Epigenetic Regulation Mechanisms of the Cofilin-1 Gene in the Development and Differentiation of Bovine Primary Myoblasts

Abstract

As the quality of beef products has received increasing attention, it is essential to explore the underlying transcriptional and epigenetic mechanisms of meat traits. Our project uses Qinchuan cattle as the research subject. First, we examined the spatiotemporal expression pattern of the CFL1 gene in a panel of fetal bovine, calf, and adult cattle samples. Then, we performed DNA methylation experiments of CFL1 on myogenesis and muscle maturation using the BSP amplification and COBRA sequencing techniques and found that high DNA methylation levels showed low expression levels. Next, we performed an assay between bta-miR-182 and the CFL1 gene and demonstrated that miR-182 could promote bovine primary myoblast differentiation by negatively regulated the expression of CFL1. Finally, we constructed an adenovirus overexpression and interference vector and found that CFL1 could suppress the differentiation of bovine primary myoblasts. In summary, our experiment comprehensively analyzes the epigenetic regulation mechanisms of the CFL1 gene in the development and differentiation of bovine primary myoblasts. This has far-reaching significance for improving the meat production and meat quality of Qinchuan cattle. This can provide reliable data support and a theoretical research basis for the rapid and efficient breeding selection of local yellow cattle and the genetic improvement of meat quality.

New Insight into Muscle-Type Cofilin (CFL2) as an Essential Mediator in Promoting Myogenic Differentiation in Cattle

“…Recent evidence indicates that cofilin family genes ( CFL1 and CFL2 ) participate in actin cytoskeleton remodeling [ 40 ]. Notably, the CFL1 and CFL2 genes can be expressed simultaneously in embryonic skeletal muscle, whereas the CFL2 gene replaces the CFL1 gene in the late stage of differentiation [ 6 , 8 ]. This means that cofilin only presents one subtype ( CFL1 or CFL2 ) along with the muscle cell growth and development in healthy mouse [ 41 ].…”

“…Primary bovine myoblasts (PBMs) were harvested from fetal bovine longissimus dorsi muscle and cultured by collagenase-I digestion as previously described [ 13 ]. Cell lines and PBMs were cultured following our previously established protocols [ 6 ].…”

“…In healthy mouse muscle cells, the type of cofilin from non-muscle cofilin transits to muscle cofilin [ 5 ]. In our previous study, we found that CFL1-mediated epigenetic regulation mechanisms were involved in the myogenic growth and differentiation [ 6 ]. Muscle-type cofilin is a new member of the cofilin protein family, which mainly regulates the expression of skeleton muscle and cardiac muscle.…”

Global DNA Methylation, miRNA, and mRNA Profiles in Sheep Skeletal Muscle Promoted by Hybridization

“…Other studies have reported DNA methylation in the muscle of fetal and adult Hu sheep, highlighting ADIPOQ (adiponectin, C1Q and collagen domain containing), CCNA2 (cyclin A2), ITGA2 (integrin subunit alpha 2), and MYOG (myogenin) as mediators of muscle growth and metabolism . Nevertheless, meat quality is a very complex trait, determined by multiple regulatory mechanisms, , such as epigenetic marks/factors, , miRNAs, translation regulation, RNA/peptide degradation, and post-translational modification . Therefore, the combined analysis of the transcriptome, small noncoding RNAs, and DNA methylation can provide a comprehensive understanding of the molecular mechanisms that determine meat quality.…”

Identification cloning and functional analysis of novel natural antisense lncRNA CFL1-AS1 in cattle

“…Both the CFL2 gene and CFL1-AS1 parent gene CFL1 are closely related to muscle growth and development and play an important role in normal muscle function and regeneration. Previous studies have shown that the CFL1 and CFL2 genes are specifically expressed in different growth and development stages of bovine muscle and show opposite expression patterns during muscle cell differentiation [ 17 , 18 ]. RNAhybrid analysis identified a miR-125b binding site on CFL1-AS1, and studies have shown that miR-125b can regulate the proliferation and differentiation of myoblasts [ 19 ].…”