Abstract
Simple SummarySkeletal muscle is an important tissue and its development is strictly regulated by genes. In this study, in order to understand the muscle-related gene expression in Pekin duck, RNA-seq was performed to analyze and compare skeletal muscle at different growth stages. Alternative splicing, single nucleotide polymorphisms and insertion–deletions were detected, and 299 novel genes were discovered. MYL4, IGF2BP1, CSRP3, SPP1, KLHL31, LAMB2, LAMA2, ITGB1 and OPN played crucial roles in skeletal muscle development. Oxidative phosphorylation, ECM-receptor interaction, focal adhesion, carbon metabolism, and biosynthesis of amino acids participated in the regulation of skeletal muscle development in Pekin duck. This study provides an important reference for revealing the developmental mechanisms of pectoral and leg muscles in duck.Skeletal muscle, accounting for approximately 50% of body weight, is the largest and most important tissue. In this study, the gene expression profiles and pathways in skeletal muscle of Pekin duck were investigated and compared at embryonic day 17, 21, and 27 and postnatally at 6 months of age. An average of 49,555,936 reads in each sample was obtained from the transcriptome libraries. Over 70.0% of alternative splicing (AS) in each sample was 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), indicating that TSS and TTS were the most common AS event in Pekin ducks, and these AS events were closely related to the regulation of muscle development at different growth stages. The results provided a valuable genomic resource for selective breeding and functional studies of genes. A total of 299 novel genes with ≥2 exons were obtained. There were 294 to 2806 differentially expressed genes (DEGs) in each pairwise comparison of Pekin duck. Notably, 90 DEGs in breast muscle and 9 DEGs in leg muscle were co-expressed at all developmental points. DEGs were validated by qPCR analysis, which confirmed the tendency of the expression. DEGs related to muscle development were involved in biological processes such as “endodermal cell differentiation”, “muscle cell cellular homeostasis”, “skeletal muscle tissue growth” and “skeletal muscle cell differentiation”, and were involved in pathways such as oxidative phosphorylation, ECM-receptor (extracellular matrix receptor) interaction, focal adhesion, carbon metabolism, and biosynthesis of amino acids. Some DEGs, including MYL4, IGF2BP1, CSRP3, SPP1 and KLHL31, as well as LAMB2, LAMA2, ITGB1 and OPN, played crucial roles in muscle growth and development. This study provides valuable information about the expression profile of mRNAs and pathways from duck skeletal muscle at different growth stages, and further functional study of these mRNAs and pathways could provide new ideas for studying the molecular networks of growth and development in duck skeletal muscle.
Highlights
Skeletal muscle, which accounts for about 40%−50% of the bodyweight, is an important moving, defensive and metabolic organ [1]
In order to determine the potential candidate genes that may affect the development of skeletal muscle in Pekin duck, RNA-seq was used to investigate the gene expression profiles in breast and leg muscle at different growth stages
21,134,566 clean reads were received in each sample, GC content was greater than 50.37%, and Q30 value was higher than 92.59%, indicating that the sequencing data had a good reproducibility. (Table 2)
Summary
Skeletal muscle, which accounts for about 40%−50% of the bodyweight, is an important moving, defensive and metabolic organ [1]. Many studies have shown that the development of skeletal muscle is influenced by nutrition and environment, and strictly regulated by many genes, non-coding RNA and transcription factors [2,3]. Skeletal muscles grow and develop rapidly during the embryonic period, and the numbers of muscle fiber continue to increase. The postnatal muscle fiber of birds becomes hypertrophy, and the nucleus of myofiber accumulate constantly, but the numbers no longer change [4,5]. Some studies have found that Gaoyou duck and Jinding duck have different daily gains in breast and leg muscle during the incubation period [6]
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