Skip to main content

Advertisement

SpringerLink
Log in

Molecular Cloning and Expression Analysis of the Myostatin Gene in Sea Perch (Lateolabrax japonicus)

  • Original Article
  • Published:
Marine Biotechnology Aims and scope Submit manuscript

Abstract

Myostatin (MSTN) is a member of the transforming growth factor-β (TGF-β) superfamily that functions as a negative regulator of skeletal muscle development and growth in mammals. However, few reports are available about the structure and function of MSTN in teleost. Here, the MSTN gene was cloned from sea perch (Lateolabrax japonicus) by homology cloning and genomic walking. In the 4873-bp genomic sequence, three exons, two introns, and 5′ and 3′ flanking sequences were identified. The sea perch MSTN gene encodes a 374-amino acid protein, including a signal peptide, conserved cysteine residues, and a RXXR proteolytic cleavage domain. Expression analysis of MSTN revealed that MSTN was highly expressed in eyes, brain, and muscle; intermediately in intestine; and weakly in gill, spleen, liver, and heart. It was demonstrated that MSTN mRNA was highly expressed in embryonic stem cell line (LJES1), but it was undetectable in several types of somatic cell lines from sea perch, including fibroblast-like cell, epithelioid cell, and lymphocyte-like cell. Further, it was demonstrated that the 5′ flanking region of the MSTN gene can drive the expression of green fluorescent protein (GFP) reporter gene in LJES1 cells and transgenic zebrafish (Danio rerio). This is the first report on the expression profile of MSTN gene in various types of cell cultures.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6

Similar content being viewed by others

References

  • J Acosta Y Carpio L Borroto O Gonzalez MP Estrada (2005) ArticleTitleMyostatin gene silenced by RNAi show a zebrafish giant phenotype J Biotechnol 119 324–331 Occurrence Handle10.1016/j.jbiotec.2005.04.023

    Article  Google Scholar 

  • SF Altschul W Gish W Miller EW Myers EW Lipman (1990) ArticleTitleBasic local aligment search tool J Mol Biol 215 403–410

    Google Scholar 

  • SF Altschul TL Madden AA Schaffer J Zhang Z Zhang W Miller DJ Lipman (1997) ArticleTitleGapped BLAST and PSI-BLAST: a new generation of protein database search programs Nucl Acids Res 25 3389–3402 Occurrence Handle10.1093/nar/25.17.3389

    Article  Google Scholar 

  • AA Amali CJF Lin YH Chen WL Wang HY Gong CY Lee YL Ko JK Lu GM Her TT Chen JL Wu (2004) ArticleTitleUp-regulation of muscle-specific transcription factors during embryonic somitogenesis of zebrafish (Danio rerio) by knock-down of myostatin-1 Dev Dynam 229 847–856 Occurrence Handle10.1002/dvdy.10454

    Article  Google Scholar 

  • J Bass J Oldham M Sharma R Kambadur (1999) ArticleTitleGrowth factors controlling muscle development Domest Anim Endocrinol 17 191–197 Occurrence Handle10.1016/S0739-7240(99)00036-3

    Article  Google Scholar 

  • PR Biga SB Roberts DB Lliev LAR McCauley JS Moon P Collodi FW Goetz (2005) ArticleTitleThe isolation, characterization, and expression of a novel GDF11 gene and a second myostatin form in zebrafish, Danio rerio Comp Biochem Physiol B 141 218–230 Occurrence Handle10.1016/j.cbpc.2005.03.004

    Article  Google Scholar 

  • M Buckingham (2001) ArticleTitleSkeletal muscle formation in vertebrates Curr Opin Genet Dev 11 440–448 Occurrence Handle10.1016/S0959-437X(00)00215-X

    Article  Google Scholar 

  • SL Chen Y Hong M Schartl (2001) ArticleTitleLack of ultraviolet-light inducibility of the medakafish (Oryzias latipes) tumor suppressor gene p53 Gene 264 197–203 Occurrence Handle10.1016/S0378-1119(01)00340-7

