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.
Publication Date: 2015-06-23 PubMed ID: 26100061PubMed Central: PMC4476204DOI: 10.1186/s12917-015-0428-3Google Scholar: Lookup
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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
Researcher Affiliations
- Department of Companion Animals and Horses, University Equine Hospital, Vetmeduni Vienna, Vienna, Austria. Rene.vandenHoven@vetmeduni.ac.at.
- Jockey Club of Turkey, Istanbul, Turkey. Egur@tjk.org.
- Genomics Core Facility, VetCore, Vetmeduni Vienna, Vienna, Austria. Manuela_slamanig@gmx.at.
- Genomics Core Facility, VetCore, Vetmeduni Vienna, Vienna, Austria. Martin.Hofer@vetmeduni.ac.at.
- Department of Internal Medicine, University of Erciyes, Kayseri, Turkey. Aconmaz@erciyes.ed.tr.
- 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
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