Evaluation of the porcine melanocortin 4 receptor (MC4R) gene as a positional candidate for a fatness QTL in a cross between Landrace and Hampshire

Melanocortin 4 receptor (MC4R) is expressed in the appetite-regulating areas of the brain where it is central in the regulation of feed intake and energy balance. A mutation in MC4R causing an Asp298Asn substitution has been associated with fatness, high daily gain and feed intake in the pig. In a previously performed genome scan based on a Hampshire x Landrace cross, we detected one quantitative trait loci (QTL) affecting carcass fat/meat ratio and one QTL affecting the biceps femoris muscle, both close to the position of MC4R on porcine chromosome 1. In this study, the two lines were found to be close to fixation for alternative alleles of the Asp298Asn polymorphism. Additional QTL analyses supported our hypothesis of MC4R as a positional candidate gene but only for the fat/meat QTL. The Asp298Asn polymorphism was also evaluated as a selection target for daily gain in a Danish pig breeding population that included four breeds (Hampshire, Duroc, Landrace and Yorkshire). Over a 12-year period (1990-2002), a significant increase in the allele frequency of 298Asn was found in Landrace and Duroc, whereas a non-significant decrease in the 298Asn allele frequency was observed in Yorkshire. The Hampshire breed was fixed for the 298Asn allele in 1990. The high 298Asn allele frequencies in Hampshire, Landrace and Duroc are most likely due to selection for daily gain, whereas selection for daily gain in the Yorkshire breed apparently focuses on other loci.     

Melanocortin-4 receptor (MC4R) genotypes have no major effect on fatness in a Large White x Wild Boar intercross

The melanocortin-4 receptor (MC4R), a G-protein coupled receptor, is implicated in mediating the effect of leptin on food intake and energy balance. A previous candidate gene study reported an association between an MC4R missense mutation (Asp298Asn) and fatness, growth and feed intake in pigs. To assess this association further, we analysed the segregation of this missense mutation in relation to variation in fatness traits using a Wild Boar x Large White intercross. The Wild Boar and Large White founders were homozygous for different MC4R alleles. The MC4R was assigned to the expected region on pig chromosome 1. The statistical evaluation did not reveal any indication of a significant effect on fatness related traits in this pedigree.     

An effect of a missense mutation in the porcine melanocortin-4 receptor (MC4R) gene on production traits in Polish pig breeds is doubtful

A missense G-A SNP in the porcine melanocortin-4 receptor (MC4R) gene that causes an Asp-Asn substitution at position 298 of the corresponding MC4R protein is considered to be economically important, although published results on its effect are inconsistent. We analysed the association of this MC4R polymorphism with production traits in 679 gilts from two breeds, Polish Large White (PLW) and Polish Landrace (PL), as well as one synthetic line 990. The frequency of the A allele differed significantly among the breeds with frequencies of 0.76, 0.29 and 0.16 in PLW, PL and line 990 respectively. There was no evidence of an effect of this polymorphism on daily food intake, backfat thickness or abdominal fat. The A allele was correlated with higher test daily gains and lower levels of intramuscular fat in PL, and increased levels of intramuscular fat in PLW.     

The Asp298Asn missense mutation in the porcine melanocortin-4 receptor ( MC4R) gene can be used to affect growth and carcass traits without an effect on meat quality

A promising tool to improve daily gain in pigs is the missense mutation (Asp298Asn) in the melanocortin-4 receptor (MC4R) gene, especially in the Belgian pig industry where the slow-growing Piétrain breed is very frequently used as the sire breed. The MC4R is expressed in the appetite-regulating region of the brain where it regulates feed intake and energy balance. The mutation has been associated with differences in fatness, daily gain and feed intake. However, less information on the correlated effects on meat quality is available. In order to evaluate the influence of the MC4R mutation on carcass and meat quality parameters, a total of 1155 pigs of a four-way cross were slaughtered at an average live weight of 109 kg, and data about daily live-weight gain, carcass and meat quality were collected. Allelic frequencies were 0.69 for the G-allele (298Asp variant or well-conserved variant) and 0.31 for the A-allele (298Asn variant or the mutated variant). Barrows and gilts were almost equally distributed in this population with, respectively, 49.9% and 50.1%. Moreover, independent of this mutation, the relationship between average daily gain (ADG) and carcass on the one hand and meat quality traits on the other hand was evaluated in this population. A significant positive influence of the MC4R mutation on ADG (P < 0.001) was found, accompanied by a higher fat thickness (P < 0.05) and a lower carcass lean meat content (P < 0.01), whereas muscle thickness and carcass conformation traits were not affected. The effects on meat quality traits were not significant, except for a lower shear force (P = 0.054) and a higher intramuscular fat content (P = 0.052) in AA animals. In the longissimus, pH and pork quality meter (PQM) values were not influenced, and effects on drip loss and colour were not apparent. Residual correlation coefficients between ADG and carcass lean meat content on the one hand and meat quality traits on the other hand were generally very low (|r|>0.1). Higher ADG, higher carcass fat thickness and lower carcass lean meat content were correlated with slightly lower shear force values (|r|～0.1, P < 0.05). In conclusion, in the studied population, the Asp298Asn mutation in the MC4R gene was associated with improved daily gain, higher carcass fatness and almost no effect on meat quality traits.     

