Short communication: Locus-specific interrelations between gene expression and DNA methylation patterns in bovine mammary gland infected by coagulase-positive and coagulase-negative staphylococci.

Pathogens are able to alter the cell cycle program and immune response of the host by changing the transcription and epigenetics of genes responsible for cell cycle control and inflammation. In this regard, we evaluated interrelations between DNA methylation and expression of autophagy, apoptosis, and lipid metabolism-related genes in a sample set of mammary gland secretory tissue sections derived from bovine mammary glands infected with coagulase-negative and coagulase-positive staphylococci. We assessed relative transcript abundance and DNA bisulfite sequencing in loci of the ATG5, IGF1R, TERT, and DGAT1 genes. Lack of DNA methylation in ATG5 and DGAT1 loci might be associated with maintenance of ATG5 and DGAT1 expression regardless of the health status of bovine mammary gland. Complete methylation of intragenic CpG regions in the IGF1R locus was apparently not related to the presence of its transcript in the investigated udder parenchyma samples. Detected hypermethylation of the TERT upstream element was associated with a small amount of TERT mRNA in bovine mammary gland, regardless of the presence, or absence, of the pathogen. A significant decrease in TERT gene expression in tissue sections of mammary gland free of bacteria and in those infected with coagulase-positive staphylococci was observed in parenchyma samples infected with coagulase-negative staphylococci. Two possible explanations are the direct involvement of the TERT gene in the etiology of bovine mastitis or the increase of TERT mRNA due to activation of the MAPK signaling pathway in response to release of exotoxins by coagulase-negative bacteria in the bovine mammary gland.

Structural and functional analysis of the signaling lymphocytic activation molecule family 7 (SLAMF7) gene in response to infection with coagulase-negative and coagulase-positive staphylococci.

Splice variants of the signaling lymphocytic activation molecule family 7 (SLAMF7) gene have been identified, and differences in the expression of this gene have been demonstrated at the mRNA level in the mammary glands of healthy and mastitis-infected dairy cows. At the same time, significant associations have been found between a deletion in the SLAM7 gene exon, the occurrence of different splice variants, and the occurrence of mastitis in one group of dairy cows. An expression study was conducted on 40 Polish Holstein-Friesian dairy cows of the Black and White variety (group I). Milk samples were taken for microbiological analysis 2 d before slaughter and examined for the presence of bacteria. Immediately after slaughter, mammary tissue samples were taken and divided into 3 groups according to the health status of the mammary gland: healthy (without pathogenic bacteria in milk), coagulase-negative staphylococci (CNS), and coagulase-positive staphylococci (CPS). Based on different SLAMF7 gene DNA fragments, 2 alternative variants of this gene (V1 and V2) and complete gene expression were identified. Separate analyses performed for each isoform showed that the health status of the cow was strongly associated with the expression level of individual variants. The highest expression was detected for the SLAMF7 complete amplicon in healthy cows, and in the CNS and CPS cows the expression of this variant was also higher than V1 and V2. Sanger sequencing was applied to detect the polymorphism/indel variant in the second exon of the SLAMF7 gene probably having the greatest effect on the protein structure and function of SLAMF7. Two genotypes were detected: AA (wild-type) and AB (insertion A). In healthy cows, the frequency of homozygotes AA was higher than the heterozygotes, whereas in the infected animals, the genotypic distribution was the opposite. An association analysis between the identified polymorphism and production traits-including somatic cell count, as well as lactose, protein, and casein content and yield as indicators of subclinical mastitis occurrence-was performed on the group II cows (166 Polish Holstein-Friesian dairy cows). Unfortunately, due to the low number of AB animals, no relationship was demonstrated between genotype in the second exon and the health status of cows. Additionally, the difference in the percentage of SLAMF7-targeted DNA methylation between the groups of animals was not significant, with an average of ∼66 to 68%.

Association between single nucleotide polymorphisms in GPAT1 locus and pork quality in pigs.

GPAT1 gene is considered to be a genetic marker for intramuscular fat content. The GPAT enzymes catalyze the first step in triacylglycerol synthesis. In the present study, the search for polymorphisms within the pig GPAT1 gene locus as well as association analyses between SNPs and meat quality traits were performed. The association analysis demonstrated that g.133513422C > T polymorphism affected IMF content in LL, SEMI post-mortem pH and shear force of cooked LL (p ≤ .05). While the association of g.133476803 T > C polymorphism was shown concerning IMF content, meat color (L*), shear force and energy of raw meat as well as some meat texture parameters (hardness, springiness, chewiness) (p ≤ .05). The g.133476733C > T SNP was significantly associated with LL 24 h post-mortem pH, raw meat toughness and TPA hardness and chewiness (p ≤ .05). The obtained results are promising but to confirm if the GPAT1 gene can be considered to be a genetic marker for improving the quality of pork, further research is required.

