牛胚胎发育相关子宫液外泌体miRNA的分析和筛选OA
Analysis and Screening of miRNAs in Uterine Fluid Exosomes Related to Bovine Embryo Development
本研究通过分析超数排卵后牛子宫液外泌体中的miRNA表达差异,旨在探究其在早期妊娠中的作用,筛选与牛胚胎发育和着床相关的miRNA.本试验以皖东青年母牛为研究对象,开展了同期发情、超数排卵及人工授精.人工授精后第7天,冲洗子宫以获取胚胎及冲洗液.以国际胚胎生产平均水平5.5枚为标准,选取可用胚胎数量大于5.5枚的3头牛作为试验组,无可用胚胎的3头牛作为对照组.通过超速离心法从子宫冲洗液中提取外泌体,并使用透射电镜进行鉴定.提取外泌体中的miRNA并检测其表达水平,分析具有显著差异的miRNA.通过靶基因预测和分析,筛选和鉴定与胚胎发育和着床相关的miRNA.透射电镜结果显示,被检测样本呈茶托型,符合外泌体特征.与对照组相比,试验组中有5个miRNA显著上调、10个显著下调(P<0.05).差异最显著的已知miRNA为bta-miR-1(P=0.001).从15个差异表达的miRNA预测出2903个无重复的靶基因.KEGG分析结果表明,这些靶基因主要富集在MAPK、Rap1和黏着斑(Focal adhesion)等通路上,提示子宫液外泌体miRNA可能调控胚胎发育和着床.在牛子宫内膜上皮细胞中,bta-miR-1的过表达抑制了IGF1基因mRNA和蛋白质水平的表达,表明bta-miR-1与IGF1基因可能存在靶向关系.本研究结果可为筛选与胚胎发育和着床相关的子宫液外泌体miRNA提供参考,并为解析其在早期妊娠中的调控机制奠定基础.
By analyzing the differences of miRNA expression in uterine fluid exosomes after superovulation in cow,this study aimed to explore their roles in early pregnancy and screen for miRNAs related to bovine embryo develop-ment and implantation.The research was conducted on Wandong young female cows,involving synchronized estrus,superovulation,and artificial insemination.On the seventh day after artificial insemination,the uterus was flushed to obtain embryos and flushing fluid.Three cows with more than 5.5 available embryos were selected as the experimental group,and three cows with no available embryos were chosen as the control group,based on the international average level of 5.5 embryos.Exosomes were extracted from the uterine flushing fluid using ultracentrifugation and identified by transmission electron microscopy.miRNAs in the exosomes were extracted and their expression levels were de-tected,and miRNAs with significant differences were analyzed.Through target gene prediction and analysis,miRNAs related to embryo development and implantation were screened and identified.The transmission electron microscopy results showed that the detected samples were cup-shaped,consistent with the characteristics of exosomes.Compared with the control group,five miRNAs were significantly upregulated and ten were significantly downregulated in the ex-perimental group(P<0.05).The most significantly different known miRNA was bta-miR-1(P=0.001).The 15 differen-tially expressed miRNAs were predicted 2 903 non-repetitive target genes.KEGG analysis results indicated that these target genes were mainly enriched in the MAPK,Rap1 and focal adhesion pathways,suggesting that uterine fluid exo-some miRNAs may regulate embryo development and implantation.Overexpression of bta-miR-1 inhibited the mRNA and protein levels of the IGF1 gene,indicating a possible targeting relationship between bta-miR-1 and the IGF1 gene.The research results can provide a reference for screening uterine fluid exosome miRNAs related to embryo development and implantation and lay a foundation for understanding their regulatory mechanisms in early pregnancy.
栾曼茹;吕世杰;朱肖亭;闵佳;张子敬;施巧婷;鲁立刚;刘洪瑜;闫祥洲;王二耀
河南省农业科学院畜牧研究所,郑州 450002||河南农业大学动物科技学院,郑州 450002河南省农业科学院畜牧研究所,郑州 450002||神农种业实验室,郑州 450002河南省农业科学院畜牧研究所,郑州 450002河南省农业科学院畜牧研究所,郑州 450002||河南农业大学动物科技学院,郑州 450002河南省农业科学院畜牧研究所,郑州 450002河南省农业科学院畜牧研究所,郑州 450002毕节市农业科学院,毕节 551700安徽农业大学动物科技学院,合肥 230036河南省农业科学院畜牧研究所,郑州 450002河南省农业科学院畜牧研究所,郑州 450002
农业科技
牛外泌体miRNA差异表达妊娠
cattleexosomemiRNAdifferential expressionpregnancy
《中国草食动物科学》 2026 (2)
1-9,9
国家肉牛牦牛产业技术体系(CARS-37)河南省肉牛产业技术体系项目(HARS-22-13-S)河南省科技攻关(252102110073)河南省自然基金项目(252300420687)河南省农业科学院自主创新项目(2025ZC66)河南省农业科学院自主创新项目(2025ZC67)河南省农业科学院科技创新团队项目(2024TD25)河南省牛产业科技特派员服务团(泌阳)神农种业实验室协同创新项目(SN02-2025-02)
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