Polymorphism Of Myostatin Gene Research Articles

Gene Expression and Polymorphism of Myostatin Gene and its Association with Growth Traits in Chicken

ABSTRACTMyostatin is a member of TGF-β super family and is directly involved in regulation of body growth through limiting muscular growth. A study was carried out in three chicken lines to identify the polymorphism in the coding region of the myostatin gene through SSCP and DNA sequencing. A total of 12 haplotypes were observed in myostatin coding region of chicken. Significant associations between haplogroups with body weight at day 1, 14, 28, and 42 days, and carcass traits at 42 days were observed across the lines. It is concluded that the coding region of myostatin gene was polymorphic, with varied levels of expression among lines and had significant effects on growth traits. The expression of MSTN gene varied during embryonic and post hatch development stage.

Association between myostatin gene (MSTN-1) polymorphism and growth traits in domesticated rainbow trout (Oncorhynchus mykiss)

In this study, the polymorphism of myostatin gene (MSTN-1) as a candidate gene for quantitative traits was analyzed in the rainbow trout (Oncorhynchus mykiss). The association between the MSTN-1 polymorphisms and growth traits were also studied. Forty broodstock parents of rainbow trout from two farms including Yasuj and Shahrkord were selected and tagged. These spawning were mated and then juveniles were raised at the ponds until 5months post-hatching. Two single nucleotide polymorphisms (SNPs: g.660T>C and g.1904C>A) in intron 1 and 2 regions of the MSTN-1 gene in rainbow trout were determined by DNA sequencing and PCR-single stranded conformation polymorphism (PCR-SSCP) methods. The assessment results demonstrated that these two site mutations are synonymous mutations and therefore 6 haplotypes were revealed in these samples (n=120). Moreover, One way ANOVA of the SNPs and growth traits revealed the genotypes of SNP g.1904C>A are associated with the length and body weight at 90 and 150days of age (P<0.05), whereas no significant association was found between the SNP g.660T>C and growth trait parameters. These results provide evidence that SNPs in MSTN-1 gene may be associated with length and body weight during the early stages of life (from hatching to 5months of age), therefore MSTN-1 gene may be used for rainbow trout breeding program.

Characterization of &amp;lt;i&amp;gt;Myostatin&amp;lt;/i&amp;gt; Gene in Nile Tilapia (&amp;lt;i&amp;gt;Oreochromis niloticus&amp;lt;/i&amp;gt;), the Possible Association of &amp;lt;i&amp;gt;BsmI-exon 2&amp;lt;/i&amp;gt; Polymorphism with Its Growth

Myostatin (MSTN) gene is a negative regulator of skeletal muscle. In this study, we try to evaluate MSTN as a candidate gene for a marker assisted selection in Nile tilapia growth trait. Molecular characterization of MSTN gene was done using seven designed primers. We demonstrated that, O. niloticus MSTN sequence and its promoter is as for all known vertebrates. Novel SNPs were identified in coding and non-coding regions compared with MSTN gene of O. niloticus x O. aureus hybrid, three non-synonymous SNPs were found at MSTN coding region; two at exon one 369 C>A, 831 T>A, and one at exon 3 2637 G>A, altering Thr 38-Pro, Glu 121-Val and Tyr 375-Cys respectively. Phylogenetic analysis revealed high similarity (99.2) with MSTN gene of the hybrid with O. aurous. BsmI induced cutting pattern at MSTN-exon 2 (607-bp). Among two hundred monosex male fish, two different genotypes were reported; AB genotype (607-bp, 507-bp and 100-bp) and BB genotype (507-bp and 100-bp fragments), were produced. Most large-size fish are included in AB genotype with 0.8 frequency and significantly increased body weight compared with small size fish, which are mostly included in BB genotype with 0.9 frequencies. Novel BsmI-exon 2 polymorphism of MSTN gene can be used as a marker assisted selection for large body weight in heterozygous Nile tilapia fish.

Myostatin Gene Polymorphism in an Elderly Sarcopenic Turkish Population.

