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.     

A melanocortin-4 receptor (MC4R) polymorphism is associated with performance traits in divergently selected Large White pig populations

The melanocortin-4 receptor (MC4R) has a vital role in the control of energy balance and the genetic basis of obesity. A polymorphism, which results in the replacement of aspartic acid with asparagine at position 298 of the porcine MC4R gene, within the seventh transmembrane domain, has previously been described. In the current study, allele frequencies for this Asp298Asn polymorphism were investigated in lines of Large White pigs which had been divergently selected for seven generations based on lean food conversion (LFC), lean growth with ad libitum feeding (LGA), lean growth with restricted feeding (LGS) and daily feed intake (DFI). The association of the Asp298Asn polymorphism with performance traits in these lines was assessed. The frequency of Asp298 was higher (P < 0.001) in the LFC high line (0.48) than the low line (0.00), while the frequency of Asn298 was higher (P < 0.01) in the LGA high line (0.22) than the low line (0.04). When analysed across all lines, the Asp298Asn polymorphism was significantly associated with ultrasonic backfat depth, average daily gain and daily feed intake (P < 0.05). Asp298 homozygous animals had mean values of 13.3 mm, 733 g and 1933 g for backfat, average daily gain and daily feed intake respectively, compared with 14.7 mm, 805 g and 2098 g for Asn298 homozygotes. Therefore, the data support a role for the MC4R Asp298Asn polymorphism in the genetic basis of economically important traits in Large White pigs.     

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.     

Detection of differentially expressed genes in the longissimus dorsi of Northeastern Indigenous and Large White pigs

Recent attention in pig breeding programs has focused on the improvement of pork quality in response to increasing consumer demands. Compared to the fatty-type Northeastern Indigenous (Chinese) breed of pigs, the lean-type Large White has lower intramuscular fat and inferior eating quality from the perspective of the Chinese consumer. In order to investigate the molecular basis of differences in pork quality in Chinese indigenous and Western breeds, longissimus dorsi samples were collected from three adult Northeastern Indigenous and three adult Large White pigs. The RNAs were extracted and hybridized to the porcine Affymetrix GeneChip. Microarray analysis demonstrated differential expression of 1134 genes of which 401 have a known function. One hundred and thirty-six genes were up-regulated and 998 down-regulated in Northeastern Indigenous breed compared to Large White pigs. We screened 10 genes as candidate genes associated with pork quality. We investigated a single nucleotide polymorphism in the 5' regulatory region of the gene FABP4 in 65 Songliao black swine, using PCR-single-strand conformational polymorphism. We found this polymorphism to be highly significantly associated with marbling and intra-muscular fat content (P ≤ 0.01). Genotype BB had higher marbling than AB and AA, but there was no significant difference between AB and AA. Genotype BB and AB had higher intra-muscular fat content than AA, but there was no significant difference between BB and AB. These results help to elucidate the genetic mechanisms behind differences in pork quality and provide a theoretical basis for selection and genetic improvement of meat quality traits in pigs.     

SNPs of MYPN and TTN genes are associated to meat and carcass traits in Italian Large White and Italian Duroc pigs

Myopalladin (MYPN) and titin (TTN) play key roles in skeletal muscle structural organisation and for this reason they may be functional candidates for meat production traits in pigs. We analysed by PCR–RFLP two polymorphisms previously identified and described in the 3′UTR of MYPN and TTN genes in a group of Italian Large White (ILW) and Italian Duroc (ID) pigs with extreme divergent estimated breeding values for some production traits. Significant allele frequency differences were observed for lean cuts (LC) at the MYPN locus in ILW breed and for visible intermuscular fat (VIF) at MYPN and TTN loci in ID breed. Moreover, association analysis between the two genes and some production traits (average daily gain, LC, backfat thickness, ham weight, feed conversion ratio and VIF) was performed in a sample of 409 ILW and 381 ID pigs. On the whole, the results showed significant effects of both genes on some of the considered traits in ID breed, and for TTN gene in ILW animals. These results indicate that both genes can be considered as candidates for meat and carcass traits in ID pigs.     

