巴什拜羊骨骼肌不同发育阶段差异表达miRNA研究  

Research of Differentially Expressed miRNA of Skeletal Muscle During Different Development Stages in Bashbay Sheep(Ovis aries)

作　　者：张伟[1,2] 王世银[1] 邓双义[1] 杨力伟[1] 石国庆[2] 甘尚权[2] 

机构地区：[1]新疆农业职业技术学院,昌吉831100 [2]新疆农垦科学院畜牧兽医研究所省部共建绵羊遗传改良与健康养殖重点实验室,石河子832000

出　　处：《农业生物技术学报》2018年第1期104-112,共9页Journal of Agricultural Biotechnology

基　　金：新疆维吾尔自治区高等学校科学研究计划重点项目(No.XJEDU2016I062);新疆农业职业技术学院科研项目(No.XJNZYKJ2016017)

摘　　要：miRNA在绵羊(Ovis aries)骨骼肌生长发育过程中发挥着重要的调控作用。为了挖掘巴什拜羊骨骼肌不同发育阶段差异表达miRNA,为深入解析miRNA与巴什拜羊优良肉质性状的关联性提供基础数据,本研究应用Solexa测序技术对巴什拜羊胎儿期(40,50,60,80,100和120 d)及出生当天和出生后(30,60和90 d)骨骼肌组织混合样品的miRNA表达情况及差异表达miRNA进行了研究。胎儿期和出生后骨骼肌组织中分别测得11.82 M和11.51 M的clean reads,及0.31和0.28 M的unique sequences,其中miRNA序列分别占Unique sequences的20%和17%。通过与miRBase(V21.0)收录的miRNA序列比对,胎儿期和出生后分别获得537个和496个物种间保守miRNA,分别有102个和115个与绵羊miRNA相匹配。剩余序列比对至绵羊表达序列标签(expressed sequence tags,ESTs)数据库,结合Mfold和Mi Pred程序预测miRNA,分别预测到486个和510个新的miRNA。胎儿期和出生后共得到385个差异表达的miRNA(表达量变化2倍以上),其中上调203个,下调182个。采用Target Scan、Pictar和miRanda软件对上调和下调前10位共计20个miRNA的靶基因进行预测,并选择至少出现在两个软件预测结果中的靶基因参与后续分析,共预测3 852个靶基因。进一步分析表明,预测到的靶基因参与促分裂素原活化蛋白激酶(mitogen-activated protein kinases,MAPK)、WNT(wingless type MMTV integration site family members)和黏附连接(adherens junction)等重要的信号通路。本研究结果丰富了绵羊骨骼肌中miRNA表达的相关数据,为进一步阐明miRNA在绵羊骨骼肌发育中的调控机制提供了依据。miRNA plays an important role in sheep （Ovis aries） skeletal muscle development. This study aimed to find out the differently expressed miRNA in Bashbay sheep skeletal muscle during different development stages, and to furtherly understand the relationship between miRNA expression and excellent meat traits of Bashbay sheep. In this study, the Solexa sequencing technology was applied to investigate the miRNA expressing profiles in Bashbay sheep skeletal muscle during fetal period （mixed sample of 40, 50, 60, 80, 100 and 120 d） and postnatal stage （mixed sample of newborn, 30, 60 and 90 d）. By high-throughput sequencing and data analysis, 11.82 M and 11.51 M clean reads, and 0.31 M 0.28 M unique sequences were obtained. Among them, 20% and 17% were candidate miRNA sequences, respectively. Then blastn was performed between miRBase （V21.0） and unique sequences, and finally got 537 and 496 miRNAs which were conservative among species. There were 102 and 115 miRNAs of them were matched with ovine known miRNAs, respectively. The rest sequences were compared with EST database of sheep, and 486 and 510 new candidate miRNAs were obtained by Mfold and MiPred program. There were total 385 miRNAs which differently expressed between fetal and postnatal skeletal muscle of Bashbay sheep （expressing change were above 2 fold）, including 203 up-regulated and 182 down-regulated miRNAs, respectively. Applying TargetScan, Pictar and miRanda, the candidate target genes of top 10 up- and down-regulated miRNAs were predicted, and the one which was predicted by at least two programs was chosen to be further analyzed. Total 3 852 candidate target genes were predicted, and most of them participated in many important signal pathways, just like mitogen-activated protein kinases （MAPK）, wingless type MMTV （mouse mammary tumor virus） integration site family members （WNT） and adherens junction, etc. This study enriches the miRNA database of sheep skeletal muscle, and could help to further understand the regulation

关 键 词：巴什拜羊 骨骼肌 MIRNA 差异表达 发育 

分 类 号：S826.8[农业科学—畜牧学]