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三种基因型猪产前骨骼肌发育的miRNA与mRNA配对表达谱的综合分析

Integrated analysis of miRNA and mRNA paired expression profiling of prenatal skeletal muscle development in three genotype pigs.

作者信息

Tang Zhonglin, Yang Yalan, Wang Zishuai, Zhao Shuanping, Mu Yulian, Li Kui

机构信息

The State Key Laboratory for Animal Nutrition, Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing 100193, China.

Agricultural Genome Institute at Shenzhen, Chinese Academy of Agricultural Sciences, Shenzhen, 518124, China.

出版信息

Sci Rep. 2015 Oct 26;5:15544. doi: 10.1038/srep15544.

Abstract

MicroRNAs (miRNAs) play a vital role in muscle development by binding to messenger RNAs (mRNAs). Based on prenatal skeletal muscle at 33, 65 and 90 days post-coitus (dpc) from Landrace, Tongcheng and Wuzhishan pigs, we carried out integrated analysis of miRNA and mRNA expression profiling. We identified 33, 18 and 67 differentially expressed miRNAs and 290, 91 and 502 mRNA targets in Landrace, Tongcheng and Wuzhishan pigs, respectively. Subsequently, 12 mRNAs and 3 miRNAs differentially expressed were validated using quantitative real-time PCR (qPCR), and 5 predicted miRNA targets were confirmed via dual luciferase reporter or western blot assays. We identified a set of miRNAs and mRNA genes differentially expressed in muscle development. Gene ontology (GO) enrichment analysis suggests that the miRNA targets are primarily involved in muscle contraction, muscle development and negative regulation of cell proliferation. Our data indicated that more mRNAs are regulated by miRNAs at earlier stages than at later stages of muscle development. Landrace and Tongcheng pigs also had longer phases of myoblast proliferation than Wuzhishan pigs. This study will be helpful to further explore miRNA-mRNA interactions in myogenesis and aid to uncover the molecular mechanisms of muscle development and phenotype variance in pigs.

摘要

微小RNA(miRNA)通过与信使RNA(mRNA)结合在肌肉发育中发挥至关重要的作用。基于长白猪、通城猪和五指山猪交配后33、65和90天的产前骨骼肌,我们对miRNA和mRNA表达谱进行了综合分析。我们分别在长白猪、通城猪和五指山猪中鉴定出33、18和67个差异表达的miRNA以及290、91和502个mRNA靶标。随后,使用定量实时PCR(qPCR)验证了12个差异表达的mRNA和3个miRNA,并通过双荧光素酶报告基因或蛋白质免疫印迹分析确认了5个预测的miRNA靶标。我们鉴定出一组在肌肉发育中差异表达的miRNA和mRNA基因。基因本体(GO)富集分析表明,miRNA靶标主要参与肌肉收缩、肌肉发育和细胞增殖的负调控。我们的数据表明,在肌肉发育的早期阶段,受miRNA调控的mRNA比后期更多。长白猪和通城猪的成肌细胞增殖阶段也比五指山猪更长。本研究将有助于进一步探索成肌过程中miRNA-mRNA的相互作用,并有助于揭示猪肌肉发育和表型变异的分子机制。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d74e/4620456/10d085816415/srep15544-f1.jpg

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