Hu Zhigang, Cao Junting, Zhang Jianqin, Ge Liyan, Zhang Huilin, Liu Xiaolin
College of Animal Science and Technology, Northwest A&F University, Yangling 712100, Shaanxi, China.
Biomolecules. 2021 Feb 19;11(2):315. doi: 10.3390/biom11020315.
As one of the most important poultry worldwide, ducks () 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 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 2, , 6, 2, 2, 3, 3, and 1, 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.
作为全球最重要的家禽之一,鸭主要用于生产肉和蛋产品,鸭的肌肉发育对肉类生产很重要。因此,研究鸭骨骼肌中的基因表达将极大地有助于我们了解肌肉发育。在本研究中,从汉中麻鸭胚胎期第17、21、27天以及出生后6个月大的胸肌和腿肌构建了24个cDNA文库。利用高通量测序和生物信息学来确定转录本的丰度和特征。分别从胸肌和腿肌的测序数据中获得了总计632,172,628条(平均52,681,052条)和637,213,938条(平均53,101,162条) reads。超过71.63%和77.36%的reads能够映射到基因组。在汉中鸭的骨骼肌中,内含子变异(INTRON)、同义变异(SYNONYMOUS_CODING)和3'UTR末端变异(UTR_3_PRIME)是主要的单核苷酸多态性(SNP)注释信息,而“INTRON”、“UTR_3_PRIME”和下游基因变异(DOWNSTREAM)是主要的插入缺失(InDel)注释信息。所有样本中预测的可变剪接(AS)数量主要是可变5'第一外显子(转录起始位点)-第一外显子剪接(TSS)和可变3'最后外显子(转录终止位点)-最后外显子剪接(TTS)。此外,来自不同数据库的胸肌中有292至2801个注释的差异表达基因(DEG),腿肌中有304至1950个注释的DEG。值得注意的是,胸肌中的75个DEG和腿肌中的49个DEG在所有发育比较点均共表达。通过qPCR证实RNA-Seq数据可靠。所鉴定的DEG,如2、、6、2、2、3、3和1,被特别突出显示,表明它们与汉中麻鸭的肌肉发育密切相关。KEGG通路分析表明,肌动蛋白细胞骨架调节、氧化磷酸化和粘着斑参与了骨骼肌的发育。本研究结果有助于未来对鸭骨骼肌生长发育机制的研究。