Variability of the mc1r Gene in Melanic and Non-Melanic Podarcis lilfordi and Podarcis pityusensis from the Balearic Archipelago

The association between polymorphism at the mc1r locus and colour variation was studied in two wall lizard species (Podarcis lilfordi and P. pityusensis) from the Balearic archipelago. Podarcis lilfordi comprises several deep mitochondrial lineages, the oldest of which originated in the Pliocene, while much shallower mitochondrial lineages are found in P. pityusensis. Here, we examined whether specific substitutions were associated with the melanic colouration found in islet populations of these species. Homologous nuclear sequences covering most of the mc1r gene were obtained from 73 individuals from melanic and non-melanic Podarcis from different populations (the entire gene was also sequenced in six selected individuals). MtDNA gene trees were also constructed and used as a framework to assess mc1r diversity. Mc1r showed greater polymorphism in P. lilfordi than in P. pityusensis. However, we observed no substitutions that were common to all melanic individuals across the two species. Only one significant association was detected in the mc1r partial sequence, but this was a synonymous A/G mutation with A alleles being more abundant in melanic populations. In addition, there were no associations between the main dominant phenotypes (green and brown, blue and yellow spots and ventral colour) and synonymous or non-synonymous substitutions in the mc1r gene. There was no statistical evidence of selection on mc1r. This study suggests no relationship between mc1r polymorphism and colour variation in Balearic Podarcis.     

The porcine fat mass and obesity associated (FTO) gene is associated with fat deposition in Italian Duroc pigs

In humans, common variants in the fat mass and obesity associated ( FTO ) gene are associated with body mass index and obesity. Here we sequenced exon 4, parts of introns 3 and 4 and two portions of the 3'-untranslated region of the porcine FTO gene in a panel of nine pigs of different breeds and identified three SNPs. Allele frequencies of the g.276T>G ( AM931150 ) mutation were studied in seven pig breeds. This mutation was used to linkage-map FTO to SSC6. Association analyses between the g.276T>G polymorphism and several traits [pH of semimembranosus muscle and estimated breeding values (EBV) for average daily gain, back fat thickness, lean cuts, ham weight and feed:gain ratio] were carried out in 257 sib-tested Italian Large White pigs. Only feed:gain ratio showed P  <   0.05. A selective genotyping approach was applied, analysing two extreme and divergent groups of Italian Large White pigs selected on the basis of back fat thickness EBV (50 with most positive and 50 with most negative values). Fisher's exact test (two-tailed) was not significant when comparing the allele frequencies of these two groups. The same approach was used in the Italian Duroc breed for which two extreme and divergent groups of animals were selected according to visible intermuscular fat EBV. Differences of allele frequencies between these two groups were highly significant ( P  <   0.00001, P  <   0.001 and P  <   0.0001, considering all animals or only two- or three-generation unrelated animals respectively), indicating association between the analysed FTO marker and intermuscular fat deposition.     

Relationship Among H-FABP Gene Polymorphism, Intramuscular Fat Content, and Adipocyte Lipid Droplet Content in Main Pig Breeds with Different Genotypes in Western China

H-FABP (Heart fatty acid-binding protein), a member of FABP family, plays an essential role in long-chain fatty acid uptake and metabolic homeostasis. Its role in pig intramuscular fat content remains poorly understood, especially in local pig breeds in western China. In this study, the genetic variations of 5′-upstream region and the second intron in porcine H-FABP gene were investigated by PCR-RFLP in 256 pigs including Duroc, Large White, Landrace, Neijiang, Rongchang, Bamei pig, Hanjiang Black, Hanzhong White, and the wild ones. The effect of H-FABP gene on the IMF content was analyzed by the least square method. Lipid droplet morphology and content in adipocytes cultured from pigs with different H-FABP genotypes, were studied by oil red O staining and a triglyceride assay kit. Results showed a Hinf I -RFLP in these eight pig breeds and wild pigs, among which Large white, Bamei pig, Hanjiang Black, Hanzhong White, and wild pigs presented with low polymorphism while the other breeds had intermediate polymorphism. There was no Hae III or Msp I -RFLPs in the four Chinese local pig breeds tested, but Duroc, Landrace, Large White, Hanzhong White and wild pig had polymorphism. Landrace, Large White and wild pigs had low levels of Hae III-and Msp I -RFLP, whereas others had intermediate polymorphism. H-FABP genotypes significantly affected the IMF content (P<0.05). The IMF content ordered by H-FABP genotypes were HH>Hh>hh, DD<Dd<dd, and AA<Aa<aa. The genetic effect values were 3.89, 3.42, 3.17, 2.27, 2.49, 2.91, 2.28, 2.70, and 2.95, respectively. Fat deposition in adipocytes was stronger in the HH, dd and aa genotypes than in others. The results suggest that porcine meat quality may be improved by increasing the frequency of genotype aa-dd-HH in pig breeds.     