One of the genetic contributors to sarcopenia predisposition is Myostatin (MSTN), which in humans encodes myostatin, a 376 amino acid growth factor protein that negatively regulates muscle growth. The aim of this study was to investigate MSTN polymorphisms in an elderly sarcopenic population in Turkey and determine how they relate to sarcopenia. The study included nursing home residents who were aged ≥65 years. Sarcopenia screening was performed using "The European Working Group on Sarcopenia in Older People" guidelines. Blood sample was taken from each participant and DNA was obtained from the peripheral blood. MSTN polymorphisms were genotyped by polymerase chain reaction and restriction fragment length polymorphism methods. A total of 152 elderly patients were included in the study. The rate of sarcopenia was determined to be 41.4%. The DNA nucleotide sequence of all three MSTN exons was determined for each study participant. Among the 152 patients, only 6 (3.9%) showed an MSTN K153R heterozygous mutation. Among these, three participants were sarcopenic and three were nonsarcopenic. No statistically significant difference in the polymorphism frequency between the sarcopenic and control groups was observed (p=0.664). MSTN genotyping revealed that only 3.9% (6/152) of participants had the MSTN K153R heterozygous mutation. Despite the detection of this mutation in the study group, no relationship was found between this mutation and sarcopenia.

Analysis of the 227 bp short interspersed nuclear element (SINE) insertion of the promoter of the myostatin (MSTN) gene in different horse breeds.

The myostatin (MSTN) gene encodes a protein known to be a negative regulator of muscle mass in mammalian species. Different polymorphisms of the horse (Equus caballus) MSTN gene have been identified, including single nucleotide polymorphisms and a short interspersed nuclear element (SINE) insertion of 227 bp within the promoter of the gene. The SINE insertion has been associated with performance traits in Thoroughbred racehorses and it was proposed as a predictor of optimum racing distance. The aims of this study were to perform in silico analysis to identify putative gains or abrogation of transcription-factor binding sites (TFBSs) generated by the SINE allele of the promoter and to analyse the frequency of the SINE insertion in horses used for racing (gallop and trot) and other purposes. The SINE insertion was genotyped in 227 horses from 10 breeds belonging to different morphological types (brachimorphic, mesomorphic, meso-dolichomorphic and dolichomorphic). The presence of the insertion was confirmed in the Quarter Horse (SINE allele frequency of 0.81) and in the Thoroughbred (0.51), whereas the SINE allele did not segregate in any of the other analysed breeds. As the SINE MSTN gene polymorphism may be population or breed specific, it is not a useful marker for association studies in all breeds.

Rabbit MSTN gene polymorphisms and genetic effect analysis.

We analyzed meat samples of nine pure lines of rabbit and its 37 hybrid combinations by sequencing and single-strand conformation polymorphism techniques to explore genetic polymorphisms of all the three exon regions and part of the 5'-regulatory region of the myostatin (MSTN) gene. Thus, we detected a single nucleotide mutation (T→C) on the 476 locus of the 5'-regulatory region, but no mutation sites were detected in the exon areas. The correlation analysis showed that the mutation had some favorable genetic effects, and it resulted in increased liver weight, carcass weight, forelegs weight, back and waist weight, ham weight, and tare weight, whereas it decreased muscle drip loss and cooking loss (P < 0.05). These results suggest that the mutations in the upstream regulatory region of the MSTN gene are beneficial to the rabbit soma development, and the mutations can be used as molecular markers for the selection of the meat quality of rabbits.

Single nucleotide polymorphisms of myostatin gene in Chinese domestic horses

The myostatin gene (MSTN) is a genetic determinant of skeletal muscle growth. Single nucleotide polymorphisms (SNP) in MSTN are of importance due to their strong associations with horse racing performances. In this study, we screened the SNPs in MSTN gene in 514 horses from 15 Chinese horse breeds. Six SNPs (g.26T>C, g.156T>C, g.587A>G, g.598C>T, g.1485C>T, g.2115A>G) in MSTN gene were detected by sequencing and genotyped using PCR-RFLP method. The g.587A>G and g.598C>T residing in the 5′UTR region were novel SNPs identified by this study. The g.2115A>G which have previously been associated with racing performances were present in Chinese horse breeds, providing valuable genetic information for evaluating the potential racing performances in Chinese domestic breeds. The six SNPs together defined thirteen haplotypes, demonstrating abundant haplotype diversities in Chinese horses. Most of the haplotypes were shared among different breeds with no haplotype restricted to a specific region or a single horse breed. AMOVA analysis indicated that most of the genetic variance was attributable to differences among individuals without any significant contribution by the four geographical groups. This study will provide fundamental and instrumental genetic information for evaluating the potential racing performances of Chinese horse breeds.

