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| Use of transcriptome sequencing to explore the effect of CSRP3 on chicken myoblasts |
| SHAN Yan-ju1, JI Gai-ge1, ZHANG Ming1, LIU Yi-fan1, 2, TU Yun-jie1, JU Xiao-jun1, SHU Jing-ting1#, ZOU Jian-min1 |
1 Key Laboratory for Poultry Genetics and Breeding of Jiangsu Province, Poultry Institute, Chinese Academy of Agricultural Sciences, Yangzhou 225125, P.R.China
2 Joint International Research Laboratory of Agriculture and Agri-Product Safety of Ministry of Education, Yangzhou University, Yangzhou 225009, P.R.China
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摘要
调节鸡肌纤维类型特异性和维持性的机制还很不清楚。在哺乳动物中，CSRP3在维持典型的肌肉结构和功能方面起着非常重要的作用。本研究探讨了CSRP3在鸡骨骼肌中的作用。首先，制备了鸡CSRP3抗体，比较了四种不同肌纤维组成的鸡骨骼肌中CSRP3基因的mRNA和蛋白表达水平。然后分析CSRP3沉默后对鸡成肌细胞转录组表达谱的影响。结果表明，鸡骨骼肌中CSRP3基因的mRNA和蛋白表达水平均与其肌纤维类型组成相关。以Q<0.05，差异倍数大于1.5为筛选条件，共鉴定出650个差异基因。CSRP3沉默后，255个基因表达上调，395个基因表达下调。功能富集分析结果表明，心肌细胞肾上腺素能信号、脂肪细胞因子信号和apelin信号等多种通路在差异表达基因和所有表达基因中均显著（P<0.05）富集。共表达基因网络结果表明，CSRP3沉默后引起与肌原纤维组装、肌肉分化和收缩相关基因的代偿性上调（Q<0.05）。同时，CSRP3沉默后，两个快速肌球蛋白重链基因（MyH1B和MyH1E）表达上调（Q<0.05）。这些结果表明，CSRP3在鸡肌纤维组成中发挥着重要作用，并可能通过调节MyH1B和MyH1E的表达从而影响鸡肌纤维类型的分布。
Abstract
The mechanisms that regulate the specificity and maintenance of chicken muscle fiber types remain largely unknown. In mammals, CSRP3 has been shown to play a vital role in the maintenance of typical muscle structure and function. This study investigated the role that CSRP3 plays in chicken skeletal muscle. First, the antibody against chicken CSRP3 protein was prepared, and the expression levels of the mRNA and protein of the CSRP3 gene in four chicken skeletal muscles with different myofiber compositions were compared. Then the effects of CSRP3 silencing on the expression profile of chicken myoblast transcriptomes were analyzed. The results showed that the expression levels of the mRNA and protein of the CSRP3 gene were both associated with the composition of fiber types in chicken skeletal muscles. A total of 650 genes with at least 1.5-fold differences (Q<0.05) were identified, of which 255 genes were upregulated and 395 genes were downregulated by CSRP3 silencing. Functional enrichment showed that several pathways, including adrenergic signaling in cardiomyocytes, adipocytokine signaling pathway and apelin signaling pathway, were significantly (P<0.05) enriched both in differentially expressed genes and all expressed genes. The co-expressed gene network suggested that CSRP3 silencing caused a compensatory upregulation (Q<0.05) of genes related to the assembly of myofibrils, muscle differentiation, and contraction. Meanwhile, two fast myosin heavy chain genes (MyH1B and MyH1E) were upregulated (Q<0.05) upon CSRP3 silencing. These results suggested that CSRP3 plays a crucial role in chicken myofiber composition, and affects the distribution of chicken myofiber types, probably by regulating the expression of MyH1B and MyH1E.
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Received: 15 October 2021
Accepted: 05 May 2022
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Fund: This work was supported by the earmarked fund for China Agriculture Research System (CARS-41), the earmarked fund for Jiangsu Agricultural Industry Technology System, China (JATS[2021]396), the Special Fund for Major Breeding Programs in Jiangsu Province (PZCZ201728), the Natural Science Foundation of Jiangsu Province (BK20161322, BK20211121, and BK20210955), the Projects of Key Laboratory for Poultry Genetics and Breeding of Jiangsu Province (JQLAB-ZZ-201703), the Open Project Program of Joint International Research
Laboratory of Agriculture and Agri-Product Safety of the Ministry of Education, Yangzhou University, China (JILAR-KF202020), the Yangzhou Science and Technology Support Program for Modem Agriculture (YZ2021029), the Jiangsu Provincal Agricultural Independent Innovation Fund Project (CX(21)2011-1),
and the Independent Scientific Foundation of Public Welfare Scientific Institutes of Jiangsu Province (BM2018026).
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| About author: SHAN Yan-ju, Tel: +86-514-85599076, E-mail: shanyanjusyj @163.com; #Correspondence SHU Jing-ting, Tel: +86-514-85599076, E-mail: shujingting@163.com |
Cite this article:
SHAN Yan-ju, JI Gai-ge, ZHANG Ming, LIU Yi-fan, TU Yun-jie, JU Xiao-jun, SHU Jing-ting, ZOU Jian-min.
2023.
Use of transcriptome sequencing to explore the effect of CSRP3 on chicken myoblasts. Journal of Integrative Agriculture, 22(4): 1159-1171.
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