    Article  Google Scholar 

  • SL Chen Y Hong M Schartl (2002) ArticleTitleDevelopment of a positive-negative selection procedure for gene targeting in fish cells Aquaculture 214 67–79 Occurrence Handle10.1016/S0044-8486(01)00811-0

    Article  Google Scholar 

  • SL Chen ZX Sha HQ Ye (2003) ArticleTitleEstablishment of a pluripotent embryonic cell line from sea perch (Lateolabrax japonicus) blastula embryo Aquaculture 218 141–151 Occurrence Handle10.1016/S0044-8486(02)00570-7

    Article  Google Scholar 

  • Chen SL, Sha ZX, Ye HQ, Liu Y, Tian YS, Hong Y, Tang QS (2006) Pluripotency and chimera competence of an embryonic stem cell line from the sea perch (Lateolabrax japonicus). Mar Biotechnol. DOI: 10.1007/s10126-006-6050-1

  • DJ Gregory GC Waldbieser BG Bosworth (2004) ArticleTitleCloning and characterization of myogenic regulatory genes in three Ictalurid species Anim Genet 35 425–430 Occurrence Handle10.1111/j.1365-2052.2004.01193.x

    Article  Google Scholar 

  • L Grobet LJ Martin D Poncelet D Pirottin B Brouwers J Riquet A Schoeberlein S Dunner F Menissier J Massabanda R Fries R Hanset M Georges (1997) ArticleTitleA deletion in the bovine myostatin gene causes the double-muscled phenotype in cattle Nat Genet 17 71–74 Occurrence Handle10.1038/ng0997-71

    Article  Google Scholar 

  • L Grobet D Pirottin F Farnir D Poncelet LJ Royo B Brouwers E Christians D Desmecht F Coignoul R Kahn M Georges (2003) ArticleTitleModulating skeletal muscle mass by postnatal, muscle-specific inactivation of the myostatin gene Genesis 35 227–238 Occurrence Handle10.1002/gene.10188

    Article  Google Scholar 

  • Z Gu Y Zhang P Shi YP Zhang D Zhu H Li (2004) ArticleTitleComparison of avian myostatin genes Anim Genet 35 470–472 Occurrence Handle10.1111/j.1365-2052.2004.01194.x

    Article  Google Scholar 

  • S Ji RL Losinski SG Cornelius GR Frank GM Willis DE Gerrard FF Depreux ME Spurlock (1998) ArticleTitleMyostatin expression in porcine tissues: tissue specificity and developmental and postnatal regulation Am J Physiol 275 1265–1273

    Google Scholar 

  • KA Johansen K Overturf (2005) ArticleTitleQuantitative expression analysis of genes affecting muscle growth during development of rainbow trout (Oncorhynchus mykiss) Mar Biotechnol 7 576–587 Occurrence Handle10.1007/s10126-004-5133-3

    Article  Google Scholar 

  • R Kambadur M Sharma TP Smith JJ Bass (1997) ArticleTitleMutations in myostatin (GDF8) in double-muscled Belgian Blue and Piedmontese cattle Genome Res 7 910–916

    Google Scholar 

  • Ko CF, Chiou TT, Chen TT, Wu JL, Chen JC, Lu JK (2006) Molecular cloning of myostatin gene and characterization of tissue-specific and developmental stage-specific expression of the gene in orange spotted grouper, Epinephelus coioides. Mar Biotechnol 2006 Oct 19 [Epub ahead of print]

  • AM Kocabas H Kucuktas RA Dunham Z Liu (2002) ArticleTitleMolecular characterization and differential expression of the myostatin gene in channel catfish (Ictalurus punctatus) Biochim Biophys Acta 1575 99–107

    Google Scholar 

  • SK Kumar IB Tamura MN Jakobsen (2001) ArticleTitleMEGA2: molecular evolutionary genetics analysis software Bioinformatics 17 1244–1245 Occurrence Handle10.1093/bioinformatics/17.12.1244

