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BMC veterinary research2015; 11; 138; doi: 10.1186/s12917-015-0428-3

Putative regulation mechanism for the MSTN gene by a CpG island generated by the SINE marker Ins227bp.

Abstract: A single nucleotide polymorphism (SNP) in the first intron of the myostatin gene (MSTN) is associated with aptness of elite Thoroughbreds to race over sprint, middle or long distances. This intronic marker (g.66493737 T ≻ C), a short interspersed nuclear element (SINE) of 227 bp (Ins227bp) insertion polymorphism in the MSTN promoter, and the adjacent SNP BIEC2-417495 have not been studied for their association with racing aptness of the average Thoroughbreds raced in countries with lower status of the racing industry. This study investigated these markers regarding their prevalence and association with performance in common race horses. Markers were genotyped by amplification refractory mutation system-quantitative PCR (ARMS-qPCR) or amplicon melting. Furthermore, we asked whether the Ins227bp marker might theoretically regulate the expression of myostatin by generating a novel target for DNA methylation or by changing binding sites for transcription factors. Putative sites for DNA methylation or binding of transcription factors were predicted by MethPrimer and by the softwares JASPAR, MatInspector and UniPROBE, respectively. Results: Pairwise linkage disequilibrium between g.66493737 T ≻ C and Ins227bp was high (r (2) = 0.93). A lower linkage was determined for g.66493737 T ≻ C and BIEC2-417495 (r (2) = 0.69) as well as for BIEC2-417495 and Ins227bp (r (2) = 0.76). The estimated frequencies for the presence of Ins227bp (I) indel and the C alleles at g.66493737 T ≻ C and BIEC2-417495 were 0.46, 0.47 and 0.43, respectively. Heterozygotes represented the most abundant genotype at each locus. The best racing distance (BRD) was significantly different between the homozygotes of each SNP (p = 0.01 to 0.03). C allele homozygotes at BIEC2-417495 or g.66493737 T ≻ C, as well as Ins227bp homozygotes earned most money on a mean distance ranging from 1211 to 1230 m. Heterozygotes earned most money on races over 1690 to 1709 m. The BRD for the T/T carriers at both SNP loci and for the SINE-free genotype was 1812 to 1854 m. Other performance parameters were not significantly different between the genotypes, except of the relative success score (RSS). The RSS was significantly slightly better on a distance of ≤ 1300 m for all carriers of the C allele and the Ins227bp compared to homozygous T genotypes and SINE-negative horses (p = 0.037 to 0.046). For distances of more than 1300 m the RSS was not significantly different between genotypes. In silico assessment indicated that the Ins227bp promoter insertion might have generated a CpG island and a few novel putative binding sites for transcription factors. Conclusions: All three target polymorphisms (Ins227bp, g.66493737 T ≻ C, BIEC2-417495) are suitable markers to assess the ability of non-elite Thoroughbreds to race at short or longer distances. The CpG island generated by Ins227bp may cause training-induced silencing of MSTN expression.
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Publication Date: 2015-06-23 PubMed ID: 26100061PubMed Central: PMC4476204DOI: 10.1186/s12917-015-0428-3Google Scholar: Lookup
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  • Non-U.S. Gov't

Summary

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This research paper explores the association of certain genetic markers with performance aptness in common racehorses. It further investigates whether one of these markers, the Ins227bp, has a potential regulatory role on the expression of myostatin.

Study Objective

The study aimed to explore the prevalence and association of certain genetic markers with racing performance in common racehorses. The researchers focused on three markers, namely, a single nucleotide polymorphism (SNP) in the first intron of the myostatin gene (MSTN), a short interspersed nuclear element (SINE) of 227 bp (Ins227bp), and the SNP BIEC2-417495.

Methodology

  • The markers were genotyped using amplification refractory mutation system-quantitative PCR (ARMS-qPCR) or amplicon melting.
  • Pairwise linkage disequilibrium between the markers was measured to determine their co-occurrence.
  • The study attempted to predict whether the Ins227bp could regulate myostatin expression by generating a target for DNA methylation or by changing the binding sites for transcription factors. This was done using software like MethPrimer, JASPAR, MatInspector, and UniPROBE.