Rapid Genotyping of MSTN Gene Polymorphism Using High-resolution Melting for Association Study in Rabbits.

The myostatin (MSTN) gene, as a negative regulator of skeletal muscle growth, has been proposed to be associated with production traits in farm animals. In the present study, a T/C variant at −125 bp (relative to ATG start codon) of 5’regulatory region of rabbit MSTN was identified by direct sequencing. Two hundred and twenty two rabbits, which were randomly sampled from 3 breeds (Ira rabbits, Champagne rabbits and Tianfu black rabbits), were genotyped by high-resolution melting (HRM). Comparing the genotyping results of 47 samples with direct sequencing, the HRM showed high sensitivity (0.96) and high specificity (0.98). In the three rabbit breeds, the allele C was the predominant allele. The polymorphic site showed high heterozygosity (He = 0.48) and high effective number of alleles (Ne = 1.91). The genetic diversity was reasonably informative (0.25<PIC<0.50). The association analysis showed that the genotype TC had significant effect on the 84-d-weight of rabbits compared with genotype CC (p = 0.047). In contrast, the genotypes had no significant effect on other production traits. These results showed that HRM could be effectively used for genotyping analysis of MSTN gene. The T/C variant in 5’regulatory region of MSTN might be one of the candidate SNP loci affecting the trait of 84-d-weight.

Association of the Myostatin gene with obesity, abdominal obesity and low lean body mass and in non-diabetic Asian Indians in north India.

BackgroundTo determine the association of the A55T and K153R polymorphisms of the Myostatin gene with obesity, abdominal obesity and lean body mass (LBM) in Asian Indians in north India.Materials and MethodsA total of 335 subjects (238 men and 97 women) were assessed for anthropometry, % body fat (BF), LBM and biochemical parameters. Associations of Myostatin gene polymorphisms were evaluated with anthropometric, body composition and biochemical parameters. In A55T polymorphism, BMI (p = 0.04), suprailiac skinfold (p = 0.05), total skinfold (p = 0.008), %BF (p = 0.002) and total fat mass (p = 0.003) were highest and % LBM (p = 0.03) and total LBM (Kg) were lowest (p = 0.04) in subjects with Thr/Thr genotype as compared to other genotypes. Association analysis of K153R polymorphism showed that subjects with R/R genotype had significantly higher BMI (p = 0.05), waist circumference (p = 0.04), %BF (p = 0.04) and total fat mass (p = 0.03), and lower %LBM (p = 0.02) and total LBM [(Kg), (p = 0.04)] as compared to other genotypes. Using a multivariate logistic regression model after adjusting for age and sex, subjects with Thr/Thr genotype of A55T showed high risk for high %BF (OR, 3.92, 95% Cl: 2.61–12.41), truncal subcutaneous adiposity (OR, 2.9, 95% Cl: 1.57–6.60)] and low LBM (OR, 0.64, 95% CI: 0.33–0.89) whereas R/R genotype of K153R showed high risk of obesity (BMI; OR, 3.2, 95% CI: 1.2–12.9; %BF, OR, 3.6, 95% CI: 1.04–12.4), abdominal obesity (OR, 2.12, 95% CI: 2.71–14.23) and low LBM (OR, 0.61, 95% CI: 0.29–0.79).Conclusions/SignificanceWe report that variants of Myostatin gene predispose to obesity, abdominal obesity and low lean body mass in Asian Indians in north India.

Polymorphism of Myostatin Gene in Intron 1 and 2 and Exon 3, and Their Associations with Yearling Weight, Using PCR-RFLP and PCR-SSCP Techniques in Zel Sheep

The aim of present study was to investigate myostatin gene polymorphism and its association with yearling weight records in Zel sheep using PCR-RFLP and PCR-SSCP methods. Blood samples were collected from 200 Zel sheep, randomly, and DNA was extracted using modified salting out method. Polymerase chain reaction was carried out to amplify 337, 222, and 311 bp fragments, respectively, comprising a part of exon 3, intron 1, and intron 2 of myostatin gene. In addition, exon 3 was digested by HaeIII enzyme under RFLP method, and introns 1 and 2 were studied using SSCP. Under RFLP method, all samples showed mm genotype. Under SSCP method, intron 1 was also monomorph but intron 2 was polymorph (AA, AB, and BB). The allelic frequencies for A and B were 75.5 and 24.5%, respectively. This locus was not in Hardy-Weinberg equilibrium (P < 0.05), and there was no significant effect of myostatin gene on yearling weights.