Polymorphism of the estrogen receptor 1 locus in populations of pigs of different genotypes and its association with reproductive traits of large white sows

Populations of Large White, Large Black, Poltava Meaty, Pietrian, and Meishan pigs are characterized by intron PvuII polymorphism in the estrogen receptor 1 gene (ESR1). These breeds differ in ESR1 alleles and genotypes frequencies. In the populations, there are deviations in the genotype distribution from the Hardy-Weinberg equilibrium. The established estrogen receptor 1 locus association with the reproductive traits of Large White sows indicates the possibility of using intron ESR1 PvuII polymorphism as a genetic pregnancies marker in pig selection.     

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 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   

Evaluation of the genetic diversity of Laiwu pigs using twenty-seven microsatellite markers

The Laiwu pig, an indigenous pig breed known for extremely high intramuscular fat content, is a well-preserved ancient breed due to long-term natural and artificial selections. In this study, using 27 microsatellite markers jointly recommended by the International Society of Animal Genetics (ISAG) and the Food and Agriculture Organization (FAO), we investigated the genetic diversity of the Laiwu pig breed. The genetic diversity of Laiwu pigs is dramatically low, with the observed heterozygosity ranging from 0.067 to 0.767. Among the 27 microsatellite markers, 10 were high polymorphic loci, 10 were moderate polymorphic loci, six were low polymorphic loci, and no polymorphism was detected at one locus (IGFI). Further analyses with the 10 high polymorphic loci and five moderate polymorphic loci revealed that the Laiwu pig breed was inbred and heterozygous deficient to some extent, but not severely, and that the Laiwu pigs were relatively pure, with almost no hybridization with other breeds. Two subgroups of the current 13 Laiwu pig pedigrees were identified. These results suggest that the Laiwu pig breed has a low diversity and a conservation program must be developed to preserve the “Laiwu pig” gene pool.     

猪ACTA2基因的克隆、表达分析及其与生产性状的关联

为鉴定对猪生产性状有重要影响的新分子标记,文章采用电子克隆结合PCR方法获得了猪肌动蛋白α2(Actin alpha 2, ACTA2)基因编码区序列和部分基因组序列,建立了第2内含子C1554T替换的PCR-HinfⅠ- RFLP基因分型方法,在所检测的7个不同猪群中除大白和梅大群体外,其他群体中均是C等位基因频率高于T等位基因的频率。标记与性状关联分析发现ACTA2基因型与肩部背膘厚、臀部背膘厚、肥肉率、瘦肉率、股二头肌pH和肌内脂肪显著或极显著相关,TT基因型与CC基因型相比具有更高的瘦肉率以及更低的肥肉率和背膘厚。通过Real-time RT-PCR分析发现ACTA2在大白和梅山两个品种猪骨骼肌中的表达量都随着日龄的增加而降低,在各个阶段,梅山猪中的表达量都比大白猪中的表达量高。     

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.     