Results

  • High linkage disequilibrium was observed between the SNP in the first intron of the MSTN gene and Ins227bp (r(2) = 0.93).
  • The observed frequencies for the presence of Ins227bp, C alleles at the SNP in MSTN, and BIEC2-417495 were 0.46, 0.47, and 0.43, respectively.
  • The best racing distance (BRD) showed significant differences among homozygote horses for each SNP.
  • C allele homozygotes at BIEC2-417495 or the SNP in MSTN, along with Ins227bp homozygotes, earned most money at mean distances ranging from 1211 to 1230 m.
  • The assessment indicated that the Ins227bp promoter insertion might have generated a CpG island and a few novel putative binding sites for transcription factors.
  • Other performance parameters were not significantly different between the genotypes, except for the relative success score (RSS). The RSS was noticeably better at a distance of ≤ 1300 m for all carriers of the C allele and the Ins227bp than their T genotype and SINE-negative counterparts.

Conclusions

  • The three target polymorphisms (Ins227bp, the SNP in MSTN, BIEC2-417495) are suitable markers to assess the racing performance of non-elite Thoroughbreds at different distances.
  • The CpG island that potentially resulted from Ins227bp may cause training-induced silencing of MSTN expression. This could have important implications for the regulation of myostatin, a gene known to inhibit muscle growth, ultimately affecting the horse’s performance.

Cite This Article

APA
van den Hoven R, Gür E, Schlamanig M, Hofer M, Onmaz AC, Steinborn R. (2015). Putative regulation mechanism for the MSTN gene by a CpG island generated by the SINE marker Ins227bp. BMC Vet Res, 11, 138. https://doi.org/10.1186/s12917-015-0428-3

Publication

BMC veterinary research
ISSN: 1746-6148
NlmUniqueID: 101249759
Country: England
Language: English
Volume: 11
Pages: 138
PII: 138

Researcher Affiliations

van den Hoven, René
  • Department of Companion Animals and Horses, University Equine Hospital, Vetmeduni Vienna, Vienna, Austria. Rene.vandenHoven@vetmeduni.ac.at.
Gür, Emre
  • Jockey Club of Turkey, Istanbul, Turkey. Egur@tjk.org.
Schlamanig, Manuela
  • Genomics Core Facility, VetCore, Vetmeduni Vienna, Vienna, Austria. Manuela_slamanig@gmx.at.
Hofer, Martin
  • Genomics Core Facility, VetCore, Vetmeduni Vienna, Vienna, Austria. Martin.Hofer@vetmeduni.ac.at.
Onmaz, Ali Cesur
  • Department of Internal Medicine, University of Erciyes, Kayseri, Turkey. Aconmaz@erciyes.ed.tr.
Steinborn, Ralf
  • Genomics Core Facility, VetCore, Vetmeduni Vienna, Vienna, Austria. Ralf.Steinborn@vetmeduni.ac.at.

MeSH Terms

  • Alleles
  • Animals
  • CpG Islands / genetics
  • Gene Expression Regulation
  • Genetic Markers
  • Genotype
  • Horses / genetics
  • Horses / metabolism
  • Linkage Disequilibrium
  • Myostatin / genetics
  • Myostatin / metabolism
  • Polymorphism, Single Nucleotide
  • Short Interspersed Nucleotide Elements / genetics

