Abstract
Background: Porcine skeletal muscle satellite cells play important roles in myogenesis and muscle regeneration. Integrated analysis of transcriptome and histone modifications would reveal epigenomic roles in promoting myogenic differentiation in swine. Methods: Porcine satellite cells (PSCs) were isolated and in-vitro cultured from newborn piglets. RNA Sequencing (RNA-Seq) and Chromatin Immunoprecipitation Sequencing (ChIP-Seq) experiments were performed using proliferating cells and terminal myotubes in order to interrogate the transcriptomic profiles, as well as the distribution of histone markers—H3K4me3, H3K27me3, and H3K27ac—and RNA polymerase II. Results: The study identified 917 differentially expressed genes during cell differentiation. The landscape of epigenetic marks was displayed on a genome-wide scale, which had globally shrunken. H3K27me3 reinforcement participated in obstructing the transcription of proliferation-related genes, while its depletion was closely related to the up-regulation of myogenic genes. Furthermore, the degree of H3K27me3 modification was dramatically reduced by 50%, and 139 myogenic genes were upregulated to promote cell differentiation. Conclusions: The depletion of H3K27me3 was shown to promote porcine satellite cell differentiation through upregulating the transcription level of myogenic genes. Our findings in this study provide new insights of the epigenomic mechanisms occurring during myogenic differentiation, and shed light on chromatin states and the dynamics underlying myogenesis.
Highlights
Porcine skeletal muscle is an important source of protein, and a model for studying human muscle-related diseases [1,2]
The PRO and D2 cells were characterized by PAX7 immunofluorescent cell (IFC) staining, which indicated the attribution of satellite cells
H3K4me3, H3K27ac, and H3K27me3, together with RNA polymerase II (RNAPII), were interrogated in myoblasts and myotubes to illustrate the relationship between transcriptomic profiles and histone modifications during differentiation
Summary
Porcine skeletal muscle is an important source of protein, and a model for studying human muscle-related diseases [1,2]. Epigenetic modifications like histone modifications are closely related to the transcriptional regulation of myogenic genes [3,11]. In mouse C2C12 cells, H3K27me histone modification regulates myogenic differentiation of satellite cells by regulating the expression of myogenic transcription factors, such as Myog, while H3K4me at the transcription initiation sites of Myf and MyoD would respectively promote the proliferation and differentiation in C2C12 cells [12,13,14]. Porcine skeletal muscle satellite cells play important roles in myogenesis and muscle regeneration. The degree of H3K27me modification was dramatically reduced by 50%, and 139 myogenic genes were upregulated to promote cell differentiation. Conclusions: The depletion of H3K27me was shown to promote porcine satellite cell differentiation through upregulating the transcription level of myogenic genes. Our findings in this study provide new insights of the epigenomic mechanisms occurring during myogenic differentiation, and shed light on chromatin states and the dynamics underlying myogenesis
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