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Skeletal muscle differentiation can be regulated by various transcription factors and non-coding RNAs. In our previous work, miR-223 is differentially expressed in the skeletal muscle of chicken with different growth rates, but its role, expression and action mechanism in muscle development still remains unknown. Here, we found that MYOD transcription factor can upregulate miR-223 expression by binding to an E-box region of the  gga-miR-223  gene promoter during avian myoblast differentiation.  IGF2  and  ZEB1  are two target genes of miR-223. The target inhibition of miR-223 on  IGF2  and  ZEB1  are dynamic from proliferation to differentiation of myoblast. miR-223 inhibits  IGF2  expression only in the proliferating myoblast, whereas it inhibits  ZEB1  mainly in the differentiating myoblast. The inhibition of  IGF2  by miR-223 resulted in the repression of myoblast proliferation. During myoblast differentiation, miR-223 would be upregulated owing to the promoting effect of MYOD, and the upregulation of miR-223 would inhibit  ZEB1  to promote myoblast differentiation. These results not only demonstrated that the well-known muscle determination factor MYOD can promote myoblast differentiation by upregulate miR-223 transcription, but also identified that miR-223 can influence myoblast proliferation and differentiation by a dynamic manner regulates the expression of its target genes.

miRNA-223 upregulated by MYOD inhibits myoblast proliferation by repressing IGF2 and facilitates myoblast differentiation by inhibiting ZEB1