| 胚胎移植前供体牛与受体牛血浆外泌体miRNA差异表达分析 |
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| 引用本文： | 翟亚莹,施巧婷,楚秋霞,朱肖亭,滑留帅,张子敬,陈付英,祁兴磊,王二耀,吕世杰.胚胎移植前供体牛与受体牛血浆外泌体miRNA差异表达分析[J].中国畜牧兽医,2022,49(2):641-649. |
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| 作者姓名： | 翟亚莹 施巧婷 楚秋霞 朱肖亭 滑留帅 张子敬 陈付英 祁兴磊 王二耀 吕世杰 |
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| 作者单位： | 1. 河南省农业科学院畜牧兽医研究所, 河南省畜禽繁育与营养调控重点实验室, 郑州 450002;2. 河南农业大学动物科技学院, 郑州 450008;3. 泌阳县畜牧技术服务中心, 驻马店 463700 |
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| 基金项目： | 河南省农业科学院基本科研业务费项目(2021ZC49);河南省农业科学院科技创新创意项目(2020CX09);河南省重大公益专项项目(201300111200);中原学者工作站项目(214400510013);河南省肉牛产业技术体系项目(S2013-08);国家肉牛牦牛产业技术体系项目(CARS-37) |
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| 摘 要： | 【目的】 探索胚胎移植前供体牛与受体牛血浆外泌体miRNA的表达差异, 以明确血浆外泌体miRNA在牛早期妊娠中的作用及其调控机制。【方法】 以3~6岁、体重480~600 kg的夏南牛作为研究对象, 选取10头供体牛进行同期发情、超数排卵和人工授精, 23头受体牛只做同期发情处理。在人工授精后第7天, 冲洗供体牛子宫以获取囊胚, 选取3头囊胚数相近的供体牛及3头与供体牛体重和年龄均相近的受体牛, 颈静脉采血, 进行血浆外泌体的分离与鉴定; 然后提取血浆外泌体miRNA, 并检测其表达量; 采用R语言中的DESeq差异算法计算P值, 并筛选出P<0.05的miRNA, 对差异表达的miRNA进行靶基因预测、GO功能富集分析和KEGG信号通路分析。【结果】 6个样本的囊泡粒径均在135 nm左右, 符合外泌体的特征。与供体牛相比, 受体牛中有9个miRNAs表达显著上调(P<0.05), 13个miRNAs显著下调(P<0.05);22个差异表达的miRNAs中, 有15个miRNAs预测出无重复的靶基因2 990个。GO功能富集分析和KEGG信号通路分析的结果表明, 这些靶基因主要富集在与生物黏附(biological adhesion)、定位(localization)、细胞连接(cell junction)功能有关的通路上, 显著富集的信号通路与黏着斑(focal adhesion)、黏着连接(adherens junction)有关, 提示血浆外泌体miRNA可能参与调控胚胎着床。【结论】 研究结果可为筛选和探究影响胚胎着床的血浆外泌体miRNA提供参考, 并为进一步阐明血浆外泌体miRNA在母牛早期妊娠调控中的作用提供依据。
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| 关 键 词： | 牛 外泌体 miRNA 差异表达 妊娠 |
| 收稿时间： | 2021-07-27 |
Differential Expression Analysis of Plasma Exosomes microRNA Between Donor and Recipient Cattle Before Embryo Transfer |
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| ZHAI Yaying,SHI Qiaoting,CHU Qiuxia,ZHU Xiaoting,HUA Liushuai,ZHANG Zijing,CHEN Fuying,QI Xinglei,WANG Eryao,LYU Shijie.Differential Expression Analysis of Plasma Exosomes microRNA Between Donor and Recipient Cattle Before Embryo Transfer[J].China Animal Husbandry & Veterinary Medicine,2022,49(2):641-649. |
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| Authors: | ZHAI Yaying SHI Qiaoting CHU Qiuxia ZHU Xiaoting HUA Liushuai ZHANG Zijing CHEN Fuying QI Xinglei WANG Eryao LYU Shijie |
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| Institution: | 1. Henan Key Laboratory of Livestock and Poultry Breeding and Nutrition Regulation, Institute of Animal Science and Veterinary Medicine, Henan Academy of Agricultural Sciences, Zhengzhou 450002, China;2. College of Animal Science and Technology, Henan Agricultural University, Zhengzhou 450008, China;3. Center of Animal Husbandry Technical Service in Biyang, Zhumadian 463700, China |
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| Abstract: | 【Objective】 This study was to explore the difference in the expression of plasma exosome miRNA between donor and recipient cattle before embryo transfer, and to detemine the role of plasma exosome miRNA in bovine pregnancy.【Method】 Xianan cattle aged 3-6 years and weighing 480-600 kg were used as the research object, 10 donor cattle were selected for simultaneous estrus, superovulation and artificial insemination, and 23 recipient cattle were treated for simultaneous estrus. On the 7th day after artificial insemination, the donor bovine uterus was washed to obtain blastocysts, 3 donor cattle with similar number of blastocysts and three recipient cattle with similar body weight and age to the donor cattle were selected to collect blood from jugular vein, and plasma exosomes were collected.After extracting miRNA from plasma exosomes, the expression level was analyzed.Target genes of differentially expressed miRNA were predicted.While the P value was calculated with DESeq algorithm in R package, the miRNA with P < 0.05 were screened, and the differentially expressed miRNA were analyzed by target gene prediction, GO function enrichment analysis and KEGG signaling pathway analysis.【Result】 The vesicles particle size of the 6 samples was about 135 nm, which was consistent with the diameter characteristics of exosomes.Nine miRNAs were significantly up regulated in recipient cattle compared with donor cattle (P < 0.05), and 13 miRNAs were significantly down regulated (P < 0.05).Among the 22 differentially expressed miRNAs, 15 miRNAs predicted 2 990 target genes without duplication.The results of GO function enrichment analysis and KEGG signal pathway analysis indicated that these target genes were mainly concentrated in the functional pathways related to biological adhesion, localization and cell junction, and significant accumulation of signal pathway was related to focal adhesion and adherens junction, suggested that plasma exosomal miRNA might be involved in the regulation of embryo implantation.【Conclusion】 These results could provide reference for screening and exploring the plasma exosomal miRNA affecting embryo implantation, and provide a basis for further elucidation of the role of plasma exosomal miRNA in the regulation of early pregnancy in cattle. |
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| Keywords: | cattle exosome miRNA differential expression pregnancy |
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