Abstract
Simple SummaryIt is of great commercial interest to elucidate the genetic mechanisms associated with skeletal muscle development in the duck. In this study, we performed high throughput microRNA (miRNA) sequencing to identify the candidate miRNAs during two developmental stages of duck embryonic breast muscle. We detected 1091 miRNAs and 109 of them were differentially expressed between embryonic day 13 (E13) and E19. We also predicted the target genes of the differentially expressed miRNAs and subsequently analyzed the enriched gene ontology (GO) terms and Kyoto Encyclopedia of Genes and Genomes (KEGG) signaling pathways, and finally constructed a protein–protein interaction (PPI) network with the target genes. Luciferase reporter assay showed that the growth-related genes, Fibroblast growth factor receptor like 1 (FGFRL1) and Insulin like growth factor 2 mRNA binding protein 1 (IGF2BP1), were target genes of miR-214-5p. These results can supplement the duck miRNA database and provide several candidate miRNAs for future studies on the regulation of embryonic skeletal muscle development.Poultry skeletal muscle provides high quality protein for humans. Study of the genetic mechanisms during duck skeletal muscle development contribute to future duck breeding and meat production. In the current study, three breast muscle samples from Shan Ma ducks at embryonic day 13 (E13) and E19 were collected, respectively. We detected microRNA (miRNA) expression using high throughput sequencing following bioinformatic analysis. qRT-PCR validated the reliability of sequencing results. We also identified target prediction results using the luciferase reporter assay. A total of 812 known miRNAs and 279 novel miRNAs were detected in six samples; as a result, 61 up-regulated and 48 down-regulated differentially expressed miRNAs were identified between E13 and E19 (|log2 fold change| ≥ 1 and p ≤ 0.05). Enrichment analysis showed that target genes of the differentially expressed miRNAs were enriched on many muscle development-related gene ontology (GO) terms and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways, especially mitogen-activated protein kinase (MAPK) signaling pathways. An interaction network was constructed using the target genes of the differentially expressed miRNAs. These results complement the current duck miRNA database and offer several miRNA candidates for future studies of skeletal muscle development in the duck.
Highlights
IntroductionFood conversion ratio and meat production are important economic indicators in livestock and the growth and development of skeletal muscle has an appreciable impact on meat production
Skeletal muscle forms a large proportion of the body of animals
A total of 30 fertile Shan Ma duck eggs from the same batch were purchased from Jiangxi Tianyun duck breeding farm (Nanchang, Jiangxi, China): 15 embryos were at embryonic day 13 (E13) and 15 embryos at Embryonic day 19 (E19)
Summary
Food conversion ratio and meat production are important economic indicators in livestock and the growth and development of skeletal muscle has an appreciable impact on meat production. An exploration of the molecular mechanisms related to skeletal muscle growth and development is important to human food security. Skeletal muscle development can be divided into two stages: myoblast proliferation and the hypertrophy of myotubes. Myoblasts proliferate and differentiate into multi-nucleated myotubes; subsequently, myotubes further differentiate into mature muscle fibers [1]. There are many molecules that regulate the growth and development of poultry muscle, including protein-coding genes and non-coding RNAs [3,4,5]. Embryonic day 19 (E19) is a crucial stage of duck breast muscle development. Myogenic factor 6 (MRF4) and myogenin (MyoG) have the highest expression level and myostatin (MSTN) has the lowest expression level in this stage, the breast muscle weight is nearly constant from E19 to hatching [6]
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