团头鲂lpin1基因的克隆和表达及其对肌肉损伤的响应研究
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华中农业大学水产学院

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国家重点基础研究发展计划(973计划)


Cloning and expression of lpin1 gene and its response to muscle injury in Megalobrama amblycephala
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College of Fishery, Huazhong Agricultural University

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    摘要:

    为研究团头鲂(Megalobrama amblycephala)脂素1(Lipin1,lpin1)基因的序列特征、表达模式及肌肉损伤对其表达的影响,本研究采用荧光定量PCR和整胚原位杂交技术检测了团头鲂lpin1基因的时空表达及肌肉损伤后该基因的表达变化。结果显示团头鲂lpin1基因的ORF序列长2694 bp,编码897个氨基酸,编码的蛋白质具有高度保守的Lipin-N、Lipin-mid和LNS2(Lipin/Ned1/Smp2)结构域,系统进化树分析显示团头鲂Lpin1与鲤和斑马鱼的亲缘关系最近。荧光定量PCR结果表明,lpin1在团头鲂胚胎发育过程中的各个时期均有表达,且在成鱼肌肉组织中表达量最高;整胚原位杂交进一步显示lpin1在肌肉效应期和心跳期的头部和体节中特异表达;对团头鲂幼鱼肌肉进行不同程度损伤后,与未损伤组相比,lpin1基因分别在轻度损伤后48 h和重度损伤后72 h的表达量最高,pax7基因表达量分别在轻度损伤后144 h和重度损伤后96 h最高,由此我们推断lpin1基因在团头鲂肌肉损伤和修复的过程中扮演着重要的角色。

    Abstract:

    To study the sequence and expression characteristics of Lipin1 (lpin1) gene of Megalobrama amblycephala and the effect of muscle injury on its expression, in this study, the temporal and spatial expression of lpin1 gene and its expression change after muscle injury were detected by fluorescent quantitative PCR and whole embryo in situ hybridization. The results showed that the ORF sequence of lpin1 gene was 2694 bp, encoding 897 amino acids. Functional domain analysis showed that this protein had highly conserved Lipin-N, Lipin-mid and LNS2 (Lipin/Ned1/Smp2) domains. Phylogenetic tree analysis showed that Lpin1 was closely related to Cyprinus carpio and Danio rerio. The results of quantitative PCR showed that lpin1 was expressed at all stages during the embryonic development, and the expression level was the highest in the muscle of adult M. amblycephala. Whole embryo in situ hybridization further revealed that lpin1 was specifically expressed in the head and body segments during muscle functioning stage and heartbeating stage. Compared with the uninjured group, the lpin1 expression was significantly increased at 48 h after mild injury and 216 h after severe injury, the pax7 expression was significantly increased at 144 h after mild injury and 96 h after severe injury. We conclude that lpin1 gene may play an important role in muscle injury and repair of M.amblycephala.

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  • 收稿日期:2023-03-16
  • 最后修改日期:2023-08-28
  • 录用日期:2024-08-11
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