College of Animal Science and Technology, Southwest University, Chongqing Key Laboratory of Herbivore Science, Chongqing Engineering Research Center for Herbivores Resource Protection and Utilization, Chongqing 400715, PR China.
College of Animal Science and Technology, Southwest University, Chongqing Key Laboratory of Herbivore Science, Chongqing Engineering Research Center for Herbivores Resource Protection and Utilization, Chongqing 400715, PR China.
Gene. 2021 Jul 20;790:145706. doi: 10.1016/j.gene.2021.145706. Epub 2021 May 9.
Previous studies have shown that extracellular vesicles (EVs) containing proteins, lipids, nucleic acids and other biological components exist in all kinds of body fluids. EVs, as an intercellular communication carrier, regulate the functions of its target cells by transporting biomacromolecules between cells. In this study, a total of six female Dazu black goats were divided into NP group (NP, non-pregnant group) and P30 (P30, 30-day pregnant group). The goats in NP group (n = 3) were in estrus, but failed to fertilize; the other goats in P30 group (n = 3) were fertilized by natural mating. Firstly, goats plasma-derived EVs were isolated using ultracentrifugation. Secondly, EVs were identified by transmission electron microscope (TEM), dynamic light scattering (DLS), and by testing its markers (CD9 and CD63) using west blotting in NP and P30 groups, respectively. Thirdly, EVs related miRNAs were sequenced and analyzed by bioinformatics method. Data shows that miR-31-5p, miR-137-3p, novel_miR_1355, novel_miR_734 and novel_miR_736 exclusively were expressed in P30 group. Their target genes were significantly enriched in the axon guidance, the Notch signaling pathway, the Wnt signaling pathway, tight junction and the Hippo signaling pathway. And miRNA-mRNA interactive network analysis reveals potential regulatory functions of miRNAs for goat during early pregnancy. These findings provided theretical references for studying the regulation of plasma-derived EVs between the fetal and placental development, and these candidate miRNAs identified might be as markers for diagnosis of goat early pregnancy.
先前的研究表明，含有蛋白质、脂质、核酸和其他生物成分的细胞外囊泡 (EVs) 存在于各种体液中。EVs 作为细胞间通讯的载体，通过在细胞间运输生物大分子来调节靶细胞的功能。在这项研究中，总共选择了 6 只大足黑山羊，分为 NP 组（NP，非妊娠组）和 P30 组（P30，妊娠 30 天组）。NP 组的 3 只山羊处于发情期，但未能受精；P30 组的另外 3 只山羊通过自然交配受精。首先，使用超速离心法从山羊血浆中分离 EVs。然后，通过透射电子显微镜（TEM）、动态光散射（DLS）以及分别在 NP 和 P30 组中使用 Western blot 检测其标记物（CD9 和 CD63）来鉴定 EVs。第三，通过生物信息学方法对 EVs 相关的 miRNAs 进行测序和分析。结果表明，miR-31-5p、miR-137-3p、novel_miR_1355、novel_miR_734 和 novel_miR_736 在 P30 组中特异性表达。它们的靶基因在轴突导向、Notch 信号通路、Wnt 信号通路、紧密连接和 Hippo 信号通路中显著富集。miRNA-mRNA 相互作用网络分析揭示了 miRNAs 在山羊早期妊娠期间对其的潜在调控功能。这些发现为研究胎儿和胎盘发育过程中血浆源性 EVs 的调节提供了理论参考，并且鉴定出的这些候选 miRNAs 可能作为山羊早期妊娠诊断的标志物。
