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Deep Sequencing Analysis of miRNA Expression in Breast Muscle of Fast-Growing and Slow-Growing Broilers

机译:快速增长和缓慢增长的肉鸡胸肌中miRNA表达的深度测序分析

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摘要

Growth performance is an important economic trait in chicken. MicroRNAs (miRNAs) have been shown to play important roles in various biological processes, but their functions in chicken growth are not yet clear. To investigate the function of miRNAs in chicken growth, breast muscle tissues of the two-tail samples (highest and lowest body weight) from Recessive White Rock (WRR) and Xinghua Chickens (XH) were performed on high throughput small RNA deep sequencing. In this study, a total of 921 miRNAs were identified, including 733 known mature miRNAs and 188 novel miRNAs. There were 200, 279, 257 and 297 differentially expressed miRNAs in the comparisons of WRRh vs. WRRl, WRRh vs. XHh, WRRl vs. XHl, and XHh vs. XHl group, respectively. A total of 22 highly differentially expressed miRNAs (fold change > 2 or < 0.5; p-value < 0.05; q-value < 0.01), which also have abundant expression (read counts > 1000) were found in our comparisons. As far as two analyses (WRRh vs. WRRl, and XHh vs. XHl) are concerned, we found 80 common differentially expressed miRNAs, while 110 miRNAs were found in WRRh vs. XHh and WRRl vs. XHl. Furthermore, 26 common miRNAs were identified among all four comparisons. Four differentially expressed miRNAs (miR-223, miR-16, miR-205a and miR-222b-5p) were validated by quantitative real-time RT-PCR (qRT-PCR). Regulatory networks of interactions among miRNAs and their targets were constructed using integrative miRNA target-prediction and network-analysis. Growth hormone receptor (GHR) was confirmed as a target of miR-146b-3p by dual-luciferase assay and qPCR, indicating that miR-34c, miR-223, miR-146b-3p, miR-21 and miR-205a are key growth-related target genes in the network. These miRNAs are proposed as candidate miRNAs for future studies concerning miRNA-target function on regulation of chicken growth.
机译:生长性能是鸡的重要经济性状。 MicroRNA(miRNA)已显示在各种生物学过程中起重要作用,但它们在鸡生长中的功能尚不清楚。为了研究miRNA在鸡生长中的功能,对隐性白石(WRR)和兴化鸡(XH)的两尾标本(最高和最低体重)的胸肌组织进行了高通量小RNA深度测序。在这项研究中,总共鉴定了921个miRNA,包括733个已知的成熟miRNA和188个新颖的miRNA。在WRRh对WRR1,WRRh对XHh,WRR1对XH1和XHh对XH1组的比较中,分别有200、279、257和297个差异表达的miRNA。在我们的比较中发现总共22种高度差异表达的miRNA(倍数变化> 2或<0.5; p值<0.05; q值<0.01),它们也具有丰富的表达(读取计数> 1000)。就两个分析(WRRh与WRR1和XHh与XH1)而言,我们发现了80种常见的差异表达miRNA,而在WRRh与XHh和WRR1与XH1中发现了110个miRNA。此外,在所有四个比较中鉴定出26种常见的miRNA。通过定量实时RT-PCR(qRT-PCR)验证了四个差异表达的miRNA(miR-223,miR-16,miR-205a和miR-222b-5p)。使用整合的miRNA目标预测和网络分析构建了miRNA及其目标之间相互作用的调控网络。通过双重荧光素酶测定和qPCR证实了生长激素受体(GHR)是miR-146b-3p的靶标,表明miR-34c,miR-223,miR-146b-3p,miR-21和miR-205a是关键网络中与生长相关的靶基因。这些miRNA被提议作为候选miRNA,用于未来miRNA对鸡生长调控功能的研究。

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