The Genetics of Racing Performance in Arabian Horses.

Arabian horses are commonly believed to be one of the oldest and most influential horse breeds in the world. The high financial benefits obtained from races tend to search for genetic markers strongly correlated with the results achieved. To date, the modern approaches such as transcriptome, miRNAome, and metabolome analyses have been used to investigate the genetic background of racing performance as well as endurance capacity in Arabians. The analysis of polymorphisms at the genome level has also been applied to the detection of genetic variants associated with exercise phenotype in the Arabian breed. The presented review summarizes these findings, with a focus on the genetics underlying flat racing and endurance performance traits in different Arabian horse populations.

Sequence analysis and expression profiling of the equine ACTN3 gene during exercise in Arabian horses.

The ACTN3 gene codes for α-actinin-3, a protein localized in the Z-line in the skeletal muscle. Actinin-3 is critical in anchoring the myofibrillar actin filaments and plays a key role in muscle contraction. ACTN3 (α-actinin-3) cross-links glycogen phosphorylase (GP), which is the key enzyme catalysing glycogen metabolism. The aim of present study was to establish the expression level of the ACTN3 gene (for both isoforms separately and together in the gene expression analysis) in the gluteus medius muscle in order to verify if the α-actinin-3 gene can be related to training intensity in Arabian horses. A structural analysis of the ACTN3 gene was performed simultaneously to identify polymorphisms that can be related to racing performance traits. Our results showed the significant decrease (p < 0.05) of ACTN3 expression in the skeletal muscle of Arabian horses during the training periods preparing for flat-racing, and this decrease differed by the intensity of the exercises. The highest mRNA abundance measured for all ACTN3 genes was detected in the muscle of untrained horses, while the lowest expression was identified at the end of the racing season when horses had fully adapted to the physical effort. This gene expression profile was confirmed for both ACTN3 isoforms. The analysis of the ACTN3 sequence allowed us to identify 14 polymorphisms, which were localized in the promoter region, the 5'UTR (7 SNPs), exons (2 SNPs) and introns (5 SNPs). Two of them, a novel c.2334C>T - splice variant and the g.1104G>A polymorphism in the promoter region, were proposed as the causative mutations that might affect gene expression. The presented gene expression analyses indicated the significant role of the ACTN3 gene in adaptation to physiological effort in horses. Due to previous reports and our findings, further studies should be conducted to verify the usage of the ACTN3 gene as a potential genetic marker for determining exercise performance in Arabian horses and other horse breeds.

Molecular characterization of the apoptosis-related SH3RF1 and SH3RF2 genes and their association with exercise performance in Arabian horses.

BACKGROUND: Apoptosis plays an important role in the regulation of healthy tissue growth and development as well as in controlling the maintenance of homeostasis in exercising muscles. During an intensive physical effort, the regulation of cell death by apoptosis results in the replacement of unaccustomed muscle cells by new cells that are better suited to exercise. The aim of this study was to determine the expression of two genes (SH3FR1 and SH3RF2) that control apoptosis in muscle tissues during training periods characterized by different intensities. The gene expression levels were estimated using real-time PCR method in skeletal muscle biopsies collected from 15 Arabian horses (untrained, after an intense gallop phase, and at the end of the racing season). An association study was performed on 250 Arabian horses to assess the effect of the SH3RF2:c.796 T > C (p.Ser266Pro) variant on race performance traits in flat gallop-racing. RESULTS: A gene expression analysis confirmed a significant decrease (p < 0.01) in the anti-apoptotic SH3RF2 (POSHER) gene during training periods that differed in intensity. The highest SH3RF2 expression level was detected in the muscles of untrained horses, whereas the lowest expression was identified at the end of the racing season in horses that were fully adapted to the exercise. A non-significant decrease in SH3RF1 gene expression following the training periods was observed. Moreover, a serine substitution by proline at amino acid position 266 (CC genotype) was negatively associated with the probability of winning races, the number of races in which a horse occurred and the financial value of the prizes. Horses with the TT genotype achieved the highest financial benefits, both for total winnings and for winnings per race in which the horses participated. CONCLUSIONS: The present study showed the supposed regulation mechanism of exercise-induced apoptosis in horses at the molecular level. The identified SH3RF2: c.796 T > C missense variant was associated with selected racing performance traits, which is important information during the evaluation of horses' exercise predisposition. The association results and frequencies of the CT and TT genotypes suggest the possibility of using SH3RF2 variant in selection to improve the racing performance of Arabian horses.

