Abstract
The trimethylation of histone H3 lysine 27 (H3K27me3) is one of the most important chromatin modifications, which is generally presented as a repressive mark in various biological processes. However, the dynamic and global-scale distribution of H3K27me3 during porcine embryonic muscle development remains unclear. Here, our study provided a comprehensive genome-wide view of H3K27me3 and analyzed the matching transcriptome in the skeletal muscles on days 33, 65, and 90 post-coitus from Duroc fetuses. Transcriptome analysis identified 4,124 differentially expressed genes (DEGs) and revealed the key transcriptional properties in three stages. We found that the global H3K27me3 levels continually increased during embryonic development, and the H3K27me3 level was negatively correlated with gene expression. The loss of H3K27me3 in the promoter was associated with the transcriptional activation of 856 DEGs in various processes, including skeletal muscle development, calcium signaling, and multiple metabolic pathways. We also identified for the first time that H3K27me3 could enrich in the promoter of genes, such as DES, MYL1, TNNC1, and KLF5, to negatively regulate gene expression in porcine satellite cells (PSCs). The loss of H3K27me3 could promote muscle cell differentiation. Taken together, this study provided the first genome-wide landscape of H3K27me3 in porcine embryonic muscle development. It revealed the complex and broad function of H3K27me3 in the regulation of embryonic muscle development from skeletal muscle morphogenesis to myofiber maturation.
Highlights
Skeletal muscles represent the most abundant tissue of the body, and such muscles are essential for motion and support (Chal and Pourquié, 2017)
We further hierarchically clustered the expression across three stages and found that all differentially expressed genes (DEGs) were primarily clustered into two groups: (1) high expression in d33 and (2) commonly high expression in d65 and d90 (Figure 1D)
The expression of DEGs slightly changed from d65 to d90, whereas 23 genes drastically changed between 33 days and 65 days (Supplementary Table S3)
Summary
Skeletal muscles represent the most abundant tissue of the body, and such muscles are essential for motion and support (Chal and Pourquié, 2017). The continual advances of high-throughput sequencing methods in epigenetics allow lower input material and more accurate examination of the dynamic alterations to histone, DNA, and RNA methylation patterns (Barski et al, 2007; Dominissini et al, 2012; Wang et al, 2013; Abo Alrob and Lopaschuk, 2014) These techniques with increasingly analytical tools have contributed to the investigation of RNA splicing and transcriptional regulation in various processes, and the prediction and therapy of diseases (Heerboth et al, 2014; Gu et al, 2015; Bauer et al, 2016). We performed RNA-seq and chromatin immunoprecipitation (ChIP)-seq to provide the first comprehensive analysis of H3K27me and gene expression profile during embryonic muscle development (d33, d65, and d90) in Duroc pig. This study can provide the basis for studying and expanding the regulatory mechanisms of muscle fiber development
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