Abstract
Previous studies have shown that gga-miR-2954 was highly expressed in the gonads and other tissues of male chickens, including muscle tissue. Yin Yang1 (YY1), which has functions in mammalian skeletal muscle development, was predicted to be a target gene of gga-miR-2954. The purpose of this study was to investigate whether gga-miR-2954 plays a role in skeletal muscle development by targeting YY1, and evaluate its function in the sexual dimorphism development of chicken muscle. Here, all the temporal and spatial expression profiles in chicken embryonic muscles showed that gga-miR-2954 is highly expressed in males and mainly localized in cytoplasm. Gga-miR-2954 exhibited upregulated expression of in vitro myoblast differentiation stages. Next, through the overexpression and loss-of-function experiments performed in chicken primary myoblasts, we found that gga-miR-2954 inhibited myoblast proliferation but promoted differentiation. During myogenesis, gga-miR-2954 could suppress the expression of YY1, which promoted myoblast proliferation and inhibited the process of myoblast cell differentiation into multinucleated myotubes. Overall, these findings reveal a novel role of gga-miR-2954 in skeletal muscle development through its function of the myoblast proliferation and differentiation by suppressing the expression of YY1. Moreover, gga-miR-2954 may contribute to the sex difference in chicken muscle development.
Highlights
In vertebrates, skeletal muscle formation is a complex process that involves a series of steps during embryogenesis, including the activation of satellite cells into muscle precursor cells, the withdrawal of myoblasts from the cell cycle, the differentiation of myoblasts into myotubes, and forming the mature muscle fibers
In order to further explore the localization of gga-miR-2954 in chicken muscle tissue, its expression in E13.5 male chicken leg muscle was from embryonic Day 7 (E7) to D1 was measured by Q‐PCR
We conduct a series of experiments, and the results show that gga-miR-2954 can inhibit myoblast proliferation and promote the expression of myogenic genes MyoG and MyHC, which differentiate into myofibers [35], we speculate that in the process of muscle fiber formation in the chicken embryo, gga-miR-2954 might accelerate the differentiation process of myoblasts by allowing myoblasts to exit from the cell cycle and quickly enter into the differentiation process
Summary
Skeletal muscle formation is a complex process that involves a series of steps during embryogenesis, including the activation of satellite cells into muscle precursor cells (myoblasts), the withdrawal of myoblasts from the cell cycle, the differentiation of myoblasts into myotubes, and forming the mature muscle fibers. Besides MRFs and MEF2, many other factors and signal pathways are involved in this regulation process, for example, the PAX and Six family, the signal pathways of Wnt, TGF-β and Notch, and non-coding RNA like lncRNAs (Long noncoding RNAs) and miRNAs (MicroRNAs) [5] These regulatory factors interact with each other, and form a comprehensive network to regulate muscle development in an orderly manner. Recent studies have shown that miR-223 [13] and miR-133a-3p [14] can accelerate chicken primary myoblast from the proliferative phase to the differentiation phase, whereas, miR-203 [15], miR-16-5p [16], miR-133a-5p/miR-29b1-5p [17] and miR-130b-3p/301b-3p [18] inhibits myoblast differentiation These results further revealed the wide participation of miRNAs in the regulation of the growth and development of skeletal muscles
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