Association of the melanocortin 4 receptor with feed intake and daily gain in F2 Mangalitsa x Piétrain pigs

The melanocortin 4 receptor (MC4R) is a key factor in the regulation of energy balance and body weight. Hence it is a candidate for feed intake and energy homeostasis-related traits. Studies in humans and swine have revealed several sequence variants in the gene that are associated with some of these traits. In pigs the coding non-synonymous missense variant Asp298Asn in MC4R has been associated with feed intake, fatness and growth. Here we confirm the association of this Piétrain-derived polymorphism with feed intake and daily gain in the F2 generation of a Mangalitsa x Piétrain cross. In one Piétrain founder animal, we detected an additional non-synonymous missense variant Arg236His. Thus, the MC4R gene could be a useful marker for increased growth in the relatively slow-growing Piétrain breed.     

Detailed characterization of the porcine MC4R gene in relation to fatness and growth

In contrast to the human MC4R gene, where multiple variants have been described, several of which are associated with appetite and obesity, few MC4R variants have been reported in the pig. The most interesting polymorphism reported to date in the pig is p.Asp298Asn, which is significantly associated with variation in growth and fatness traits in most breeds and crosses. However, some reports have seemingly failed to confirm this association. The discrepancy of p.Asp298Asn associations in some pig populations suggested that further discovery of SNPs in MC4R would be useful. Utilizing the recently released pig genome sequence information, we obtained the whole MC4R genome sequence and detected five additional SNPs, a variable (CA) _n repeat and a C indel in the ISU Berkshire × Yorkshire pig resource family. Linkage disequilibrium (LD) analysis revealed that the additional five SNPs were not in strong LD with p.Asp298Asn, but single marker association analysis indicated that they were significantly ( P  <   0.05) associated with fatness measures and very highly significantly ( P  <   0.0001) associated with average daily gain on test (ADGTEST). Three major haplotypes were identified and the subsequent association analyses suggested that the two non-synonymous SNPs had different effects, e.g. p.Arg236His influenced back fat and growth on test while p.Asp298Asn was primarily associated with variation in growth rate in this population. An interaction effect between these two SNPs was found for ADGTEST, which may partly explain some of the previous discrepancies reported for MC4R in different pig populations . Examination of the p.Arg236His polymorphism in populations where the effect of p.Asp298Asn is limited is warranted.     

Correlation between the Asp298Asn polymorphism of mc4r gene and back fat width in pigs of large white bred

Asp298Asn correlation between the mc4r gene polymorphism and development of such trait as back fat width in Large White pigs belonging to AF "Orzhitska" population was studied. The tendency of forming thinner back fat layer in animals possessing AG genotype was detected. Any reliable differences in back fat width between animal groups with various mc4r genotypes caused by the Asp298Asn polymorphism were not detected. A possible reason for the absence of statistically reliable difference may be heterogeneous origin of the studied animals. Using Asp298Asn polymorphism of mc4r gene in selecting the Large White AF "Orzhitska" pig population is unreasonable.     

The Asp298Asn polymorphism of melanocortin-4 receptor (MC4R) in pigs: evidence for its potential effects on MC4R constitutive activity and cell surface expression

