[Functional characterization of recombinant myostatin and its inhibitory role to chicken muscle development].

Wei Yang, Kun Wang, Yan Chen, Yong Zhang, Bo Huang, Da-Hai Zhu
{"title":"[Functional characterization of recombinant myostatin and its inhibitory role to chicken muscle development].","authors":"Wei Yang,&nbsp;Kun Wang,&nbsp;Yan Chen,&nbsp;Yong Zhang,&nbsp;Bo Huang,&nbsp;Da-Hai Zhu","doi":"","DOIUrl":null,"url":null,"abstract":"<p><p>Myostatin is a recently discovered member of transforming growth factor beta (TGFbeta) superfamily and shares similar structure features with other members of TGFbeta superfamily. For a better understanding of molecular mechanism of myostatin function, the production of C-terminal truncated form of recombinant myostatin protein (rMSTN) in E. coli was previously reported. Herein, the functional role of the recombinant myostatin in regulating myogenesis in a chicken embryonic myoblasts (CEMs) system was determined. By using flow cytometric analysis, the myostatin was found to inhibit cell cycle transition from G1 to S phase and result in a cell cycle arrest at G1. In addition, myostatin blocked the multi-nucleus myotube formation and caused a decreased expression of the muscle cell differentiation markers (myogenin and MHC) in CEMs. In this study, a rabbit polyclonal antibody against myostatin was produced and high affinity and specificity of this anti-myostatin antibody to recombinant and endogenous myostatin were assayed by Western blot analysis. Further studies showed that the antibody could also recognize the tissue endogenous myostatin of human, mouse and rat. A specific 40 kD band was detected in chicken muscle, which suggested that chicken myostatin might have different splicing pattern. Immunofluorescence assay indicated that myostatin predominantly existed in the cytosol in C2C12 cells. Taken together, the results show that myostatin inhibits chicken muscle cells proliferation and differentiation and down-regulates expression of two differentiation marker gene in CEMs. Remarkably, production of functional recombinant myostatin protein and its specific antibody provides important reagents for unraveling molecular mechanisms underlying myostatin action during myogenesis.</p>","PeriodicalId":21763,"journal":{"name":"Sheng wu hua xue yu sheng wu wu li xue bao Acta biochimica et biophysica Sinica","volume":"35 11","pages":"1016-22"},"PeriodicalIF":0.0000,"publicationDate":"2003-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Sheng wu hua xue yu sheng wu wu li xue bao Acta biochimica et biophysica Sinica","FirstCategoryId":"1085","ListUrlMain":"","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 0

Abstract

Myostatin is a recently discovered member of transforming growth factor beta (TGFbeta) superfamily and shares similar structure features with other members of TGFbeta superfamily. For a better understanding of molecular mechanism of myostatin function, the production of C-terminal truncated form of recombinant myostatin protein (rMSTN) in E. coli was previously reported. Herein, the functional role of the recombinant myostatin in regulating myogenesis in a chicken embryonic myoblasts (CEMs) system was determined. By using flow cytometric analysis, the myostatin was found to inhibit cell cycle transition from G1 to S phase and result in a cell cycle arrest at G1. In addition, myostatin blocked the multi-nucleus myotube formation and caused a decreased expression of the muscle cell differentiation markers (myogenin and MHC) in CEMs. In this study, a rabbit polyclonal antibody against myostatin was produced and high affinity and specificity of this anti-myostatin antibody to recombinant and endogenous myostatin were assayed by Western blot analysis. Further studies showed that the antibody could also recognize the tissue endogenous myostatin of human, mouse and rat. A specific 40 kD band was detected in chicken muscle, which suggested that chicken myostatin might have different splicing pattern. Immunofluorescence assay indicated that myostatin predominantly existed in the cytosol in C2C12 cells. Taken together, the results show that myostatin inhibits chicken muscle cells proliferation and differentiation and down-regulates expression of two differentiation marker gene in CEMs. Remarkably, production of functional recombinant myostatin protein and its specific antibody provides important reagents for unraveling molecular mechanisms underlying myostatin action during myogenesis.

分享 分享
微信好友 朋友圈 QQ好友 复制链接
本刊更多论文
重组肌生长抑制素的功能表征及其对鸡肌肉发育的抑制作用。
肌生长抑制素是最近发现的转化生长因子β (tgfβ)超家族成员,与tgfβ超家族的其他成员具有相似的结构特征。为了更好地了解肌生长抑制素功能的分子机制,以前有报道在大肠杆菌中产生c端截断形式的重组肌生长抑制素蛋白(rMSTN)。本研究确定了重组肌生长抑制素在鸡胚成肌细胞(CEMs)系统中调控肌肉发生的功能作用。通过流式细胞术分析,发现肌生长抑制素抑制细胞周期从G1期向S期转变,导致细胞周期阻滞在G1期。此外,肌生长抑制素阻断了多核肌管的形成,并导致CEMs中肌肉细胞分化标志物(肌生成素和MHC)的表达降低。本研究制备了兔肌生长抑制素多克隆抗体,并通过Western blot检测了该抗体对重组肌生长抑制素和内源性肌生长抑制素的高亲和力和特异性。进一步研究表明,该抗体还能识别人、小鼠和大鼠的组织内源性肌生成抑制素。在鸡肌肉中检测到一个特异性的40 kD条带,表明鸡肌肉生长抑制素可能具有不同的剪接模式。免疫荧光分析表明C2C12细胞的胞浆中主要存在肌生长抑制素。综上所述,肌生长抑制素抑制了鸡CEMs中肌细胞的增殖和分化,并下调了两种分化标志基因的表达。值得注意的是,功能性重组肌生长抑制素蛋白及其特异性抗体的产生为揭示肌生成过程中肌生长抑制素作用的分子机制提供了重要的试剂。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
求助全文
约1分钟内获得全文 去求助
来源期刊
自引率
0.00%
发文量
0
期刊最新文献
Analysis of gene expression in hepatitis B virus transfected cell line induced by interferon. [Cloning and expression of tumor necrosis factor (TNFalpha) cDNA from red seabream pagrus major]. [Effects of sodium selenite on telomerase activity and telomere length]. [Cloning, eukaryotic expression and function assay of recombinant leukemia inhibitory factor gene LIF]. [The immunopotentiation of human B lymphocyte stimulator C-terminal peptide].
×
引用
GB/T 7714-2015
复制
MLA
复制
APA
复制
导出至
BibTeX EndNote RefMan NoteFirst NoteExpress
×
×
提示
您的信息不完整,为了账户安全,请先补充。
现在去补充
×
提示
您因"违规操作"
具体请查看互助需知
我知道了
×
提示
现在去查看 取消
×
提示
确定
0
微信
客服QQ
Book学术公众号 扫码关注我们
反馈
×
意见反馈
请填写您的意见或建议
请填写您的手机或邮箱
已复制链接
已复制链接
快去分享给好友吧!
我知道了
×
扫码分享
扫码分享
Book学术官方微信
Book学术文献互助
Book学术文献互助群
群 号:481959085
Book学术
文献互助 智能选刊 最新文献 互助须知 联系我们:info@booksci.cn
Book学术提供免费学术资源搜索服务,方便国内外学者检索中英文文献。致力于提供最便捷和优质的服务体验。
Copyright © 2023 Book学术 All rights reserved.
ghs 京公网安备 11010802042870号 京ICP备2023020795号-1