Transcriptomic hallmarks of bone remodelling revealed by RNA-Seq profiling in blood of Arabian horses during racing training regime.

The impact of exercises on young developing organisms is still of interest to researchers. Similarly like Thoroughbreds, Arabian horses competing at the race track. The high percent of lameness and loss of days in training are often the result of weakness in the condition of the musculoskeletal system. The objective of the presented study was to identify by RNA-Seq method, the possible skeletal system originating transcriptomic profile in peripheral blood of Arabian horses undergoing race training. Obtained results showed that one of the most significantly deregulated pathway involved in bone homeostasis was those involved in osteoclast differentiation. Among the significantly expressed molecules, we recognized twelve genes potentially involved in the metabolism of the skeletal system: BGLAP, CTSK, TYROBP, PDLIM7, SLC9B2, TWSG1, NOTCH2, IL6ST, VAV3, NFATc1, CLEC5A, TXLNG. The panel of identified genes should be evaluated as candidate biomarkers for bone homeostasis indicators of Arabians performing on race tracks to assess bone remodelling states during training for race track competitions.

Detection of genetic variants between different Polish Landrace and Pulawska pigs by means of RNA-seq analysis.

Variant calling analysis based on RNA sequencing data provides information about gene variants. RNA-seq is cheaper and faster than is DNA sequencing. However, it requires individual hard filters during data processing due to post-transcriptional modifications such as splicing and RNA editing. In the present study, RNA-seq transcriptome data on two Polish pig breeds (Pulawska, PUL, n = 8, and Polish Landrace, PL, n = 8) were included. The pig breeds are significantly different with regard to meat qualities such as texture, water exudation, growth traits and fat content in carcasses. A total of 2451 significant mutations were identified by a chi square tests, and functional analysis was carried out using Panther, KEGG and Kobas. Interesting missense gene variants and mutations located in regulatory regions were found in a few genes related to fatty acid metabolism and lipid storage such as ACSL5, ALDH3A2, FADS1, SCD, PLA2G12A and ATGL. A validation of mutational influences on pig traits was performed for ALDH3A2, ATGL, PLA2G12A and MYOM1 variants using association analysis including 215 pigs of the PL and PUL breeds. The ALDH3A2ENSSSCT00000019636.2:c.470T>C polymorphism was found to affect the weight of the ham and loin eye area. In turn, an ENSSSCT00000004091.2:c.2836G>A MYOM1 mutation, which could be implicated in myofibrillar network organisation, had an effect on meatiness and loin texture parameters. The study aimed to estimate the usefulness of RNA-seq results for a purpose other than differentially expressed gene analysis. The analysis performed indicated interesting gene variants that could be used in the future as markers during selection.

Transcriptome analysis of equine sarcoids.

Equine sarcoids are the most commonly detected skin tumours in Equidae. In the present research, a comparative transcriptomic analysis was performed which aimed at looking inside a tumour biology and identification of the expression profile as a potential source of cancer specific genes useful as biomarkers. We have used Horse Gene Expression Microarray data from matched equine sarcoids and tumour-distant skin samples. In total, 901 significantly differentially expressed genes (DEGs) between lesional and healthy skin samples have been identified (fold change ≥ 2; P < 0.05). The large subset of DEGs, with decreased expression, was associated with a suppression of malignant transformation, whereas several overexpressed genes were involved in the processes associated with growth and progression of a tumour or immune system activity. Our results, as a first to date, showed comprehensive transcriptome analysis of skin tumour in horses and pinpointed significant pathways and genes related with oncogenesis processes.

Identification of genome-wide selection signatures in the Limousin beef cattle breed.

The study is aimed at identifying selection footprints within the genome of Limousin cattle. With the use of Extended Haplotype Homozygosity test, supplemented with correction for variation in recombination rates across the genome, we created map of selection footprints and detected 173 significant (p < 0.01) core haplotypes being potentially under positive selection. Within these regions, a number of candidate genes associated inter alia with skeletal muscle growth (GDF15, BMP7, BMP4 and TGFB3) or postmortem proteolysis and meat maturation (CAPN1 and CAPN5) were annotated. Noticeable clusters of selection footprints were detected on chromosomes 1, 4, 8 and 14, which are known to carry several quantitative trait loci for growth traits and meat quality. The study provides information about the genes and metabolic pathways potentially modified under the influence of directional selection, aimed at improving beef production characteristics in Limousin cattle.

