草鱼两个肌肉生长抑制素cDNA克隆、表达及过量表达对胚胎发育的影响  

Two cDNAs Cloning,Expression and Overexpression in Embryo of Myostatin from Grass Carp(Ctenopharyngodon idellus)

作　　者：濮剑威[1] 孙成飞[1] 蒋霞云[1] 邹曙明[1] 

机构地区：[1]上海海洋大学农业部种质资源与利用重点开放实验室,上海201306

出　　处：《生物技术通报》2011年第8期153-160,166,共9页Biotechnology Bulletin

基　　金：国家高技术研究发展计划"863"计划项目(2009AA10Z105);公益性行业科研专项(200903045);上海市教委曙光计划项目(08SG50)

摘　　要：肌肉生长抑制素(MSTN)是抑制肌肉生长和发育的重要生长调控因子。通过cDNA末端快速扩增法(RACE)克隆草鱼MSTN-1型和MSTN-2型全长cDNA。RT-PCR分析结果表明,MSTN-1在草鱼肌肉、脑和眼中的转录量较高,在肝胰脏、脾脏和心的转录量较低,在肠、腮、性腺和肾中无表达;MSTN-2只在脑和肌肉中有表达。在草鱼胚胎发育的0-36 hpf期间,MSTN-1的转录量较低;在胚胎发育的36-48 hpf期间,其转录量呈逐渐升高的趋势;MSTN-2各时相均无表达,可能因为该基因在草鱼胚胎发育过程中不起重要作用。通过分别显微注射MSTN-1型和MSTN-2型mRNA至斑马鱼1-2细胞期胚胎。结果显示,注射MSTN-1型mRNA过表达可导致斑马鱼体节发生期胚胎的前-后轴拉长,背-腹轴变短,脊索轻微扭曲,以及体节发育受到强烈抑制而不分化等现象。注射MSTN-2型mRNA胚胎早期发育有所延迟并未发生明显变化,但发育至60 h之后尾部明显发生严重弯曲。Myostatin（MSTN）is an important negative regulator of myogenesis,which can inhibit myoblast proliferation and differentiation.In this study,we identified two myostatin full-length cDNAs in grass crap（Ctenopharyngodon idellus）,by rapid amplification of cDNA ends（RACE）.RT-PCR analysis demonstrates that MSTN are extensively expressed in different tissues of grass carp.MSTN-1 from the muscles,brain and eyes showed high levels of transcription,while the transcript levels in liver pancreas,spleen and heart was low;there was no expression in intestines,gills,gonads and kidney.MSTN-2 was only expressed in brain and muscle.During embryos development,the mRNA expression of MSTN-1 was low from 0 hpf to 36 hpf,whereas the expression level increases gradually from 36 hpf to 48 hpf;and MSTN-2 expression was low.By microinjection,overexpression of MSTN-1,and MSTN-2 mRNA to 1-2 cell stage embryos of grass carp,overexpression of MSTN-1 mRNA in embryos can cause embryo abnormal,which resulted in elongated anterior-posterior axis,shorter dorsal-ventral axis,slightly distorted notochord,and strongly inhibited somites.Overexpression of MSTN-2 type mRNA early embryonic development did not change significantly,but the development to 60 hours after the tail was bent.

关 键 词：草鱼 肌肉生长抑制素 克隆 过表达 

分 类 号：S917.4[农业科学—水产科学] Q78[生物学—分子生物学]