Key Laboratory of Bio-Resources and Eco-Environment of Ministry of Education, College of Life Sciences, Sichuan University, Chengdu 610065, PR China.
National Engineering Laboratory for Animal Breeding and MOA Key Laboratory of Animal Genetics and Breeding, College of Animal Science and Technology, China Agricultural University, Beijing, PR China.
Poult Sci. 2024 Oct;103(10):104097. doi: 10.1016/j.psj.2024.104097. Epub 2024 Jul 14.
Myocyte enhancer factor 2A (MEF2A) is a transcription factor that plays a critical role in cell proliferation, differentiation and apoptosis. In contrast to the wide characterization of its regulation mechanism in mammalian skeletal muscle, its role in chickens is limited. Especially, its wide target genes remain to be identified. Therefore, we utilized Cleavage Under Targets and Tagmentation (CUT&Tag) technology to reveal the genome-wide binding profile of MEF2A in chicken primary myoblasts thus gaining insights into its potential role in muscle development. Our results revealed that MEF2A binding sites were primarily distributed in intergenic and intronic regions. Within the promoter region, although only 8.87% of MEF2A binding sites were found, these binding sites were concentrated around the transcription start site (TSS). Following peak annotation, a total of 1903 genes were identified as potential targets of MEF2A. Gene Ontology (GO) enrichment analysis further revealed that MEF2A target genes may be involved in the regulation of embryonic development in multiple organ systems, including muscle development, gland development, and visual system development. Moreover, a comparison of the MEF2A target genes identified in chicken primary myoblasts with those in mouse C2C12 cells revealed 388 target genes are conserved across species, 1515 target genes are chicken specific. Among these conserved genes, ankyrin repeat and SOCS box containing 5 (ASB5), transmembrane protein 182 (TMEM182), myomesin 2 (MYOM2), leucyl and cystinyl aminopeptidase (LNPEP), actinin alpha 2 (ACTN2), sorbin and SH3 domain containing 1 (SORBS1), ankyrin 3 (ANK3), sarcoglycan delta (SGCD), and ORAI calcium release-activated calcium modulator 1 (ORAI1) exhibited consistent expression patterns with MEF2A during embryonic muscle development. Finally, TMEM182, as an important negative regulator of muscle development, has been validated to be regulated by MEF2A by dual-luciferase and quantitative real-time PCR (qPCR) assays. In summary, our study for the first time provides a wide landscape of MEF2A target genes in chicken primary myoblasts, which supports the active role of MEF2A in chicken muscle development.
肌细胞增强因子 2A（MEF2A）是一种转录因子，在细胞增殖、分化和凋亡中发挥着关键作用。与哺乳动物骨骼肌中其调控机制的广泛特征相比，其在鸡中的作用有限。特别是，其广泛的靶基因仍有待鉴定。因此，我们利用靶向切割和标签技术（CUT&Tag）揭示了 MEF2A 在鸡原代成肌细胞中的全基因组结合谱，从而深入了解其在肌肉发育中的潜在作用。我们的结果表明，MEF2A 结合位点主要分布在基因间区和内含子区。在启动子区域，虽然只发现了 8.87%的 MEF2A 结合位点，但这些结合位点集中在转录起始位点（TSS）周围。峰注释后，共鉴定出 1903 个潜在的 MEF2A 靶基因。基因本体论（GO）富集分析进一步表明，MEF2A 靶基因可能参与多个器官系统的胚胎发育调控，包括肌肉发育、腺体发育和视觉系统发育。此外，将鸡原代成肌细胞中鉴定的 MEF2A 靶基因与小鼠 C2C12 细胞中的靶基因进行比较，发现 388 个靶基因在物种间保守，1515 个靶基因是鸡特有的。在这些保守基因中，ankyrin 重复和 SOCS 盒包含 5（ASB5）、跨膜蛋白 182（TMEM182）、肌联蛋白 2（MYOM2）、亮氨酸和胱氨酸氨肽酶（LNPEP）、肌动蛋白α 2（ACTN2）、sorbin 和 SH3 域包含 1（SORBS1）、ankyrin 3（ANK3）、肌聚糖 delta（SGCD）和钙释放激活钙调节剂 1（ORAI1）在胚胎肌肉发育过程中与 MEF2A 表现出一致的表达模式。最后，跨膜蛋白 182（TMEM182）作为肌肉发育的重要负调控因子，已通过双荧光素酶和实时定量 PCR（qPCR）实验验证其受 MEF2A 调控。综上所述，本研究首次提供了鸡原代成肌细胞中 MEF2A 靶基因的广泛图谱，支持 MEF2A 在鸡肌肉发育中的活跃作用。
