Table_6_Global Transcriptomic Analyses Reveal Genes Involved in Conceptus Development During the Implantation Stages in Pigs.XLSX

Prenatal mortality remains a significant concern to the pig farming industry around the world. Spontaneous fetal loss ranging from 20 to 45% by term occur after fertilization, with most of the loss happening during the implantation period. Since the factors regulating the high mortality rates of early conceptus during implantation phases are poorly understood, we sought to analyze the overall gene expression changes during this period, and identify the molecular mechanisms involved in conceptus development. This work employed Illumina’s next-generation sequencing (RNA-Seq) and quantitative real-time PCR to analyze differentially expressed genes (DEGs). Soft clustering was subsequently used for the cluster analysis of gene expression. We identified 8236 DEGs in porcine conceptus at day 9, 12, and 15 of pregnancy. Annotation analysis of these genes revealed rRNA processing (GO:0006364), cell adhesion (GO:1904874), and heart development (GO:0007507), as the most significantly enriched biological processes at day 9, 12, and 15 of pregnancy, respectively. In addition, we found various genes, such as T-complex 1, RuvB-like AAA ATPase 2, connective tissue growth factor, integrins, interferon gamma, SLA-1, chemokine ligand 9, PAG-2, transforming growth factor beta receptor 1, and Annexin A2, that play essential roles in conceptus morphological development and implantation in pigs. Furthermore, we investigated the function of PAG-2 in vitro and found that PAG-2 can inhibit trophoblast cell proliferation and migration. Our analysis provides a valuable resource for understanding the mechanisms of conceptus development and implantation in pigs.

猪胎儿产前死亡仍是全球养猪业面临的重大关切问题。受精后至足月前，自发性胎儿丢失率可达20%至45%，且多数丢失事件发生于胚胎植入阶段。由于目前对胚胎植入阶段早期孕体（conceptus）高死亡率的调控机制尚不清楚，本研究旨在分析该阶段的整体基因表达变化，明确参与孕体发育的分子机制。本研究采用因美纳（Illumina）下一代测序技术（RNA-Seq）与实时定量PCR，对差异表达基因（differentially expressed genes, DEGs）进行分析；随后采用软聚类算法开展基因表达聚类分析。本研究在妊娠第9、12、15天的猪孕体中鉴定出8236个差异表达基因。对这些基因的注释分析显示，妊娠第9、12、15天的显著富集生物过程分别为核糖体RNA加工（GO:0006364）、细胞黏附（GO:1904874）以及心脏发育（GO:0007507）。此外，本研究还发现多个基因在猪孕体形态发育与胚胎植入过程中发挥关键作用，包括T复合体1（T-complex 1）、RuvB样AAA ATP酶2（RuvB-like AAA ATPase 2）、结缔组织生长因子、整合素、γ干扰素（interferon gamma）、SLA-1、趋化因子配体9（chemokine ligand 9）、PAG-2、转化生长因子β受体1（transforming growth factor beta receptor 1）以及膜联蛋白A2（Annexin A2）。此外，本研究还通过体外实验探究了PAG-2的功能，发现其可抑制滋养层细胞的增殖与迁移。本研究的分析结果为解析猪孕体发育与胚胎植入的调控机制提供了宝贵的研究资源。