References

This article includes 33 references
  1. Hill EW, Gu J, Eivers SS, Fonseca RG, McGivney BA, Govindarajan P, Orr N, Katz LM, MacHugh DE. A sequence polymorphism in MSTN predicts sprinting ability and racing stamina in thoroughbred horses.. PLoS One 2010 Jan 20;5(1):e8645.
    doi: 10.1371/journal.pone.0008645pmc: PMC2808334pubmed: 20098749google scholar: lookup
  2. Hill EW, McGivney BA, Gu J, Whiston R, Machugh DE. A genome-wide SNP-association study confirms a sequence variant (g.66493737C>T) in the equine myostatin (MSTN) gene as the most powerful predictor of optimum racing distance for Thoroughbred racehorses.. BMC Genomics 2010 Oct 11;11:552.
    doi: 10.1186/1471-2164-11-552pmc: PMC3091701pubmed: 20932346google scholar: lookup
  3. Tozaki T, Miyake T, Kakoi H, Gawahara H, Sugita S, Hasegawa T, Ishida N, Hirota K, Nakano Y. A genome-wide association study for racing performances in Thoroughbreds clarifies a candidate region near the MSTN gene.. Anim Genet 2010 Dec;41 Suppl 2:28-35.
    doi: 10.1111/j.1365-2052.2010.02095.xpubmed: 21070273google scholar: lookup
  4. Tozaki T, Hill EW, Hirota K, Kakoi H, Gawahara H, Miyake T, Sugita S, Hasegawa T, Ishida N, Nakano Y, Kurosawa M. A cohort study of racing performance in Japanese Thoroughbred racehorses using genome information on ECA18.. Anim Genet 2012 Feb;43(1):42-52.
    doi: 10.1111/j.1365-2052.2011.02201.xpubmed: 22221024google scholar: lookup
  5. Binns MM, Boehler DA, Lambert DH. Identification of the myostatin locus (MSTN) as having a major effect on optimum racing distance in the Thoroughbred horse in the USA.. Anim Genet 2010 Dec;41 Suppl 2:154-8.
    doi: 10.1111/j.1365-2052.2010.02126.xpubmed: 21070290google scholar: lookup
  6. Tozaki T, Sato F, Hill EW, Miyake T, Endo Y, Kakoi H, Gawahara H, Hirota K, Nakano Y, Nambo Y, Kurosawa M. Sequence variants at the myostatin gene locus influence the body composition of Thoroughbred horses.. J Vet Med Sci 2011 Dec;73(12):1617-24.
    doi: 10.1292/jvms.11-0295pubmed: 21836385google scholar: lookup
  7. McPherron AC, Lawler AM, Lee SJ. Regulation of skeletal muscle mass in mice by a new TGF-beta superfamily member.. Nature 1997 May 1;387(6628):83-90.
    doi: 10.1038/387083a0pubmed: 9139826google scholar: lookup
  8. McPherron AC, Lee SJ. Double muscling in cattle due to mutations in the myostatin gene.. Proc Natl Acad Sci U S A 1997 Nov 11;94(23):12457-61.
    doi: 10.1073/pnas.94.23.12457pmc: PMC24998pubmed: 9356471google scholar: lookup
  9. Grobet L, Poncelet D, Royo LJ, Brouwers B, Pirottin D, Michaux C, Ménissier F, Zanotti M, Dunner S, Georges M. Molecular definition of an allelic series of mutations disrupting the myostatin function and causing double-muscling in cattle.. Mamm Genome 1998 Mar;9(3):210-3.
    doi: 10.1007/s003359900727pubmed: 9501304google scholar: lookup
  10. Mosher DS, Quignon P, Bustamante CD, Sutter NB, Mellersh CS, Parker HG, Ostrander EA. A mutation in the myostatin gene increases muscle mass and enhances racing performance in heterozygote dogs.. PLoS Genet 2007 May 25;3(5):e79.
    doi: 10.1371/journal.pgen.0030079pmc: PMC1877876pubmed: 17530926google scholar: lookup
  11. Petersen JL, Valberg SJ, Mickelson JR, McCue ME. Haplotype diversity in the equine myostatin gene with focus on variants associated with race distance propensity and muscle fiber type proportions.. Anim Genet 2014 Dec;45(6):827-35.
