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Skeletal Muscle Transcriptome Analysis of Hanzhong Ma Duck at Different Growth Stages Using RNA-Seq

机译:使用RNA-SEQ在不同生长阶段的汉语马鸭骨骼肌转录组分析

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摘要

As one of the most important poultry worldwide, ducks (Anas platyrhynchos) are raised mainly for meat and egg products, and muscle development in ducks is important for meat production. Therefore, an investigation of gene expression in duck skeletal muscle would significantly contribute to our understanding of muscle development. In this study, twenty-four cDNA libraries were constructed from breast and leg muscles of Hanzhong Ma ducks at day 17, 21, 27 of the embryo and postnatal at 6-month-old. High-throughput sequencing and bioinformatics were used to determine the abundances and characteristics of transcripts. A total of 632,172,628 (average 52,681,052) and 637,213,938 (average 53,101,162) reads were obtained from the sequencing data of breast and leg muscles, respectively. Over 71.63% and 77.36% of the reads could be mapped to the Anas platyrhynchos genome. In the skeletal muscle of Hanzhong duck, intron variant (INTRON), synonymous variant (SYNONYMOUS_CODING), and prime 3′ UTR variant (UTR_3_PRIME) were the main single nucleotide polymorphisms (SNP) annotation information, and “INTRON”, “UTR_3_PRIME”, and downstream-gene variant (DOWNSTREAM) were the main insertion-deletion (InDel) annotation information. The predicted number of alternative splicing (AS) in all samples were mainly alternative 5′ first exon (transcription start site)-the first exon splicing (TSS) and alternative 3′ last exon (transcription terminal site)-the last exon splicing (TTS). Besides, there were 292 to 2801 annotated differentially expressed genes (DEGs) in breast muscle and 304 to 1950 annotated DEGs in leg muscle from different databases. It is worth noting that 75 DEGs in breast muscle and 49 DEGs in leg muscle were co-expressed at all developmental points of comparison, respectively. The RNA-Seq data were confirmed to be reliable by qPCR. The identified DEGs, such as CREBL2, RHEB, GDF6, SHISA2, MYLK2, ACTN3, RYR3, and STMN1, were specially highlighted, indicating their strong associations with muscle development in the Hanzhong Ma duck. KEGG pathway analysis suggested that regulation of actin cytoskeleton, oxidative phosphorylation, and focal adhesion were involved in the development of skeletal muscle. The findings from this study can contribute to future investigations of the growth and development mechanism in duck skeletal muscle.
机译:作为全球最重要的家禽之一,鸭子(Anas platyrhynchos)主要用于肉类和蛋制品,鸭子的肌肉发育对于肉类生产很重要。因此,鸭骨骼肌中基因表达的调查将显着促进我们对肌肉发育的理解。在这项研究中,在胚胎17,21,27的胚胎和6个月大的第17期,21,27天,25天的乳房和腿部肌肉构建了二十四个cDNA文库。使用高通量测序和生物信息学用于确定转录物的丰度和特征。总共632,172,628(平均52,681,052)和637,213,938(平均53,101,162)读数分别从乳房和腿部肌肉的测序数据中获得。超过71.63%和77.36%的读数可以映射到Anas platyrhynchos基因组。在汉中鸭的骨骼肌中,内含子变异(内含子),同义变体(同步)和Prime 3'UTR变体(UTR_3_prime)是主要单核苷酸多态性(SNP)注释信息,以及“Intron”,“UTR_3_PRIME”,下游 - 基因变体(下游)是主要插入删除(Indel)注释信息。所有样品中的预测数量的替代剪接(AS)主要是替代的5'第一个外显子(转录起始位点) - 第一外显子剪接(TSS)和替代3'最后的外显子(转录终端位点) - 最后的外显子剪接(TTS )。此外,乳腺肌肉中有292至2801个注释的差异表达基因(DEGS),304至1950年,来自不同数据库的腿部肌肉中的304至1950次注释。值得注意的是,在所有发育比较点分别为75次乳房肌肉和49次肌肉中的49次。 RNA-SEQ数据被QPCR确认可靠。特别突出显示所鉴定的DEG,例如Crebl2,RHEB,GDF6,Shisa2,MyLK2,Actn3,Ryr3和STMN1,表明它们在汉中马鸭中具有肌肉发育的强烈关联。 Kegg途径分析表明,肌动蛋白细胞骨架,氧化磷酸化和局灶性粘附的调节涉及骨骼肌的发育。本研究的调查结果可以有助于未来对鸭骨骼肌生长和发育机制的研究。

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