Contribution of Myostatin gene polymorphisms to normal variation in lean mass, fat mass and peak BMD in Chinese male offspring

Myostatin gene is a member of the transforming growth factor-β (TGF-β) family that negatively regulates skeletal muscle growth. Genetic polymorphisms in Myostatin were found to be associated with the peak bone mineral density (BMD) in Chinese women. The purpose of this study was to investigate whether myostatin played a role in the normal variation in peak BMD, lean mass (LM), and fat mass (FM) of Chinese men. Four hundred male-offspring nuclear families of Chinese Han ethnic group were recruited. Anthropometric measurements, including the peak BMD, body LM and FM were measured using dual-energy X-ray absorptiometry (DXA). The single nucleotide polymorphisms (SNPs) studied were tag-SNPs selected by sequencing. Both rs2293284 and +2278GA were genotyped using TaqMan assay, and rs3791783 was genotyped with PCR-restriction fragment length polymorphism (RFLP) analysis. The associations of the SNPs with anthropometric variations were analyzed using the quantitative transmission disequilibrium test (QTDT). Using QTDT to detect within-family associations, neither single SNP nor haplotype was found to be associated with peak BMD at any bone site. However, rs3791783 was found to be significantly associated with fat mass of the trunk (P<0.001). Moreover, for within-family associations, haplotypes AGG, AAA, and TGG were found to be significantly associated with the trunk fat mass (all P<0.001). Our results suggest that genetic variation within myostatin may play a role in regulating the variation in fat mass in Chinese males. Additionally, the myostatin gene may be a candidate that determines body fat mass in Chinese men.

Association of myostatin gene polymorphisms with obesity in Chinese north Han human subjects

ContextMyostatin (MSTN) is a member of the TGF-β superfamily of signal transduction proteins, which plays an important role in muscular growth and lipid metabolism. ObjectiveTo study the association of myostatin gene polymorphisms with obesity in Chinese north Han human subjects. Design297 healthy and 606 over-weight/obesity Chinese north Han subjects were selected as healthy control group and overweight/obesity group, respectively. The methods of DNA Sequencing, Restriction Fragment Length Polymorphism (RFLP) and TaqMan® probe were used to screen myostatin gene SNPs and clarify genotype in every individual. ResultsTotal 11 SNPs in MSTN gene were identified by DNA sequencing and three SNPs including rs35781413 (G/A), rs3791783 (A/G) and rs3791782 (A/G) were selected for further study in total 903 samples. The results showed that the frequency of AA genotype of rs3791783 A/G SNP was significantly higher (56.4% vs. 50.8%) and the frequency GG genotype was significantly lower (3.2% vs. 6.7%) in overweight/obese patients than in normal weight subjects. A logistic regression analysis under a recessive inheritance model (AA+AG vs.GG) demonstrated that the Odd ratio for AA+AG vs.GG were 1.985 (95% CI 1.078–3.643; P=0.029). Among three genotypes of rs3791783, the subjects with AA genotype have much more higher body weight, BMI, waist circumference, TC, TG and LDL-C than those with GG genotype. ConclusionsOur data firstly suggest that genetic variant rs3791783 A/G in myostatin gene are associated with obesity. The A allele carriers in rs3791783 SNP have an increased susceptibility to obesity compared with the G allele carriers. Participants with AA genotype in rs3791783 SNP site will have higher risk suffered from overweight or obesity than those with GG genotype.

