团头鲂lpin1基因的克隆、表达及对肌肉损伤的响应
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作者单位:

华中农业大学水产学院/农业农村部淡水生物繁育重点实验室,武汉 430070

作者简介:

金叶子,E-mail: 360132041@qq.com

通讯作者:

王焕岭,E-mail: huanlingok@mail.hzau.edu.cn

中图分类号:

Q959.46+8

基金项目:

国家重点研发计划项目(2022YFF1000301);中央高校科研业务费专项(2662022SCYJ005)


Cloning and expression of lpin1 gene and its response to muscle injury in Megalobrama amblycephala
Author:
Affiliation:

College of Fisheries, Huazhong Agricultural University/Key Laboratory of Freshwater Biology and Breeding, Ministry of Agriculture and Rural Affairs, Wuhan 430070,China

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    摘要:

    为深入研究lpin1基因在鱼类肌肉损伤修复过程中的作用机制,以团头鲂(Megalobrama amblycephala)为研究对象,采用荧光定量PCR和整胚原位杂交技术检测团头鲂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2 694 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结果显示,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。以上结果表明,lpin1基因可能在团头鲂肌肉损伤和修复的过程中扮演着重要角色。

    Abstract:

    To study the role of lipin1 (lpin1) gene in the repair of muscle injury in fish, taking Megalobrama amblycephala as the research object, in this study, the temporal and spatial expression of lpin1 gene and its expression change after muscle injury were detected by fluorescent quantitative PCR (qPCR) and whole embryo in situ hybridization (WISH). The results showed that the ORF sequence of the M. amblycephala lpin1 gene was 2 694 bp, encoding 897 amino acids. Functional domain analysis showed that the Lpin1 protein had highly conserved Lipin-N, Lipin-mid and LNS2 (Lipin/Ned1/Smp2) domains. Phylogenetic analysis revealed that the M. amblycephala Lpin1 was closely related to those of Cyprinus carpio and Danio rerio. The results of qPCR showed that lpin1 was expressed at all stages during the embryonic development of M. amblycephala, and the expression level was the highest in the muscle of adult M. amblycephala. The results of WISH 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 the highest at 48 h after mild injury and 216 h after severe injury, respectively, while the pax7 expression was the highest at 144 h after mild injury and 96 h after severe injury, respectively. The above results indicated that lpin1 gene may play an important role in muscle injury and repair of M. amblycephala.

    图1 Lpin1的系统进化分析Fig.1 Phylogenetic analysis of Lpin1
    图2 Lpin1蛋白的氨基酸序列多重比对Fig.2 Multiple alignment and analysis of amino acid sequences of Lpin1 proteins
    图3 lpin1基因在团头鲂成鱼各组织中的相对表达量Fig.3 The relative expression of lpin1 in adult M. amblycephala tissues
    图4 lpin1基因在团头鲂胚胎各发育阶段的相对表达量Fig.4 Relative expression of lpin1 at M. amblycephala different embryo development stages
    图5 原位杂交技术检测团头鲂lpin1基因在胚胎不同发育时期的表达Fig.5 Expression of lpin1 gene at different embryo development stages of M. amblycephala based on in situ hybridization
    图6 团头鲂lpin1(A)和pax7(B)基因在肌肉损伤修复过程中的表达变化Fig.6 Expression changes of lpin1 (A)and pax7(B) genes during muscle repair of M. amblycephala
    表 1 所用引物序列Table 1 Primer sequences used in this study
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金叶子,石林林,刘红,王焕岭.团头鲂lpin1基因的克隆、表达及对肌肉损伤的响应[J].华中农业大学学报,2024,43(5):205-212

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  • 收稿日期:2023-08-28
  • 在线发布日期: 2024-10-08
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