单细胞RNA测序（single-cell RNA sequencing）转录组学与定量蛋白质组分析技术，系统探究水牛孤雌生殖（parthenogenesis）模型中母源表达蛋白的动态功能。我们对成熟卵母细胞孤雌生殖及二细胞期胚胎过程中的水牛蛋白质组进行了定量分析，在1908种定量检测到的蛋白质中，有123种存在显著差异。本研究通过单细胞RNA测序，对水牛胚胎发育的8个阶段（GV期、MII期、2细胞期、4细胞期、8细胞期、16细胞期、桑椹胚期、囊胚期）进行转录组分析，共鉴定出3567个在着床前发育所有连续阶段间差异表达的独特基因。生物信息学分析与验证结果表明，母源表达蛋白可能在孤雌激活后早期参与细胞连接的形成，且孤雌生殖过程中存在母源-合子转换（Maternal-to-Zygotic Transition, MZT）过程，该过程发生于8细胞期至16细胞期之间。