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首页> 外文期刊>Biology of Reproduction: Offical Journal of the Society for the Study of Reproduction >Deep Sequencing of the Porcine Endometrial Transcriptome on Day 14 of Pregnancy
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Deep Sequencing of the Porcine Endometrial Transcriptome on Day 14 of Pregnancy

机译:妊娠第14天猪子宫内膜转录组的深度测序

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In pigs, conceptus attachment to the uterine surface epithelium starts around Day 14 of pregnancy preceded by a pronounced vascularization at the implantation zones, initiating the epitheliochorial placentation. To characterize the complex transcriptome changes in the endometrium in the course of initial conceptus attachment, deep sequencing of endometrial RNA samples of pregnant animals (n = 4) and corresponding cyclic controls (n = 4) was performed using Illumina RNA-Seq. The obtained sequence reads were mapped to the porcine genome, and relative expression values were calculated for the analysis of differential gene expression. Statistical analysis revealed 1933 differentially expressed genes (false discovery rate 1%), 1229 with higher and 704 with lower mRNA concentration, in the samples from pregnant animals. Expression of selected genes was validated by the use of quantitative realtime RT-PCR. The RNA-Seq data were compared to results of a microarray study of bovine endometrium on Day 18 of pregnancy and additional related data sets. Bioinformatics analysis revealed for the genes with higher mRNA concentration in pregnant samples strong overrepresentation, particularly for immune-related functional terms but also for apoptosis and cell adhesion. Overrepresented terms for the genes with lower mRNA concentration in pregnant samples were related to extracellular region, ion transport, cell adhesion, and lipid and steroid metabolic process. In conclusion, RNA-Seq analysis revealed comprehensive transcriptome differences in porcine endometrium between Day 14 of pregnancy and corresponding cyclic endometrium and highlighted new processes and pathways probably involved in regulation of noninvasive implantation in the pig.
机译:在猪中,子宫附着在子宫表面上皮开始于妊娠的第14天左右,然后在植入区出现明显的血管形成,从而开始上皮膜上的胎盘形成。为了表征初始概念附着过程中子宫内膜的复杂转录组变化,使用Illumina RNA-Seq对怀孕动物的子宫内膜RNA样品(n = 4)和相应的循环对照(n = 4)进行了深度测序。将获得的序列读数定位到猪基因组,并计算相对表达值以分析差异基因表达。统计分析显示,来自怀孕动物的样品中有1933个差异表达基因(错误发现率为1%),1229个具有较高的mRNA表达和704个具有较低的mRNA浓度。通过使用定量实时RT-PCR来验证所选基因的表达。将RNA-Seq数据与妊娠第18天牛子宫内膜的微阵列研究结果及其他相关数据集进行比较。生物信息学分析表明,对于孕妇样品中具有较高mRNA浓度的基因,存在明显的过度表达,尤其是与免疫相关的功能术语,以及细胞凋亡和细胞粘附。孕妇样品中具有较低mRNA浓度的基因的过分表述与细胞外区域,离子转运,细胞粘附以及脂质和类固醇代谢过程有关。总之,RNA-Seq分析揭示了在怀孕第14天和相应的环状子宫内膜之间猪子宫内膜的全面转录组差异,并强调了可能参与调节猪非侵入性植入的新过程和途径。

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