Genome-wide RNA-Seq analysis of breast muscles of two broiler chicken groups differing in shear force.

In this study, a whole transcriptome analysis of breast muscles was conducted in broiler chicken groups differing in shear force. Shear force is a determinant of tenderness, which in turn is one of the most important parameters of meat quality in chickens. In our analysis, a total of 11,560 transcripts and 9824 genes per sample were identified. In chickens with more tender meat, up-regulation of 19 genes and down-regulation of 49 genes was observed. The up-regulated gene group included the ASB2 gene, which is probably involved in the meat conversion process, as its product results in the degradation of filamins, proteins which form muscle fibres. In the down-regulated gene group, genes which play a role in lipogenesis (THRSP, PLIN1) and in collagen synthesis (P4HA3, LEPREL4, PCOLCE2, COL16A1, COL20A1, VWA1) were detected. Their presence suggests the involvement of the extracellular matrix in the determination of meat tenderness. Thus, our study identified a pool of genes that may participate in the tenderisation process in broiler chickens.

Association of calpastatin gene polymorphisms and meat quality traits in pig.

Calpastatin is associated with the rate of post mortem degradation of structural proteins due to the regulation of calpain activity. In the present research, the associations between polymorphisms within 6th intron of porcine CAST gene and several meat quality traits were analyzed. The CAST gene polymorphisms affected meat colour, pH, water holding-capacity (WHC) and texture parameters (toughness, firmness, cohesiveness, chewiness, and resilience) measured in longissimus dorsi and semimembranosus muscles. The analysis performed on the most numerous breeds maintained in Poland, suggested that the most interesting polymorphisms were CAST/HpaII and CAST/RsaI, which had the greatest effect on WHC regardless of the breed analyzed and had an effect on meat pH, firmness and toughness for most breeds. Interestingly, for almost all breeds, the significant effect of both mutations on intramuscular fat content (IMF) was detected. The provided data confirmed the use of CAST gene as a genetic marker in breeding programmes which allows performing a selection focussed on improving the quality of pork.

Association between LEPR and MC4R genes polymorphisms and composition of milk from sows of dam line.

The polymorphisms of LEPR and MC4R genes are involved in appetite control mechanisms and indirectly associated with level of fat content in pig carcasses. Therefore, the aim of our study was to determine if both polymorphisms have an effect on components of colostrum and milk of sows. In our study we used gilts of two Polish breeds: Polish Landrace and Polish Large White, which belong to dam-line in Polish breeding. Colostrum and milk of sows were collected in 7, 14 and 21 day of lactation to assay solids, total protein, fat and lactose. The obtained results showed, that the observed mutation (G/A 1426 MC4R) had a significant effect mainly on the fat and solids content of colostrum. Animals with the MC4R (AA) genotype had 2.13 and 1.91 % (P ≤ 0.01) lower fat content of colostrum compared to sows with the MC4R (GG) genotype and heterozygous MC4R (AG). The presence of the MC4R (A) allele in the animals' genotype contributed to a decrease in fat and solids content of colostrum. The LEPR/HpaII mutation was found to have a considerable effect on the level of most colostrum components (fat, protein and solids) in both pig breeds. Significant decrease in the value of the colostrum components (except lactose) was observed only for animals with the allele LEPR (B). The results obtained suggest that these genes might be used in selection of dam-line pigs as genetic markers of milk quality.

Association between subcutaneous and intramuscular fat content in porcine ham and loin depending on age, breed and FABP3 and LEPR genes transcript abundance.

The objective of the present study was to analyze the level of intramuscular fat (IMF) in loin (musculus longissimus dorsi) and ham (musculus semimembranosus) and the level of subcutaneous fat in these cuts depending on breed, age and the expression level of FABP3 and LEPR genes. The results obtained showed that only the breed influenced on the level of both intramuscular and subcutaneous fat to the same extent (P ≤ 0.001). The age of animals had an effect on fat content of the cuts (P ≤ 0.001) and to a lower extent on the level of IMF in both muscles (P ≤ 0.05). We confirmed highly significant effect of breed and age on the LEPR mRNA abundance--the expression of the this gene increased significantly (P ≤ 0.01) with age and the highest expression was found for the Pulawska breed in m. longissimus dorsi and for the Polish Landrace breed in m. semimembranosus. We observed the high correlations between the transcript level of the LEPR gene and the fat content of individual cuts (P ≤ 0.01). The expression level of FABP3 gene influenced the level of IMF (P ≤ 0.01), but not the level of subcutaneous fat in loin and ham.