    Article  Google Scholar 

  • SH Li YZ Xiong R Zheng AY Li CY Deng SW Jiang MG Lei YQ Wen GC Cao (2002) ArticleTitlePolymorphism of porcine myostatin gene Yi Chuan Xue Bao 29 326–331

    Google Scholar 

  • Liu Y, Sha ZX, Chen SL, Sui SF, Xu MY (2004) Expression of green fluorescent protein gene transferred into newly fertilized sea perch (Lateolabrax japonicus) embryos by microinjection. J Fisf Sci China 11, 385–390 (in Chinese with English abstract)

    Google Scholar 

  • L Maccatrozzo L Bargelloni G Radaelli F Mascarello T Patarnello (2001) ArticleTitleCharacterization of the myostatin gene in the gilthead seabream (Sparus aurata): sequence, genomic structure, and expression pattern Mar Biotechnol 3 224–230 Occurrence Handle10.1007/s101260000064

    Article  Google Scholar 

  • L Maccatrozzo L Bargelloni B Cardazzo G Rizzo T Patarnello (2001) ArticleTitleA novel second myostatin gene is present in teleost fish FEBS Lett 509 36–40 Occurrence Handle10.1016/S0014-5793(01)03124-6

    Article  Google Scholar 

  • L Maccatrozzo L Bargelloni P Patarnello G Radaelli F Mascarello T Patarnello (2002) ArticleTitleCharacterization of the myostatin gene and a linked microsatellite marker in shi drum (Umbrina cirrosa, Sciaenidae) Aquaculture 205 49–60 Occurrence Handle10.1016/S0044-8486(01)00659-7

    Article  Google Scholar 

  • AC McPherron SJ Lee (1997) ArticleTitleDouble muscling in cattle due to mutations in the myostatin gene Proc Natl Acad Sci USA 94 12457–12461 Occurrence Handle10.1073/pnas.94.23.12457

    Article  Google Scholar 

  • AC McPherron AM Lawler SJ Lee (1997) ArticleTitleRegulation of skeletal muscle mass in mice by a new TGF-beta superfamily member Nature 387 83–90 Occurrence Handle10.1038/387083a0

    Article  Google Scholar 

  • TK Ostbye TF Galloway C Nielsen I Gabestad T Bardal O Andersen (2001) ArticleTitleThe two myostatin genes of Atlantic salmon (Salmo salar) are expressed in a variety of tissues Eur J Biochem 268 IssueID20 5249–5257 Occurrence Handle10.1046/j.0014-2956.2001.02456.x

    Article  Google Scholar 

  • PY Rescan (2001) ArticleTitleRegulation and functions of myogenic regulatory factors in lower vertebrates Comp Biochem Physiol B Biochem Mol Biol 130 1–12 Occurrence Handle10.1016/S1096-4959(01)00412-2

    Article  Google Scholar 

  • PY Rescan I Jutel C Ralliere (2001) ArticleTitleTwo myostatin genes are differentially expressed in myotomal muscles of the trout (Oncorhynchus mykiss) J Exp Biol 204 3523–3529

    Google Scholar 

  • SB Roberts FW Goetz (2001) ArticleTitleDifferential skeletal muscle expression of myostatin across teleost species, and the isolation of multiple myostatin isoforms FEBS Lett 491 IssueID3 212–216 Occurrence Handle10.1016/S0014-5793(01)02196-2

    Article  Google Scholar 

  • SB Roberts FW Goetz (2003) ArticleTitleMyostatin protein and RNA transcript levels in adult and developing brook trout Mol Cell Endocrinol 210 9–20 Occurrence Handle10.1016/j.mce.2003.09.002

    Article  Google Scholar 

  • BD Rodgers GM Weber (2001) ArticleTitleSequence conservation among fish myostatin orthologues and the characterization of two additional cDNA clones from Morone saxatilis and Morone americana Comp Biochem Physiol B Biochem Mol Biol 129 597–603 Occurrence Handle10.1016/S1096-4959(01)00350-5

