Abstract
The proliferation and differentiation of chicken primary myoblasts (CPMs) play an important role in the development of skeletal muscle. In our previous research, RNA-seq analysis showed that microRNA-7 (miR-7) was relatively highly expressed in the proliferation phase of CPMs, but its expression level decreased significantly after CPMS-induced differentiation. Meanwhile, the mechanism by which the miR-7 regulates the proliferation and differentiation of CPMs is still unknown. In this study, we found that the expression levels of miR-7 and the Krüppel-like factor 4 (KLF4) gene were negatively correlated during the embryonic phase, and in vitro induced differentiation. A dual-luciferase assay and a rescue experiment show that there is a target relationship between miR-7 and the KLF4 gene. Meanwhile, the results show that overexpression of miR-7 inhibited the proliferation and differentiation of CPMs, while inhibition of miR-7 had the opposite effects. Furthermore, overexpression of the KLF4 gene was found to significantly promote the proliferation and differentiation of CPMs. Conversely, inhibition of the KLF4 gene was able to significantly decrease the proliferation and differentiation of CPMs. Our results demonstrate, for the first time, that miR-7 inhibits the proliferation and differentiation of myoblasts by targeting the KLF4 gene in chicken primary myoblasts.
Highlights
The embryonic period is a critical period for the growth of broilers, and the number of muscle fibers is mostly determined during this period
The results show that the expression level of miR-7 in the chest and leg muscles increased between E12 and day 1 (Figure 1A)
For the Krüppel-like factor 4 (KLF4) gene, the results show that the expression level of KLF4 in the chest and leg muscles decreased between E12 and day 1 (Figure 1C)
Summary
The embryonic period is a critical period for the growth of broilers, and the number of muscle fibers is mostly determined during this period. MiRNAs degrade or inhibit the expression of target genes by pairing with the 3 untranslated region (3 UTR) of the target mRNAs, thereby regulating the expression of the target genes posttranscriptionally (Pasquinelli and Ruvkun, 2002; Sun et al, 2017). These small non-coding RNAs are critical for the regulation of intracellular translational processes and play a key role in several cellular life activities, including differentiation, proliferation, and signal transduction (Stefani and Slack, 2008; Ebert and Sharp, 2012; Gurtan et al, 2012). The way in which miR7 regulates the proliferation and differentiation of CPMs remains unknown
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