    doi: 10.1111/age.12205pmc: PMC4211974pubmed: 25160752google scholar: lookup
  12. Nielsen BD, Turner KK, Ventura BA, Woodward AD, O'Connor CI. Racing speeds of quarter horses, thoroughbreds and Arabians.. Equine Vet J Suppl 2006 Aug;(36):128-32.
    doi: 10.1111/j.2042-3306.2006.tb05528.xpubmed: 17402407google scholar: lookup
  13. McMiken DF. An energetic basis of equine performance.. Equine Vet J 1983 Apr;15(2):123-33.
    doi: 10.1111/j.2042-3306.1983.tb01734.xpubmed: 6873045google scholar: lookup
  14. Bower MA, McGivney BA, Campana MG, Gu J, Andersson LS, Barrett E, Davis CR, Mikko S, Stock F, Voronkova V, Bradley DG, Fahey AG, Lindgren G, MacHugh DE, Sulimova G, Hill EW. The genetic origin and history of speed in the Thoroughbred racehorse.. Nat Commun 2012 Jan 24;3:643.
    doi: 10.1038/ncomms1644pubmed: 22273681google scholar: lookup
  15. Petersen JL, Mickelson JR, Rendahl AK, Valberg SJ, Andersson LS, Axelsson J, Bailey E, Bannasch D, Binns MM, Borges AS, Brama P, da Câmara Machado A, Capomaccio S, Cappelli K, Cothran EG, Distl O, Fox-Clipsham L, Graves KT, Guérin G, Haase B, Hasegawa T, Hemmann K, Hill EW, Leeb T, Lindgren G, Lohi H, Lopes MS, McGivney BA, Mikko S, Orr N, Penedo MC, Piercy RJ, Raekallio M, Rieder S, Røed KH, Swinburne J, Tozaki T, Vaudin M, Wade CM, McCue ME. Genome-wide analysis reveals selection for important traits in domestic horse breeds.. PLoS Genet 2013;9(1):e1003211.
    doi: 10.1371/journal.pgen.1003211pmc: PMC3547851pubmed: 23349635google scholar: lookup
  16. SanGiacomo NE. The Impact of Myostatin Genetic Polymorphism on Muscle Conformation in the Horse. PhD Thesis, Cornell University, College of Agriculture and Life Sciences, Animal Science; 2013.
  17. McGivney BA, Browne JA, Fonseca RG, Katz LM, Machugh DE, Whiston R, Hill EW. MSTN genotypes in Thoroughbred horses influence skeletal muscle gene expression and racetrack performance.. Anim Genet 2012 Dec;43(6):810-2.
    doi: 10.1111/j.1365-2052.2012.02329.xpubmed: 22497477google scholar: lookup
  18. Bianchi M, Crinelli R, Giacomini E, Carloni E, Radici L, Magnani M. Yin Yang 1 intronic binding sequences and splicing elicit intron-mediated enhancement of ubiquitin C gene expression.. PLoS One 2013;8(6):e65932.
    doi: 10.1371/journal.pone.0065932pmc: PMC3680475pubmed: 23776572google scholar: lookup
  19. Parra G, Bradnam K, Rose AB, Korf I. Comparative and functional analysis of intron-mediated enhancement signals reveals conserved features among plants.. Nucleic Acids Res 2011 Jul;39(13):5328-37.
    doi: 10.1093/nar/gkr043pmc: PMC3141229pubmed: 21427088google scholar: lookup
  20. Park SG, Hannenhalli S, Choi SS. Conservation in first introns is positively associated with the number of exons within genes and the presence of regulatory epigenetic signals.. BMC Genomics 2014 Jun 26;15(1):526.
    doi: 10.1186/1471-2164-15-526pmc: PMC4085337pubmed: 24964727google scholar: lookup
  21. Guenther CA, Tasic B, Luo L, Bedell MA, Kingsley DM. A molecular basis for classic blond hair color in Europeans.. Nat Genet 2014 Jul;46(7):748-52.
    doi: 10.1038/ng.2991pmc: PMC4704868pubmed: 24880339google scholar: lookup
