Screening and Identification of Plasma Exosomal miRNAs in Early Pregnancy in JunKen Mutton Sheep (Ovis aries)
LYU Zhan-Min1,2, ZHANG Zhen-Liang1,2, LIU Yu-Cheng1,2, WANG Jing-Jing1,2, LIU Wei-Jun1,2, TANG Ling1,2, HUA Yu-Xuan1,2, NI Jian-Hong1,2, WAN Peng-Cheng1,2, LU Chun-Xia1,3,*, LIU Chang-Bin1,2,*
1 Institute of Animal Husbandry and Veterinary Science, Xinjiang Academy of Agriculture and Reclamation Sciences, Shihezi 832000, China; 2 State Key Laboratory of Sheep Genetic Improvement and Healthy Production, Shihezi 832000, China; 3 School of Advanced Agriculture and Bioengineering, Yangtze Normal University, Chongqing 408100, China
Abstract：Early pregnancy is a critical factor influencing sheep (Ovis aries) reproduction. Accurate identification of non-pregnant sheep can effectively reduce the empty period and enhance reproductive efficiency. Exosomes, which are extracellular vesicles with diameters ranging from 30 to 150 nm, are secreted by cells and play a crucial role in nearly all aspects of mammalian pregnancy. In this study, JunKen mutton sheep ewes aged 2 to 3 years were selected as research subjects. Plasma samples were collected from 3 ewes at non-pregnant (0 d) and early pregnant stages (14, 30 d). Plasma exosomes were isolated to construct miRNA libraries from these JunKen mutton sheep at different early pregnancy time points. Significantly differentially expressed miRNAs were identified in the plasma exosomes of JunKen mutton sheep at various early pregnancy stages. Through library analysis and database alignment, a total of 248 differentially expressed miRNAs were detected, including 79 known and 169 novel miRNAs. Comparative analysis revealed 40 differentially expressed miRNAs between 14 and 0 d (30 up-regulated, 10 down-regulated), 72 between 30 d and 0 d (43 up-regulated, 29 down-regulated), and 35 between 30 d and 14 d (9 up-regulated, 26 down-regulated). Target gene annotation and functional enrichment of differentially expressed exosomal miRNAs were performed by aligning with the GO and KEGG databases using BLAST software. Enriched biological processes were found to be associated with early pregnancy. Notably, oar-miR-27a exhibited high expression levels and significant differences between non-pregnant 0 d and pregnant 14 d and 30 d, suggesting its potential role in early pregnancy and embryonic development in sheep. To validate the sequencing results, 5 differentially expressed miRNAs were randomly selected for expression detection using qRT-PCR, and the results were consistent with the sequencing data. This study demonstrated that plasma exosomal miRNAs might participate in the regulation of early pregnancy in JunKen mutton sheep. These findings provide new insights into the regulatory processes of early pregnancy in sheep and offer a theoretical basis for exploring the molecular mechanisms by which differentially expressed exosomes influence early pregnancy in this species.
吕占民, 张振良, 刘昱成, 王晶晶, 刘伟俊, 唐玲, 华雨萱, 倪建宏, 万鹏程, 卢春霞, 刘长彬. 军垦肉羊妊娠早期血浆外泌体miRNA的筛选和鉴定[J]. 农业生物技术学报, 2025, 33(9): 1993-2006.
LYU Zhan-Min, ZHANG Zhen-Liang, LIU Yu-Cheng, WANG Jing-Jing, LIU Wei-Jun, TANG Ling, HUA Yu-Xuan, NI Jian-Hong, WAN Peng-Cheng, LU Chun-Xia, LIU Chang-Bin. Screening and Identification of Plasma Exosomal miRNAs in Early Pregnancy in JunKen Mutton Sheep (Ovis aries). 农业生物技术学报, 2025, 33(9): 1993-2006.
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