基于蛋白组和转录组分析肌卫星细胞到肌管转化中的关键基因

doi:10.1016/j.jia.2023.08.001

摘要/Abstract

摘要：

肌纤维是骨骼肌的主要组成部分，由肌管成熟形成。在早期发育过程中，骨骼肌卫星细胞（SSCs）增殖为成肌细胞，随后成肌细胞经历分化和融合形成肌管。然而，从SSCs到肌管转化过渡机制仍不清晰。因此，本研究采用了RNA-seq和DIA技术对山羊肌卫星细胞、成肌细胞（分化2天）和肌管（分化10天）进行了转录组和蛋白组测序。首先，对两个组学分别进行了差异分析，转录组中共鉴定到5785个差异基因，蛋白组中共鉴定到2946个差异蛋白。蛋白质组分析发现SLMAP和STOM可能与肌管的形成有关。沉默SLMAP后，成肌标记基因MyoD的明显上调（P<0.01）和肌管标记基因MyoG和Myosin7明显下调（P<0.01），但Desmin的表达水平没有变化；沉默STOM后，成肌标记基因MyoD的明显上调（P<0.01）和肌管标记基因MyoG、Myosin7和Desmin均明显下调（P<0.01）。在更严格的差异分析条件下（差异蛋白|log2(FC)|>1.2；差异基因|log2(FC)|>2)）整合两个组学数据发现，在肌卫星细胞和成肌细胞比较组中，18个因子呈正相关，37个因子呈负相关；在成肌细胞和肌管比较组中，31个因子呈正相关，10个因子呈负相关。这些因子的GO分析表明，从肌卫星细胞到成肌细胞转变时，分化和迁移相关的因子SVIL、ENSCHIG00000026624（AQP1）、SERPINE1上调，同时伴随着细胞凋亡。在成肌细胞到肌管转变时，与细胞粘附和信号转导有关的候选因子在肌管中高度表达，CCN2、TGFB1、MYL2和MYL4被确定为成肌细胞和肌管比较组的关键候选因子。综上，本研究从转录组和蛋白组中筛选到了可能影响肌卫星细胞到成肌细胞，再到肌管转变的关键因子，对肌肉早期发育或损伤后再生过程中肌管形成提供新的解析。

Abstract:

Muscle fibers are the main component of skeletal muscle and undergo maturation through the formation of myotubes. During early development, a population of skeletal muscle satellite cells (SSCs) proliferate into myoblasts. The myoblasts then undergo further differentiation and fusion events, leading to the development of myotubes. However, the mechanisms involved in the transition from SSCs to myotube formation remain unclear. In this study, we characterized changes in the proteomic and transcriptomic expression profiles of SSCs, myoblasts (differentiation for 2 d) and myotubes (differentiation for 10 d). Proteomic analysis identified SLMAP and STOM as potentially associated with myotube formation. In addition, some different changes in MyoD, MyoG, Myosin7 and Desmin occurred after silencing SLMAP and STOM, suggesting that they may affect changes in the myogenic marker. GO analysis indicated that the differentiation and migration factors SVIL, ENSCHIG00000026624 (AQP1) and SERPINE1 enhanced the transition from SSCs to myoblasts, accompanied by changes in the apoptotic balance. In the myoblast vs. myotube group, candidates related to cell adhesion and signal transduction were highly expressed in the myotubes. Additionally, CCN2, TGFB1, MYL2 and MYL4 were identified as hub-candidates in this group. These data enhance our existing understanding of myotube formation during early development and repair.

Key words: proteome , transcriptome , skeletal muscle satellite cells , myoblast , myotube Introduction

ZHENG Qi, HU Rong-cui, ZHU Cui-yun, JING Jing, LOU Meng-yu, ZHANG Si-huan, LI Shuang, CAO Hong-guo, ZHANG Xiao-rong, LING Ying-hui. 基于蛋白组和转录组分析肌卫星细胞到肌管转化中的关键基因[J]. Journal of Integrative Agriculture, 2023, 22(10): 3135-3147.

ZHENG Qi, HU Rong-cui, ZHU Cui-yun, JING Jing, LOU Meng-yu, ZHANG Si-huan, LI Shuang, CAO Hong-guo, ZHANG Xiao-rong, LING Ying-hui. Identification of transition factors in myotube formation from proteome and transcriptome analyses[J]. Journal of Integrative Agriculture, 2023, 22(10): 3135-3147.

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