In humans and mice, melanocortin receptor 4 (MC4R) and melanocortin receptor accessory protein 2 (MRAP2) can form a complex and control energy balance, thus regulating body weight and obesity. In pigs, a missense variant (p.Asp298Asn) of MC4R has been suggested to be associated with growth and fatness; however, the effect of Asp298Asn substitution on MC4R function is controversial, limiting its application in animal breeding. Here we examined the effect of this polymorphism on MC4R constitutive activity, cell surface expression and signaling, and its interaction with MRAP2 in pigs. We found that: (i) both pig MC4R^Asp and MC4R^Asn can be activated by its ligands (α-MSH and ACTH) and stimulate cAMP/PKA signaling pathway, as detected by pGL3–CRE–luciferase reporter assay, indicating that, like pMC4R^Asp, pMC4R^Asn is coupled to the cAMP/PKA signaling pathway; (ii) compared with pMC4R^Asp, pMC4R^Asn loses the basal constitutive activity and shows a decreased surface expression, as detected by dual-luciferase reporter assay and Nano-HiBiT system; (iii) as in other vertebrates, both pMC4R^Asp and pMC4R^Asn can interact with pMRAP2, thus decreasing receptor surface expression and enhancing ligand sensitivity, although, in contrast to pMC4R^Asp, the basal constitutive activity of pMC4R^Asn cannot be affected by pMRAP2; and (iv) RNA-seq data analysis revealed a co-expression of MC4R and MRAP2 in pig hypothalamus. Taken together, our data provide convincing evidence that Asp298Asn substitution decreases the constitutive activity and cell surface expression of MC4R or MC4R–MRAP2 complex, which may affect energy balance and be a valuable selection marker for breeding programs in pigs.     

Association of the Asp298Asn polymorphism in the mc4r gene with back fat thickness in pigs of the large white breed

The association of the Asp298Asn polymorphism in the mc4r gene with the development of traites such as spinal fat thickness in pigs of the Large White breed from AF “Orzhitska” population was investigated. A tendency towards the development of thinner spinal fat was found in animals with the AG genotype. No significant difference was detected in the spinal fat thickness between animal groups with different genotypes of the mc4r gene caused by Asp298Asn polymorphism. The heterogeneous origin of the studied animals can be a possible cause of the absence of any statistically significant difference. A conclusion was reached that it was not expedient to use Asp298Asn polymorphism of the mc4r gene in the selection of pigs of the Large White breed from AF “Orzhitska” population.     

Association od the DdeI growth hormone gene polymorphism with some performance traits in Polish Large White and Czech Large White x Polish Large White pigs

The genotypes of growth hormone gene polymorphisms (GH-DdeI, GH-MspI, GH-HaeII, GH-ApaI, GH-CfoI) were determined in 78 pigs [Czech Large White sires (CLWsire line) x Polish Large White (PLW) sows, Polish Large White sires x Polish Large White sows], by the PCR-RFLP method. Preliminary studies found only GH DdeI polymorphism to be associated with performance traits. The associations of this polymorphism with growth and carcass traits were investigated. The linear model included the effects of candidate genes, genetic groups, sex and linear covariables of age at slaughter and body weight at weaning. The DdeI polymorphism of the GH gene showed associations with carcass length (P 相似文献   

Peri-partum posture and behaviour of gilts and the location of their piglets in lines selected for components of efficient lean growth

Peri-partum posture and behaviour of gilts from lines selected for different components of efficient lean growth were studied to determine if behavioural changes may have been associated with the observed responses in reproductive performance. The proportions of time that gilts expressed defined posture and behaviour traits and the locations of their piglets were determined from video recordings of observations made at 5min intervals in the period extending from 2h pre-farrowing to 2h post-farrowing. The 137 gilts studied were from four pairs of Large White lines which had been divergently (high and low) selected for either daily food intake (DFI), lean food conversion efficiency (LFC), lean growth rate on ad libitum feeding (LGA) or lean growth rate on a restricted feeding scale (LGS).Almost all the significant (P<05) changes occurred in the LGS pair of lines. In the pre-farrowing period, relative to the low LGS gilts, high LGS gilts spent a higher proportion of their time lying on their sides (0.92 versus 0.69), and less time in the upright postures of standing, sitting or lying on their bellies (0.08 versus 0.33) and engaging in nesting behaviour (0.02 versus 0.10). During farrowing, high LGS gilts again lay on their sides more often than low LGS gilts (0.96 versus 0.80) and were upright less often (0.04 versus 0.20). High LGS gilts changed posture less often than low LGS gilts (0.05 versus 0.31) but were more often alert (0.79 versus 0.61). During farrowing, high LGS piglets were seen less often at their mother's head, back and vulva or at the creep than low LGS piglets (0.06 versus 0.15). Post-farrowing, there were no significant differences between the lines, almost all gilts lying on their sides with their piglets at the udder. Divergent selection for components of efficient lean growth rate on ad libitum feeding was not associated with consistent responses in gilt posture and behaviour or in piglet location. Selection for high lean growth on restricted feeding had effects on gilt posture and behaviour which may have been beneficial to her welfare and that of her piglets.     

