Jiang Junfang, Guo Liangyong, Huang Xin, Zheng Kaizhi, He Sangang, Shan Huili
Institute of Animal Husbandry and Veterinary, Zhejiang Academy of Agricultural Sciences, Hangzhou, China.
Huzhou Agricultural Science and Technology Development Center, Institute of animal Science, Huzhou, China.
Front Genet. 2024 Aug 21;15:1449144. doi: 10.3389/fgene.2024.1449144. eCollection 2024.
N6-Methyladenosine (m6A) RNA modification plays an essential role in many biological processes. To investigate the regulatory role of m6A on the skeletal muscle development in Hu sheep, this study took newborn Hu sheep (b_B Group) and six-month-old Hu sheep (s_B Group) as the objects. MeRIP-seq and RNA-Seq analysis techniques were used to detect differentially methylated genes (DMGs) and differentially expressed genes (DEGs) in the longissimus dorsi muscle of Hu sheep at different months of age. Then, conjoint analysis was further employed to screen for key genes involved in skeletal muscle development that are modified by m6A and expressed by mRNA. According to the results of the MeRIP-seq analysis, there were 285 m6A differentially methylated peaks (DMPs) in total between b_B Group and s_B Group, with 192 significant upregulated peaks and 93 significant downregulated peaks. GO and KEGG analysis revealed that DMGs are mainly enriched in actin-binding, cellular transport, and metabolic pathways. According to the results of the RNA-seq analysis, there were 4,349 DEGs in total between b_B Group and s_B Group, with 2010 upregulated genes and 2,339 downregulated genes. DEGs are found to be mainly enriched in the regulation of actin cytoskeleton tissue, AMPK and FoxO signaling pathways, etc. The conjoint analysis demonstrated that 283 genes were both modified by m6A and expressed by mRNA. Among them, three genes relevant to muscle growth (RGMB, MAPK8IP3, and RSPO3) were selected as candidates for quantitative validation, and the results were in line with the sequencing results. The results mentioned above all suggest that m6A plays a certain role in the skeletal muscle development in Hu sheep.
N6-甲基腺苷（m6A）RNA修饰在许多生物学过程中起着至关重要的作用。为了研究m6A对湖羊骨骼肌发育的调控作用，本研究以新生湖羊（b_B组）和6月龄湖羊（s_B组）为研究对象。采用甲基化RNA免疫沉淀测序（MeRIP-seq）和RNA测序（RNA-Seq）分析技术，检测不同月龄湖羊背最长肌中的差异甲基化基因（DMG）和差异表达基因（DEG）。然后，进一步采用联合分析筛选出参与骨骼肌发育的关键基因，这些基因被m6A修饰并通过mRNA表达。根据MeRIP-seq分析结果，b_B组和s_B组之间共有285个m6A差异甲基化峰（DMP），其中192个显著上调峰和93个显著下调峰。基因本体（GO）和京都基因与基因组百科全书（KEGG）分析表明，DMG主要富集于肌动蛋白结合、细胞运输和代谢途径。根据RNA-Seq分析结果，b_B组和s_B组之间共有4349个DEG，其中2010个上调基因和2339个下调基因。发现DEG主要富集于肌动蛋白细胞骨架组织调控、AMPK和FoxO信号通路等。联合分析表明，有283个基因既被m6A修饰又通过mRNA表达。其中，选择了3个与肌肉生长相关的基因（RGMB、MAPK8IP3和RSPO3）作为定量验证的候选基因，结果与测序结果一致。上述结果均表明m6A在湖羊骨骼肌发育中发挥一定作用。
