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Abstract
Non-coding RNAs especially miRNAs have been found to play important roles during skeletal muscle development. Our previous RNA-Seq performed on breast muscle tissue from 7 weeks old Recessive White Rock and Xinhua Chicken and leg muscle tissue from female Xinghua Chicken at three development time points (11 embryo age, 16 embryo age, and 1 day post hatch) (accession number GSE62971 and GSE89355, respectively) showed that miR-205a and CDH11 were differentially expressed genes. In this study, we found that overexpression of CDH11 significantly facilitated Quail muscle clone (QM7) and chicken primary myoblast (CPM) proliferation and hampered CPM differentiation. MiR-205a can directly binding to the 3′UTR of CDH11 and the overexpression of miR-205a could inhibit both cell lines (QM7) and CPM proliferation, at the meantime promote the differentiation of myoblasts. The Dual-Luciferase Reporter Assay results and qRT-PCR results showed that myogenin (MyoG) could regulate the expression of miR-205a by binding to the active region of miR-205a. Altogether our data suggest that MyoG could stimulate miR-205a expression to suppress CDH11, which promotes myoblasts proliferation while represses the differentiation.
Highlights
MicroRNAs are small non-coding RNA of 20–25 nucleotides that widely participate in chicken embryo growth and skeletal muscle development (Darnell et al, 2006; van Rooij et al, 2008; Kang et al, 2013)
MiR-205a showed a high-level expression in endoderm and ectoderm during chick embryo development (Darnell et al, 2006), so we wonder its function and mechanism in muscle development combined with our previous RNA sequencing results
Quail muscle clone 7 or chicken primary myoblast (CPM) cells cultured in 24-well plates were incubated with 50 μM 5-ethynyl-2 -deoxyuridine (EdU; RiboBio) for 2 h at 37◦C after 48 h transfection
Summary
MicroRNAs (miRNAs) are small non-coding RNA of 20–25 nucleotides that widely participate in chicken embryo growth and skeletal muscle development (Darnell et al, 2006; van Rooij et al, 2008; Kang et al, 2013). We have used high-throughput RNA sequencing to study breast muscle transcriptome in Recessive White Rock (fast-growing chicken) and Xinghua chicken (slow-growing chicken) Another high-throughput RNA sequencing was performed in leg muscles of female Xinghua chickens at 11 embryo age (E11), 16 embryo age (E16) and 1 day post hatch (D1). These databases of RNA-Seq were aimed to find miRNAs involved in skeletal muscle development at embryonic stage. Both results pointed out that miR-205a and CDH11 could be used as candidate genes associating with broiler growth (Ouyang et al, 2015; Jebessa et al, 2018). MiR-205a showed a high-level expression in endoderm and ectoderm during chick embryo development (Darnell et al, 2006), so we wonder its function and mechanism in muscle development combined with our previous RNA sequencing results
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