Table_4_Buffalo long non-coding RNA gene11007 promotes myoblasts proliferation.DOCX

Buffalo meat is of good quality because it is lean and tender, and could bring significant cardiovascular benefits. The underlying difference in muscle development and meat quality is a complex and precisely orchestrated process which has been demonstrated to be regulated by long non-coding RNAs (lncRNAs). However, the regulatory role of lncRNAs in the growth and development of buffalo skeletal muscle is still unclear. In this study, the Ribo-Zero RNA-Seq method was used to explore the lncRNA expression profiles of buffalo myoblasts during the proliferation and differentiation phases. A specific set of 9,978 lncRNAs was found. By comparing the expression profiles of lncRNAs, it was found that there were 1,576 differentially expressed lncRNAs (DELs) during buffalo myoblast differentiation. Twelve DELs were chosen and subsequently verified in eight different buffalo tissues during fetal and adult stages by using qPCR. Gene11007 was found to be one of the most down-regulated lncRNAs during buffalo myoblasts differentiation and it was subsequently characterized. EdU, CCK-8, qPCR and western blotting assays showed that gene11007 promoted the proliferation of buffalo myoblasts but it had no effect on cell differentiation. Our research may enrich the genome annotations of buffalo and provide a new molecular target for the in-depth understanding of the regulation of lncRNAs in skeletal muscle.

水牛肉因其瘦嫩而品质优良，能够显著提升心血管健康。肌肉发育与肉质差异的形成是一个复杂且精准调控的过程，已被证实受到长非编码RNA（lncRNAs）的调控。然而，lncRNAs在调节水牛骨骼肌生长与发育中的具体作用尚不明确。本研究采用Ribo-Zero RNA-Seq技术，探究了水牛肌原细胞在增殖与分化阶段lncRNA的表达谱。研究发现，存在一组共9,978个lncRNAs。通过比较lncRNA的表达谱，发现在水牛肌原细胞分化过程中有1,576个差异表达lncRNAs（DELs）。选取了12个DELs，并在八个不同水牛组织（胎儿期和成年期）中通过实时荧光定量PCR（qPCR）进行验证。研究发现，基因11007在水牛肌原细胞分化过程中表达下调，并对其进行了一系列特征描述。EdU、CCK-8、qPCR和蛋白质印迹法（Western blotting）等实验结果表明，基因11007能够促进水牛肌原细胞的增殖，但对细胞分化无影响。本研究可能丰富水牛的基因组注释，并为深入理解lncRNAs在骨骼肌中的调控机制提供新的分子靶点。