Association of melanocortin 4 receptor (MC4R) and high mobility group AT-hook 1 (HMGA1) polymorphisms with pig growth and fat deposition traits

The aim of this study was to analyse the combined effect of melanocortin 4 receptor (MC4R) and high mobility group AT-hook 1 (HMGA1) polymorphisms on growth and fatness traits in Duroc pigs. No significant interaction was observed between MC4R and HMGA1 for back-fat traits. An additive mode of inheritance of both gene effects was found for average daily gain and lean meat content. Maximum mean differences from combined genotypic effects were over 2 mm for back fat, 70 g/day for average daily gain and 2% for lean meat content. Therefore, utilization of polymorphisms in both MC4R and HMGA1 for marker-assisted selection could result in an economic benefit to the pig industry.     

A missense variant of the porcine melanocortin-4 receptor (MC4R) gene is associated with fatness, growth, and feed intake traits

Our knowledge of the genetic factors affecting obesity is increasing, but information about the individual gene effects remains limited in humans as well as in animal models. The melanocortin-4 receptor gene (MC4R) has been implicated in the regulation of feeding behavior and body weight in humans and mice. We have studied MC4R as a candidate gene for the control of economically important growth and performance traits in the pig. A missense mutation was identified in a region highly conserved among melanocortin receptor (MCR) genes. To determine whether there was an association of this MC4R polymorphism with phenotypic variation, we tested the mutation in a large number of individual animals from several different pig lines. Analyses of growth and performance test records showed significant associations of MC4R genotypes with backfat and growth rate in a number of lines as well as feed intake overall. It is probable that the variant amino acid residue of the MC4R mutation (or a closely linked mutation) causes a significant change of the MC4R function. These results support the functional significance of a pig MC4R missense mutation and suggest that comparative genomics based on model species may be equally important for application to farm animals as they are for human medicine. Received: 21 June 1999 / Accepted: 9 September 1999     

Analysis of Association Between the MUC4 g.8227C>G Polymorphism and Production Traits in Italian Heavy Pigs Using a Selective Genotyping Approach

In pigs, susceptibility to enterotoxigenic Escherichia coli (ETEC) K88 strains (locus F4bcR) is determined by a dominant allele, with the recessive allele determining resistance. The susceptible allele also appeared to be associated with higher growth rate even with discordant results. A single nucleotide polymorphism (SNP) in exon 7 of the mucin 4 (MUC4) gene (DQ848681:g.8227C>G), shown to be in close linkage disequilibrium with the F4bcR locus, has been used as marker to identify susceptible pigs, substituting invasive villous adhesion tests. We herein analyzed this SNP in Italian local breeds and applied a selective genotyping approach in Italian Large White, Italian Landrace, and Italian Duroc comparing allele frequency distribution in groups of pigs with extreme estimated breeding values (EBV) for average daily gain (ADG) and backfat thickness (BFT) to evaluate if this marker is associated with these traits. Allele G (associated with susceptibility to ETEC) was associated with higher ADG and BFT in Italian Large White (P?=?6.66E-04 and P?=?0.012, respectively) and higher ADG in Italian Landrace (P?=?7.23E-12). This polymorphism was poorly informative in Italian Duroc. Antagonistic associations of the MUC4?g.8227C>G alleles on susceptibility to ETEC and growth performances evidence the complexity of applying marker assisted selection in pig breeding.     

Polymorphisms in MC4R gene and correlations with economic traits in cattle

MC4R contributes to the control of food intake and energy expenditure, and single nucleotide polymorphisms (SNPs) in the MC4R gene have clearly been associated with backfat depth, feed intake and growth rate in pig. Our objectives were to scan the complete coding region by sequencing in samples from eight cattle breeds, to estimate the frequency of the SNPs in the MC4R gene and to determine if individual genotypes were associated with several economic traits. Five polymorphisms were detected at position 19 (C/A), 20 (A/T), 83(T/C), 128 (G/A), and 1069 (G/C), and the last one was significantly associated with backfat thickness value (P < 0.01, n = 245). The linkage disequilibrium analysis indicated that the SNP markers C19A, A20T, T83C and G128A were completely linked (r ² = 1).     

The cholecystokinin type A receptor g.179A>G polymorphism affects feeding rate

A polymorphism within the 5' untranslated region of the cholecystokinin type A receptor ( CCKAR ) gene has been shown to affect feed intake and growth in commercial pig lines. To further investigate the phenotype of animals carrying alternative alleles at this polymorphism, we genotyped animals from a distinct segregating commercial line and an experimental cross F₂ population, both with electronically recorded feeding pattern data. The data indicate that the daily feed intake increasing effect of the DQ496228:g.179G allele is mediated through a faster rate of feed intake, without evidence for an effect on other feeding behaviour traits.     