Lack of the associations of the polymorphisms in IGF2, MC4R and GNAS genes with reproduction traits in pigs and imprinting analysis of IGF2 gene in ovary and cornus uteri.

In recent years, intensive attention has been put on improving reproductive performance of pigs. Several experiments aimed to identify markers associated with prolificacy, but this issue still remains open. In our study, we investigated associations between polymorphisms in IGF2, GNAS and MC4R genes with reproductive traits of Polish Landrace and Large White pigs. We did not find any significant associations for g. GNAS314T > 324C, IGF2 intron3-g.3072G > A or g. MC4R 1426G > A in Polish Landrace and Large White pigs. In the case of IGF2 intron3-g.3072G > A, this information is of great importance, because this marker is widely implemented in pigs breeding and previous experiments suggested its role in prolificacy of pigs. We also investigated expression of IGF2 gene and showed that this gene is monoallelically expressed in reproductive organs (ovary and cornus uteri).

Frequency of DLK1 c.639C>T polymorphism and the analysis of MEG3/DLK1/PEG11 cluster expression in muscle of swine raised in Poland.

DLK1--(Drosophila like element 1) is a paternally expressed gene, associated with the callipyge phenotype in sheep. In a present study we designed a new real-time PCR alleleic discrimination assay for genotyping of a silent C/T mutation (c.639C>T) in DLK1 gene in swine. The DLK1 c.639C>T mutation was highly polymorphic in all breeds analyzed and C allele was predominant in Landrace and Duroc while T allele was more frequent in Pietrain and Pulawska breed. Moreover, we analyzed mRNA expression of DLK1 and adjacent genes--MEG3 and PEG11 in muscles of swines of different breeds raised in Poland. We did not observe significantly different expression of DLK1, MEG3 or PEG11 mRNA in any of analyzed breeds. We also attempted to assess the effect of DLK1 (c.639C>T) on the expression of genes in callipyge locus but did not find significant differences between animals with alternate genotypes (C/C and T/T homozygotes).

H-FABP and LEPR gene expression profile in skeletal muscles and liver during ontogenesis in various breeds of pigs.

The genes coding for H-FABP (heart acid-binding protein) and LEPR (leptin receptor) are considered to be candidates for lipid metabolism and thus affect fat deposition in pigs. The aim of our study was to assess the amount of H-FABP and LEPR transcript in the skeletal muscles (m. longissimus dorsi, m. semimembranosus) and liver of pigs of various ages. The experiments were carried out on 5 popular breeds of swine raised in Poland which exhibit different levels of fat tissue. Furthermore, we examined the effect of H-FABP and LEPR genotypes (HinfI, HpaII, and HaeIII for H-FABP and HpaII for LEPR) on the expression abundance of these genes. We confirmed a statistically significant relationship between the breed (P<.001), type of tissue (LEPR P<.001; H-FABP P<.01), and age of the animal (P<.05) on the abundance of mRNA transcript of both genes. In all breeds, the expression of the leptin receptor gene increased significantly (P<.01) with age in muscle tissue, whereas this relationship was not observed in liver tissue. However, the expression of the H-FABP gene in muscles did not change with age or breed, although in the liver expression levels were high in young (60 and 90 d) pigs. In conclusion, H-FABP and LEPR genes are strongly related to the development and function of fat tissue in pigs.

Novel porcine housekeeping genes for real-time RT-PCR experiments normalization in adipose tissue: assessment of leptin mRNA quantity in different pig breeds.