IDENTIFICATION OF MYOSTATIN GENE c.960delG LOCUS POLYMORPHISM IN INDONESIAN LOCAL SHEEP BY USING PCR-SSCP METHOD

Myostatin (MSTN) is a protein inhibit muscle growth. This protein is a member of a superfamily ofmolecules called transforming growth factors beta (TGF-b). Deletion in c.960delG (1-bp deletion atposition 960) disrupts the reading frame from amino acid (aa) position 320 to ending in a premature stopcodon in aa position 359 have been found in Norwegian White Sheep. This deletion in the myostatingene is responsible to increase muscle mass, also known as 'double muscling', in sheep. The purpose ofthis study was to identify the polymorphism of myostatin gene in c.960delG locus of local sheep inIndonesia. The 832 DNA sampels from sheep were collected from 13 populations belonging to thePriangan (86), Javanese Thin Tail (389 i.e. Jonggol, Banjar, Ciomas), Javanese Fat Tail (94), West NusaTenggara (136), Rote Island/East Nusa Tenggara (35), Kisar Island/Southwest Maluku (22),Donggala/South East Celebes (45) and Batur cross breed/Wonosobo, Central Java (25). A gene fragmentof MSTN c.960delG length 299 bp was successfully amplified by using the technique of PCR(polymerase chain reaction) and genotyped by SSCP (single strand conformation polymorphism). Theresult showed no polymorphism in this gene. All sheep tested had G/G genotype for c.960delG locus.

Polymorphisms of myostatin gene (MSTN) in four goat breeds and their effects on Boer goat growth performance

Polymorphisms of myostatin (MSTN) gene were investigated as a candidate marker for goat growth in 687 individuals by gene sequencing and polymerase chain reaction restriction fragment length polymorphism methods. Three potential genotypes (AA, AB and BB) of 5 bp indel (1,256 TTTTA/-) in 5'UTR were detected in four breeds. The polymorphism (CC, CD and DD) of substitution (1,388 T/A) in exon 1 was only segregating in Boer. Genotype AB resulted in significant increases in body weights at birth (BW0), 90 (BW90) and 300 (BW300) days of age, and birth body length compared to BB (P < 0.05). Whilst genotype CD contributed to heavier BW0 and BW90, and larger birth body height (BH) compared to CC (P < 0.05). Individuals either with AB or CD genotype also had greater values in other body sizes, although no significant differences appeared (P > 0.05). When in combination, the combined genotype ABCD displayed positive impacts on better growth traits in BW0, BW90, BW300, BH and BCG (P < 0.05). These results indicate that these markers in myostatin (MSTN) are associated with Boer growth and may be useful for marker assisted selection.

Analysis of the Single-Nucleotide Polymorphism in the 5′UTR and Part of Intron I of the Sheep MSTN Gene

The myostatin (MSTN) gene region encompassing the 5'UTR and part of intron I was sequenced in animals of two herds of Latvian Darkhead sheep to extend data on the ovine MSTN gene polymorphism and to provide information useful for local breed conservation. Two and four polymorphic loci were revealed in the 5'UTR and intron I. Four and five local haplotypes were constructed, respectively. The genotyping data obtained and that previously reported for the same genomic region were combined in one dataset for the haplotype analysis. Recombination events were detected between loci (c.-40, c.-37) in the 5'UTR and (c.373+18, c.373+101) and (c.373+101, c.373+241) in intron I. Single-nucleotide polymorphisms at c.373+249 and c.373+323 appear to be involved in the strong linkage (p < 0.01). Linkage blocks (c.373+241, c.373+243) and (c.373+241, c.373+259) were revealed at nominal (p < 0.05) level of probability. Haplotype-specific patterns of the transcription factor binding sites predicted in silico were constructed to evaluate a putative functional significance of the particular alleles and haplotypes. A nucleotide at c.373+18 was shown to influence the pre-mRNA secondary structure. DNA curvature predicted in silico for allele c.373+101C was proven experimentally. A possible impact of the particular polymorphisms on the transcription and/or splicing efficiency is discussed.

Contribution of the Myostatin gene polymorphisms to normal variation in lean mass and fat mass in Chinese male offspring nuclear families

Contribution of the Myostatin gene polymorphisms to normal variation in lean mass and fat mass in Chinese male offspring nuclear families

Association between myostatin gene polymorphisms and peak BMD variation in Chinese nuclear families

