乌头驴与三粉驴骨骼肌中miRNAs的表达鉴定与分析

doi:10.16431/j.cnki.1671-7236.2021.06.002

Abstract

Abstract: To investigate the expression of microRNAs(miRNAs) in skeletal muscles of Dezhou donkeys and further improve the regulatory mechanism of small RNA in the development of skeletal muscle of donkey.In this study,the longissimus dorsi muscle of two Dezhou donkey strains (Wutou donkey and Sanfen donkey) was selected as the research objects,by constructing small RNA sequencing library of Wutou donkey longissimus dorsi muscle (WB) and Sanfen donkey longissimus dorsi muscle (SB),bioinformatics techniques were used to carry out differential expression analysis of miRNAs between WB and SB,and predict and analyze miRNAs that might affect the meat production traits of donkey.In addition,six miRNAs with high expression were selected,and Real-time quantitative PCR was used to verify the accuracy of sequencing results.The results showed that 982 and 1 149 conserved miRNAs and 106 and 143 new miRNAs were identified in WB and SB libraries.In WB and SB comparison group (WB vs SB) based on conditions|log₂(fold_change)|≥2,P≤0.05 get 17 differentially expressed miRNAs,of which 5 were up-regulated and 12 were down-regulated.Through GO annotation,it found that differentially expressed miRNAs were significantly enriched in biological processes such as regulating cell migration and differentiation,cell division and skeletal muscle cell contraction (P<0.05).bta-miR-378d_L+1R-1_1ss22CA and bta-miR-378d_R-2 which related to skeletal muscle development in donkey was found,they target multiple genes such as NPY1R,GPR152,BEST1,KCNH8 and SPAG9.KEGG enrichment results showed that miRNAs were enriched in Wnt,MAPK,ubiquitin proteolysis system and other signaling pathways related to skeletal muscle development.Among them,a total of 207 target genes 17 differentially expressed miRNAs were involved in the Wnt signaling pathway,especially PC-5p-84483_31,hsa-miR-186-3p_R-3 and cfa-miR-329b_1ss19CT had the largest number of target genes.The results of Real-time quantitative PCR were consistent with the results of miRNAs sequencing,and it found that the expression levels of the other five miRNAs were all higher in the SB than in the WB except for eca-miR-181b.In this study,miRNAs of skeletal muscle of two strains of Dezhou donkey were analyzed for the first time,and the differences of RNA-seq between Wutou donkey and Sanfen donkey were revealed,which provided reference for elucidating the molecular regulation mechanism of skeletal muscle growth and development of Dezhou donkey.

Key words: Dezhou donkey; skeletal muscle development; transcriptome; miRNA

CLC Number:

S822

MA Cheng, HU Wenping, SHI Tianpei, HOU Haobin, LI Haijing, ZHANG Xinhao, YANG Li, ZHANG Li. Identification and Analysis of miRNAs Expression in Skeletal Muscle of Wutou Donkey and Sanfen Donkey[J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(6): 1894-1905.

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Identification and Analysis of miRNAs Expression in Skeletal Muscle of Wutou Donkey and Sanfen Donkey

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