  22. Palmer AA, Dulawa SC. Murine Warriors or Worriers: The Saga of Comt1, B2 SINE Elements, and the Future of Translational Genetics.. Front Neurosci 2010;4:177.
    doi: 10.3389/fnins.2010.00177pmc: PMC2958038pubmed: 20976038google scholar: lookup
  23. de Souza FS, Franchini LF, Rubinstein M. Exaptation of transposable elements into novel cis-regulatory elements: is the evidence always strong?. Mol Biol Evol 2013 Jun;30(6):1239-51.
    doi: 10.1093/molbev/mst045pmc: PMC3649676pubmed: 23486611google scholar: lookup
  24. Arnaud P, Goubely C, Pélissier T, Deragon JM. SINE retroposons can be used in vivo as nucleation centers for de novo methylation.. Mol Cell Biol 2000 May;20(10):3434-41.
    doi: 10.1128/MCB.20.10.3434-3441.2000pmc: PMC85636pubmed: 10779333google scholar: lookup
  25. Steinborn R, Schinogl P, Zakhartchenko V, Achmann R, Schernthaner W, Stojkovic M, Wolf E, Müller M, Brem G. Mitochondrial DNA heteroplasmy in cloned cattle produced by fetal and adult cell cloning.. Nat Genet 2000 Jul;25(3):255-7.
    doi: 10.1038/77000pubmed: 10888867google scholar: lookup
  26. Zuker M. Mfold web server for nucleic acid folding and hybridization prediction.. Nucleic Acids Res 2003 Jul 1;31(13):3406-15.
    doi: 10.1093/nar/gkg595pmc: PMC169194pubmed: 12824337google scholar: lookup
  27. Barrett JC, Fry B, Maller J, Daly MJ. Haploview: analysis and visualization of LD and haplotype maps.. Bioinformatics 2005 Jan 15;21(2):263-5.
    doi: 10.1093/bioinformatics/bth457pubmed: 15297300google scholar: lookup
  28. Sandelin A, Alkema W, Engström P, Wasserman WW, Lenhard B. JASPAR: an open-access database for eukaryotic transcription factor binding profiles.. Nucleic Acids Res 2004 Jan 1;32(Database issue):D91-4.
    doi: 10.1093/nar/gkh012pmc: PMC308747pubmed: 14681366google scholar: lookup
  29. Mathelier A, Zhao X, Zhang AW, Parcy F, Worsley-Hunt R, Arenillas DJ, Buchman S, Chen CY, Chou A, Ienasescu H, Lim J, Shyr C, Tan G, Zhou M, Lenhard B, Sandelin A, Wasserman WW. JASPAR 2014: an extensively expanded and updated open-access database of transcription factor binding profiles.. Nucleic Acids Res 2014 Jan;42(Database issue):D142-7.
    pmc: PMC3965086pubmed: 24194598doi: 10.1093/nar/gkt997google scholar: lookup
  30. Quandt K, Frech K, Karas H, Wingender E, Werner T. MatInd and MatInspector: new fast and versatile tools for detection of consensus matches in nucleotide sequence data.. Nucleic Acids Res 1995 Dec 11;23(23):4878-84.
    doi: 10.1093/nar/23.23.4878pmc: PMC307478pubmed: 8532532google scholar: lookup
  31. Hume MA, Barrera LA, Gisselbrecht SS, Bulyk ML. UniPROBE, update 2015: new tools and content for the online database of protein-binding microarray data on protein-DNA interactions.. Nucleic Acids Res 2015 Jan;43(Database issue):D117-22.
    doi: 10.1093/nar/gk၅pmc: PMC4383892pubmed: 25378322google scholar: lookup
  32. Li LC, Dahiya R. MethPrimer: designing primers for methylation PCRs.. Bioinformatics 2002 Nov;18(11):1427-31.
    doi: 10.1093/bioinformatics/18.11.1427pubmed: 12424112google scholar: lookup
  33. Wright S. Coefficients of Inbreeding and Relationship. Am Nat 1922;56:330–338.
    doi: 10.1086/279872google scholar: lookup

Citations

This article has been cited 7 times.
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