The main function of adipose tissue is energy storage and production of various cytokines and hormones, such as leptin. Leptin is a protein hormone synthesized and secreted by adipose tissue. The expression of leptin is strongly dependent on growth and luteinizing hormones, which play an important role in the brain-pituitary axis. The concentration of leptin in blood plasma increases with age and obesity and is associated with the level of leptin mRNA in adipose tissue. Selection of appropriate internal control gene (ICG) for normalization of quantitative PCR data for genes of interest is critical for interpretation of results. The estimation of leptin mRNA is important in the research on regulation of feed intake and metabolic and energy balance. Therefore, the objective of this study was to evaluate the stability of mRNA expression for a number of candidate housekeeping genes in the porcine backfat tissue across different breeds. In our study we used a freeware computer program (geNorm) to evaluate the most stable among eight ICG genes (ß-actin, hypoxanthine phosphoribosyltransferase 1, TATA binding protein, glyceraldehyde-3-phosphate dehydrogenase, ornithine decarboxylase antizyme 1, 60S ribosomal protein L27, 40S ribosomal protein S29, eukaryotic elongation factor (1) in 90 mRNA samples of backfat tissue. In the study we used three breeds differing in muscling: Polish Large White (n=30), Polish Landrace (n=30) and Pietrain (n=30). The results showed that the three most stable genes were ornithine decarboxylase antizyme 1 (OAZ1), 60S ribosomal protein L27 (RPL27) and ß-actin (M=0.579, 0.602 and 0.607, respectively). In order to evaluate the abundance of leptin mRNA, the two most stable genes were used. The highest level of mRNA expression was obtained for PL and the lowest for Pietrain pigs. These results confirmed previous studies which showed that pigs with lean carcass were characterized by a lower level of leptin transcript compared to pigs with large fat deposit. Moreover, we analyzed relationship between C3469T Lep polymorphism and level of leptin mRNA, but did not find significant associations. Our study provides a new panel of housekeeping genes for normalization of the expression of a gene of interest in adipose tissue.

The expression pattern of myogenic regulatory factors MyoD, Myf6 and Pax7 in postnatal porcine skeletal muscles.

The MyoD, Myf6 genes, which belong to the family of muscle regulatory factors (MRFs) play a major role in muscle growth and development. Therefore, they are considered as candidate genes for meat production traits in pigs. These basic helix-loop-helix (bHLH) transcription factors regulate myogenesis: they initiate the formation of muscle fibres and regulate the transcription of muscle specific genes. The paired-box transcription factor Pax7 plays critical roles during fetal development and this protein is essential for renewal and maintenance of muscle stem cells. In particular, expression of Pax7 and MyoD is correlated with presence of active satellite cells, important in hyperplastic and hypertrophic growth in skeletal muscle. The objective of the study was to investigate the level of expression of MyoD, Myf6 and Pax7 genes in porcine skeletal muscles (m. semimembranosus, m. biceps femoris, m. gracilis) in breeds differing in muscularity. Moreover, we investigated expression profile of these genes during ontogenesis in Polish Large White (PLW) and Pietrain pigs in the largest ham muscle (m.semimembranosus). Analysis of several ham muscles showed higher expression of MyoD in the Polish Landrace (PL) breed than in Pietrain and PLW pigs (m. semimembranosus P<0.001; m. biceps femoris P<0.05 and P<0.01, respectively; m. gracilisP<0.01). The level of Pax7 transcript depended on type of muscle and breed. The highest expression was in m. gracilis in Pietrain and the lowest in Polish Landrace. Our results indicate that MyoD and Pax7 genes had higher expression levels in the early stages of development in both investigated breeds. The total expression profile of MyoD and Pax7 genes suggests that higher muscularity in Pietrain pigs is associated with the presence of a greater number of active satellite stem cells compared to other breeds. The expression level of Myf6 gene does not indicate significant differences between muscles, ages and breeds.

Association of the melanocortin-4 receptor (MC4R) with feed intake, growth, fatness and carcass composition in pigs raised in Poland.

The melanocortin-4 receptor (MC4R) is involved in feed intake regulation. A missense mutation G/A in position 1426, which causes Asp-Asn substitution at position 298 in the MC4R gene has been associated with feed intake, fatness and growth, however published results of its effect are inconsistent. In our study we evaluated an effect of missense mutations on carcass composition, growth traits and meat quality in 1191 gilts of five breeds: Polish Large White, Polish Landrace, Pulawska, Pietrain and Duroc. G/A 1426 MC4R mutations were genotyped using a 7500 Real-Time PCR System (Applied Biosystem). Our observations confirm that the G allele is frequent in breeds selected for lean meat content (Pietrain - 92.4%) and rare in breed with a fat carcass (Duroc - 31.5%), which has been previously reported. Our study revealed that the A allele was associated with increased daily feed intake (AA - 2.51kg; GG - 2.31kg in the Pulawska breed, P<0.05), daily gain and backfat thickness (AA - 1.67cm, GG - 1.52cm in PL, P<0.01) and decreased lean meat content (total weight of ham was 0.5kg greater in G/G than in A/A Pulawska animals). We obtained an unexpected result for IMF: higher percentage of IMF was observed in the G/G genotype.