Analysis of Association Between the MUC4 g.8227C>G Polymorphism and Production Traits in Italian Heavy Pigs Using a Selective Genotyping Approach

In pigs, susceptibility to enterotoxigenic Escherichia coli (ETEC) K88 strains (locus F4bcR) is determined by a dominant allele, with the recessive allele determining resistance. The susceptible allele also appeared to be associated with higher growth rate even with discordant results. A single nucleotide polymorphism (SNP) in exon 7 of the mucin 4 (MUC4) gene (DQ848681:g.8227C>G), shown to be in close linkage disequilibrium with the F4bcR locus, has been used as marker to identify susceptible pigs, substituting invasive villous adhesion tests. We herein analyzed this SNP in Italian local breeds and applied a selective genotyping approach in Italian Large White, Italian Landrace, and Italian Duroc comparing allele frequency distribution in groups of pigs with extreme estimated breeding values (EBV) for average daily gain (ADG) and backfat thickness (BFT) to evaluate if this marker is associated with these traits. Allele G (associated with susceptibility to ETEC) was associated with higher ADG and BFT in Italian Large White (P = 6.66E-04 and P = 0.012, respectively) and higher ADG in Italian Landrace (P = 7.23E-12). This polymorphism was poorly informative in Italian Duroc. Antagonistic associations of the MUC4 g.8227C>G alleles on susceptibility to ETEC and growth performances evidence the complexity of applying marker assisted selection in pig breeding.     

A single nucleotide polymorphism in the porcine cathepsin K (<Emphasis Type="Italic">CTSK</Emphasis>) gene is associated with back fat thickness and production traits in Italian Duroc pigs

Cathepsin K (CTSK) was selected as a candidate gene for fat deposition in pigs because recently, in human and mouse, it was shown that this lysosomal proteinase is an obesity marker. A single nucleotide polymorphism (SNP) was identified in intron 4 of the porcine CTSK gene (g.15G>A; FM209043). Allele frequencies of this polymorphism were analysed in seven pig breeds. Radiation hybrid mapping confirmed the localization of CTSK to porcine chromosome 4, close to the FAT1 QTL region. Three populations of pigs (one Italian Large White and two Italian Duroc groups of pigs) were selected for association analysis. In the Italian Large White breed the g.15G>A SNP was not informative. Association analysis including all Italian Duroc pigs showed that the CTSK marker was associated with back fat thickness and lean cuts (P < 0.01), and average daily gain and feed:gain ratio (P < 0.05) estimated breeding values.     

Genetic parameters and responses of performance and body composition traits in pigs selected for high and low growth rate on a fixed ration over a set time

Two lines of Large White pigs of common genetic origin were divergently selected over four years for high and low growth rate during a 6 week post-weaning test period in which all pigs were fed the same total amount of food (80% of estimated ad libitum intake). Genetic parameters and direct and correlated responses in performance and carcass traits were estimated on 2884 pigs with pedigrees comprising a total of 5324 animals, with restricted maximum likelihood and best linear unbiased prediction methods applied to a multi-trait animal model. Estimates of heritability (± SE) were 0.19 ± 0.04 for lifetime daily gain (LDG), 0.16 ± 0.03 for test daily gain (TDG), 0.25 ± 0.04 for ultrasound P2 backfat (UBF) and 0.16 ± 0.03 for food conversion ratio during test (TFC), and 0.15 ± 0.04 for daily carcass weight gain (CDG), 0.43 ± 0.06 for carcass backfat (CFT) and 0.40 ± 0.06 for carcass lean percentage (LEAN). Common litter effects for TDG, UBF and TFC were less than 5% and for LDG, 17% of total phenotypic variance. Genetic correlations between performance and carcass traits were moderately to highly favourable. After four years of divergent selection for growth rate, the selection responses in estimated breeding value (EBV) for TDG were 40.14 and -41.11 g (SED 2.95) for the high and low growth lines, respectively. The regressions of EBV on year of birth, indicate that the annual genetic trend for TDG, was 8.73 g/yr in the high and -8.48 g/yr in the low lines (P < 0.001). Correlated genetic responses in the high and low lines respectively were 5.28 g and -12.40 g (SED 2.09) in LDG, -0.35 mm and 0.56 mm (SED 0.009) in UBF, -0.145 units and 0.185 units (SED 0.012) in TFC, 3.17 g and -10.97 g (SED 1.53) in CDG, -1.13 mm and 1.01 mm (SED 0.155) in CFT and 1.24% and -1.27% (SED 0.150) in LEAN. It was concluded that selection for increased post-weaning daily gain on a ration of fixed amount reduces the age at slaughter and the level of backfat and increases the efficiency of food utilisation, weight and leanness of pig carcasses.