Abstract
A series of elaborately regulated and orchestrated changes in gene expression profiles leads to muscle growth and development. In this study, RNA sequencing was used to profile embryonic chicken myoblasts and fused myotube transcriptomes, long non-coding RNAs (lncRNAs), and messenger RNAs (mRNAs) at four stages of myoblast differentiation. Of a total of 2484 lncRNA transcripts, 2288 were long intergenic non-coding RNAs (lincRNAs) and 198 were antisense lncRNAs. Additionally, 1530 lncRNAs were neighboring 2041 protein-coding genes (<10 kb upstream and downstream) and functionally enriched in several pathways related to skeletal muscle development that have been extensively studied, indicating that these genes may be in cis-regulatory relationships. In addition, Pearson’s correlation coefficients demonstrated that 990 lncRNAs and 7436 mRNAs were possibly in trans-regulatory relationships. These co-expressed mRNAs were enriched in various developmentally-related biological processes, such as myocyte proliferation and differentiation, myoblast differentiation, and myoblast fusion. The number of transcripts (906 lncRNAs and 4422 mRNAs) differentially expressed across various stages declined with the progression of differentiation. Then, 4422 differentially expressed genes were assigned to four clusters according to K-means analysis. Genes in the K1 cluster likely play important roles in myoblast proliferation and those in the K4 cluster were likely associated with the initiation of myoblast differentiation, while genes in the K2 and K3 clusters were likely related to myoblast fusion. This study provides a catalog of chicken lncRNAs and mRNAs for further experimental investigations and facilitates a better understanding of skeletal muscle development.
Highlights
Skeletal muscle, which accounts for approximately 40% of the body weight of mammals, is an important tissue involved in the regulation of metabolism, locomotion, and strength [1]
This study provides a catalog of chicken long non-coding RNAs (lncRNAs) and messenger RNAs (mRNAs) for further experimental investigations and facilitates a better understanding of skeletal muscle development
Chicken myoblasts myoblasts were were derived derived from from primary primary cultures cultures of of embryonic embryonic breast breast muscle muscle tissues tissues and and Chicken fused myotubes were generated by culturing the myoblasts in differentiation medium for
Summary
Skeletal muscle, which accounts for approximately 40% of the body weight of mammals, is an important tissue involved in the regulation of metabolism, locomotion, and strength [1]. A loss or reduction in skeletal muscle mass results in weakness and impaired mobility and, if severe enough, in increased morbidity and mortality [2,3]. Skeletal muscle develops into meat, which is the primary terminal product for human consumption. The study of muscle development in agriculturally-important species is essential to achievements of increased body weight and muscle mass. The process of muscle fiber development is nearly completed during the prenatal stage. The development of skeletal muscle depends on myogenesis, which is a multistep process starting with myoblast proliferation, followed by their exit from cell cycle, Genes 2018, 9, 34; doi:10.3390/genes9010034 www.mdpi.com/journal/genes
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