Institute of Animal Sciences, Chinese Academy of Agricultural Sciences, Key Laboratory of Animal (Poultry) Genetics Breeding and Reproduction, Ministry of Agriculture and Rural Affairs, Beijing, China.
Cell Prolif. 2022 May;55(5):e13219. doi: 10.1111/cpr.13219. Epub 2022 Apr 1.
Although major advances have been made in bovine epigenome study, the epigenetic basis for fetal skeletal muscle development still remains poorly understood. The aim is to recapitulated the time course of fetal skeletal muscle development in vitro, and explore the dynamic changes of chromatin accessibility and gene expression during bovine myoblasts proliferation and differentiation.
PDGFR- cells were isolated from bovine fetal skeletal muscle, then cultured and induced myogenic differentiation in vitro in a time-course study (P, D0, D2,and D4). The assay for transposase-accessible chromatin sequencing (ATAC-seq) and RNA sequencing (RNA-seq) were performed.
Among the enriched transcriptional factors with high variability, we determined the effects of MAFF, ZNF384, and KLF6 in myogenesis using RNA interference (RNAi). In addition, we identified both stage-specific genes and chromatin accessibility regions to reveal the sequential order of gene expression, transcriptional regulatory, and signal pathways involved in bovine skeletal muscle development. Further investigation integrating chromatin accessibility and transcriptome data was conducted to explore cis-regulatory regions in line with gene expression. Moreover, we combined bovine GWAS results of growth traits with regulatory regions defined by chromatin accessibility, providing a suggestive means for a more precise annotation of genetic variants of bovine growth traits.
Overall, these findings provide valuable information for understanding the stepwise regulatory mechanisms in skeletal muscle development and conducting beef cattle genetic improvement programs.
尽管在牛的表观基因组研究方面取得了重大进展，但胎儿骨骼肌发育的表观遗传基础仍知之甚少。本研究旨在体外再现胎儿骨骼肌发育的时间过程，并探讨在牛肌细胞增殖和分化过程中染色质可及性和基因表达的动态变化。
从牛胎儿骨骼肌中分离 PDGFR-细胞，然后进行体外培养和诱导肌源性分化的时间过程研究（P、D0、D2 和 D4）。进行转座酶可及染色质测序（ATAC-seq）和 RNA 测序（RNA-seq）分析。
在高变异性的丰富转录因子中，我们使用 RNA 干扰（RNAi）确定了 MAFF、ZNF384 和 KLF6 在肌生成中的作用。此外，我们还鉴定了阶段特异性基因和染色质可及性区域，以揭示参与牛骨骼肌发育的基因表达、转录调控和信号通路的顺序。进一步整合染色质可及性和转录组数据的研究，以探索与基因表达一致的顺式调控区域。此外，我们将牛生长性状的 GWAS 结果与染色质可及性定义的调控区域相结合，为更精确地注释牛生长性状的遗传变异提供了一种有价值的方法。
总的来说，这些发现为理解骨骼肌发育的逐步调控机制以及开展肉牛遗传改良计划提供了有价值的信息。