We identified 17 polymorphisms in myostatin by sequencing, and three informative single nucleotide polymorphisms (SNPs) were selected for further observation for their association with peak BMD of women in 401 Chinese nuclear families. Our results suggest that genetic polymorphisms in myostatin likely play a role in attainment of peak BMD in Chinese women. Myostatin is a TGF-beta family member that is a negative regulator of skeletal muscle growth. We identified SNPs in myostatin by direct sequencing. Furthermore, using a quantitative transmission disequilibrium test (QTDT). we tested and further test whether SNPs were associated with peak bone mineral density (BMD) variation at the spines and hips of 401 Chinese nuclear families. We identified 17 polymorphisms in myostatin by sequencing. Next, we selected three informative SNPs for further observation of an association with peak BMD of premenopausal women in 401 Chinese nuclear families. Using QTDT for the within-family association, we found significant association between rs2293284 and total hip, femoral neck, and trochanter BMD (all p < 0.05), while rs7570532 was associated with total hip and trochanter BMD (p = 0.034 and p = 0.035, respectively). The within-family association was significant between BMI and +2278G > A (p = 0.022). Subsequent permutations were in agreement with these significant within-family association results. Moreover, analyses of the haplotypes confer further evidence for association of rs2293284 and rs7570532 with hip peak BMD variation. These results suggest, for the first time, the genetic polymorphisms in myostatin likely play a role in attainment of peak BMD in Chinese women.

Associations of myostatin gene polymorphisms with performance and mortality traits in broiler chickens

Myostatin is a negative regulator of skeletal muscle growth. We evaluated effects of myostatin polymorphisms in three elite commercial broiler chicken lines on mortality, growth, feed conversion efficiency, ultrasound breast depth, breast percentage, eviscerated carcass weight, leg defects, blood oxygen level, and hen antibody titer to infectious bursal disease virus vaccine. Progeny mean data adjusted for fixed and mate effects and DNA from 100 sires per line were used. Single nucleotide polymorphisms (SNPs) of the myostatin gene segregating in these lines were identified by designing specific primers, amplifying individual DNA in each line by polymerase chain reaction, cloning, sequencing and aligning the corresponding products. Individual sires were genotyped for five identified SNPs which contributed to eight haplotypes. Frequencies of SNP alleles and haplotypes differed between lines. Using the allele substitution effect model, the myostatin SNPs were found to have significant (P < 0.031) associations with growth, mortality, blood oxygen and hen antibody titer to infectious bursal disease virus vaccine, although the associations were not often consistent across lines. These results suggest that the myostatin gene has pleiotropic effects on broiler performance.

Associations of myostatin gene polymorphisms with performance and mortality traits in broiler chickens

Myostatin is a negative regulator of skeletal muscle growth. We evaluated effects of myostatin polymorphisms in three elite commercial broiler chicken lines on mortality, growth, feed conversion efficiency, ultrasound breast depth, breast percentage, eviscerated carcass weight, leg defects, blood oxygen level, and hen antibody titer to infectious bursal disease virus vaccine. Progeny mean data adjusted for fixed and mate effects and DNA from 100 sires per line were used. Single nucleotide polymorphisms (SNPs) of the myostatin gene segregating in these lines were identified by designing specific primers, amplifying individual DNA in each line by polymerase chain reaction, cloning, sequencing and aligning the corresponding products. Individual sires were genotyped for five identified SNPs which contributed to eight haplotypes. Frequencies of SNP alleles and haplotypes differed between lines. Using the allele substitution effect model, the myostatin SNPs were found to have significant (P < 0.031) associations with growth, mortality, blood oxygen and hen antibody titer to infectious bursal disease virus vaccine, although the associations were not often consistent across lines. These results suggest that the myostatin gene has pleiotropic effects on broiler performance.

Simultaneous genotyping to detect myostatin gene polymorphism in beef cattle breeds

SummaryThe myostatin gene codes for a growth factor involved in muscle development, and polymorphism in this gene can have important economic consequences. Nine mutations affecting the amino‐acidic sequence have already been described, six of which are disruptive, inactivating the protein and causing bovine muscular hypertrophy. As the number of known mutations grows, it is necessary to develop a simple, routinely usable technique able to screen individuals in all populations. The oligonucleotide ligation assay (OLA) is proposed here for the rapid genotyping of the nine mutations known affecting the coding sequence in the main breeds of beef cattle. This technique showed its ability to reveal the genotype of individuals being a good tool to determine the frequency of each mutation in a population. The procedure is very flexible as new mutations can be added and removed at any time. Depending on the genotype of each individual, the technique allows breeders to make quick decisions on matings and general selection tendencies.