    Article  Google Scholar 

  • BD Rodgers GM Weber CV Sullivan MA Levine (2001) ArticleTitleIsolation and characterization of myostatin complementary deoxyribonucleic acid clones from two commercially important fish: Oreochromis mossambicus and Morone chrysops Endocrinology 142 1412–1418 Occurrence Handle10.1210/en.142.4.1412

    Article  Google Scholar 

  • N Saitou M Nei (1987) ArticleTitleThe neighbor-joining method: a new method for reconstructing phylogenetic trees Mol Biol Evol 4 406–425

    Google Scholar 

  • M Sharma R Kambadur KG Matthews WG Somers GP Devlin JV Conaglen PJ Fowke JJ Bass (1999) ArticleTitleMyostatin, a transforming growth factor-beta superfamily member, is expressed in heart muscle and is upregulated in cardiomyocytes after infarct J Cell Physiol 180 1–9 Occurrence Handle10.1002/(SICI)1097-4652(199907)180:1<1::AID-JCP1>3.0.CO;2-V

    Article  Google Scholar 

  • M Thomas B Langley C Berry M Sharma S Kirk J Bass R Kambadur (2000) ArticleTitleMyostatin, a negative regulator of muscle growth, functions by inhibiting myoblast proliferation J Biol Chem 275 40235–40243 Occurrence Handle10.1074/jbc.M004356200

    Article  Google Scholar 

  • JD Thompson DG Higgins TJ Gibson (1994) ArticleTitleCLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position specific gap penalties and weight matrix choice Nucl Acids Res 22 4673–4680 Occurrence Handle10.1093/nar/22.22.4673

    Article  Google Scholar 

  • S Vianello L Brazzoduro VL Dalla P Belvedere L Colombo (2003) ArticleTitleMyostatin expression during development and chronic stress in zebrafish (Danio rerio) J Endocrinol 176 47–59 Occurrence Handle10.1677/joe.0.1760047

    Article  Google Scholar 

  • C Xu G Wu Y Zohar SJ Du (2003) ArticleTitleAnalysis of myostatin gene structure, expression and function in zebrafish J Exp Biol 206 IssueIDPt 22 4067–4079 Occurrence Handle10.1242/jeb.00635

    Article  Google Scholar 

  • W Yang HX Wang Y Chen Y Zhang DH Zhu (2001) ArticleTitleCloning, expression and purification of the chicken growth and differentiation factor-8 Sheng Wu Gong Cheng Xue Bao 17 460–462

    Google Scholar 

  • HQ Ye SL Chen ZX Sha MY Xu (2006) ArticleTitleDevelopment and characterization of five cell lines from heart, liver, spleen and head kidney of sea perch (Lateolabrax japonicus) J Fish Biol 69 IssueIDSuppl A 115–126 Occurrence Handle10.1111/j.1095-8649.2006.01155.x

    Article  Google Scholar 

Download references

Acknowledgments

This work was supported by grants from the National Nature Science Foundation of China (30170740) and from State 863 High-Technology R&d Project of China (2004AA626110). The authors thank Dr Xu M Y (Memorial University of Newfoundland, Canada) for technical assistance.

Author information

Authors and Affiliations

  1. Key Lab For Sustainable Utilization of Marine Fisheries Resources, Ministry of Agriculture, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao, 266071, China

    Han-Qing Ye, Song-Lin Chen, Zhen-Xia Sha & Yang Liu

Authors
  1. Han-Qing Ye
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Song-Lin Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Zhen-Xia Sha
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Yang Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Song-Lin Chen.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Ye, HQ., Chen, SL., Sha, ZX. et al. Molecular Cloning and Expression Analysis of the Myostatin Gene in Sea Perch (Lateolabrax japonicus). Mar Biotechnol 9, 262–272 (2007). https://doi.org/10.1007/s10126-006-6093-6

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10126-006-6093-6

Keywords

Search

Navigation