团头鲂lpin1基因的克隆、表达及对肌肉损伤的响应
Cloning and expression of lpin1 gene and its response to muscle injury in Megalobrama amblycephala
金叶子 1石林林 1刘红 1王焕岭1
作者信息
- 1. 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
- 折叠
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 amblycepha-la)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 Mega-lobrama 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.am-blycephala 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.Phy-logenetic 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.am-blycephala.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,respective-ly.The above results indicated that lpin1 gene may play an important role in muscle injury and repair of M.amblycephala.
关键词
团头鲂/lpin1基因/基因表达/肌肉损伤修复Key words
Megalobrama amblycephala/lpin1/gene expression/muscle damage and repair引用本文复制引用
出版年
2024
NETL
NSTL
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