MicroRNAome of Porcine Conceptuses and Trophoblasts: Expression Profile of microRNAs and Their Potential to Regulate Genes Crucial for Establishment of Pregnancy
MicroRNAome of Porcine Conceptuses and Trophoblasts: Expression Profile of microRNAs and Their Potential to Regulate Genes Crucial for Establishment of Pregnancy
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DOI：
10.1095/biolreprod.114.123588
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发表时间：
2015-01-01
影响因子：
3.6
通讯作者：
Kaczmarek, Monika M.
中科院分区：
文献类型：
--
作者：
Krawczynski, Kamil;Najmula, Joanna;Kaczmarek, Monika M.
紧密协调、互惠的胚胎和母亲的相互作用会影响怀孕早期的基因表达。最近，microRNAs(MiRNAs)通过转录后基因调控机制在哺乳动物胚胎发育和植入的微调中发挥了新的作用。在这里，我们结合了转录和计算方法来描述猪妊娠早期胚胎和滋养层不同发育阶段的miRNAs和miRNA合成和运输相关基因。利用半定量RT-PCR技术，我们检测了与miRNA合成和转运相关的10个编码蛋白的基因的mRNA水平：DROSHA、Dgcr8、XPO5、DICER1、TARBP2、TNRC6A、AGO1、AGO2、AG03和AGO4。定制的多物种微阵列被用来分析miRNAs。MiRNA-mRNA相互作用的预测算法允许识别分析的miRNAs的目标转录本。这些包括血管内皮生长因子、激光诱导因子、Ptgs2和IL-6R，它们被认为是猪胚胎-母体相互作用的关键成分。选择了两个miRNAs，miR-26a和miR-125b，用于检测妊娠期间子宫腔冲洗分离的细胞外小泡中的存在情况。体外研究结果表明，miR-125b等miRNAs可以调节猪子宫内膜腔上皮细胞中胚胎发育和着床的关键基因的表达。首次报道了猪妊娠母体识别和胚胎着床过程中miRNAs及相关基因在猪胚胎和滋养层中的表达谱。总之，我们的结果表明，这些小的，非编码的RNA在猪早期胚胎发育和早期怀孕期间的胚胎-母体相互作用中具有潜在的作用。
Tightly coordinated, reciprocal embryo-maternal interactions affect gene expression during early pregnancy. Recently, microRNAs (miRNAs) have emerged as new players in the fine tuning of embryo development and implantation in mammals via posttranscriptional gene regulation mechanisms. Here, we integrated transcriptomic and computational approaches to profile miRNAs and miRNA synthesis and transport-related genes at different developmental stages of porcine conceptuses and trophoblast during early pregnancy in the pig. Using semiquantitative RT-PCR, we examined mRNA levels of 10 genes encoding proteins involved in miRNA synthesis and transport: DROSHA, DGCR8, XPO5, DICER1, TARBP2, TNRC6A, AGO1, AGO2, AGO3, and AGO4. Custom, multispecies microarrays were used to profile miRNAs. Prediction algorithms of miRNA-mRNA interactions allowed identification of target transcripts for the analyzed miRNAs. These included VEGF, LIF, PTGS2, and IL-6R, known to be crucial components of embryo-maternal interactions in the pig. Two selected miRNAs, miR-26a and miR-125b, were tested for the presence in the extracellular vesicles isolated from uterine luminal flushings during pregnancy. Results of in vitro study demonstrated that miRNAs, such as miR-125b, can regulate expression of genes crucial for embryo development and implantation in porcine endometrial luminal epithelial cells. For the first time, expression profiles of miRNAs and related genes in porcine conceptuses and trophoblast during maternal recognition of pregnancy and embryo implantation in the pig were described. Altogether, our results indicate potential roles of these small, noncoding RNAs in the early development of embryos and embryo-maternal cross-talk during early pregnancy in the pig.