Copy number variants in Italian Large White pigs detected using high‐density single nucleotide polymorphisms and their association with back fat thickness

The aim of this study was to identify copy number variants (CNVs) in Italian Large White pigs and test them for association with back fat thickness (BFT). Within a population of 12 000 performance‐tested pigs, two groups of animals with extreme and divergent BFT estimated breeding values (EBVs; 147 with negative and 150 with positive EBVs) were genotyped with the Illumina Porcine SNP60 BeadChip. CNVs were detected with penncnv software. We identified a total of 4146 CNV events in 170 copy number variation regions (CNVRs) located on 15 porcine autosomes. Validation of detected CNVRs was carried out (i) by comparing CNVRs already detected by other studies and (ii) by semiquantitative fluorescent multiplex (SQFM) PCR of a few CNVRs. Most of CNVRs detected in Italian Large White pigs (71.2%) were already reported in other pig breeds/populations, and 82.1% of the CNV events detected by penncnv were confirmed by SQFM PCR. For each CNVR, we compared the occurrence of CNV events between the pigs of the high and low BFT EBV tails. Sixteen regions showed significance at P < 0.10, and seven were significant at P < 0.05 but were not significant after Bonferroni correction (Fisher's exact test). These results indicated that CNVs could explain a limited fraction of the genetic variability of fat deposition in Italian Large White pigs. However, it was interesting to note that one of these CNVRs encompassed the ZPLD1 gene. In humans, a rare CNV event including this gene is associated with obesity. Studies identifying CNVs in pigs could assist in elucidating the genetic mechanisms underlying human obesity.     

Association between polymorphisms in cathepsin and cystatin genes with meat production and carcass traits in Italian Duroc pigs: confirmation of the effects of a cathepsin L (CTSL) gene marker

We genotyped single nucleotide polymorphisms (SNPs) in 5 cathepsin or cystatin genes (cathepsin F, CTSF; cathepsin L, CTSL; cathepsin S, CTSS; cathepsin Z, CTSZ; cystatin B, CSTB) in two groups of Italian Duroc pigs: the first group (n. 100) was chosen using a selective genotyping approach with extreme estimated breeding value (EBV) for visible intermuscular fat (VIF); the second group (n. 218) was made of performance-tested Duroc pigs not selected by any criteria. CTSL marker showed a tendency towards association (P < 0.10) with VIF (first group) and back fat thickness (BFT) and average daily gain (ADG; second group). In the second group, the CTSL polymorphism was associated with weight of lean cuts (LC; P < 0.05). Additive effects for all mentioned traits in the second group was significant (P < 0.05). The results we obtained in the Italian Duroc pigs confirmed the results and the direction of the effects already reported for the Italian Large White breed.     

Effects of the MyoG Gene on the Partial Growth Traits in Pigs

Single-nucleotide polymorphisms of the MyoG gene were tested using PCR-SSCP in different pig breeds including Landrace, Large White, Duroc, Shanxi Black, and Mashen pigs, and the effects of the MyoG gene on the birth weight, the weaning weight, the 6-month body weight, and the backfat thickness were also analyzed. On the basis of the published sequence of the porcine MyoG gene, ten pairs of primers were designed, and one polymorphism was found in the PCR product amplified with In2-3 primers. The results showed that: (1) the Landrace, the Large White, and the Duroc breeds differ significantly (P < 0.05) in genotype distribution from the Shanxi Black and the Mashen breeds; (2) On the basis of the fixed effect model, significant differences were found in the birth weight and the backfat thickness among the different MyoG genotypes, whereas no significant differences existed in the weaning weight and the 6-month body weight; (3) Using least square analysis, it was seen that individuals of the BB genotype had significantly less (P < 0.01) birth weight than those of the AA and AB genotypes, with the order being AA>AB>BB; the pigs of the AA genotype had significantly lower (P < 0.01) backfat thickness than those of the AB and BB genotypes, with the order being AA<AB<BB. These results suggest that the genotype has significant effects on the individual birth weight and the backfat thickness, and that the selection of the A allele is favored with regard to the birth weight and the backfat thickness.     

Functional and association studies of the cholesteryl ester transfer protein (CETP) gene in a Wannan Black pig model

Some polymorphisms of the human CETP gene are causally and significantly associated with serum lipids levels; however, the information regarding this gene in pigs is sparse. To evaluate the effects of CETP on blood lipid traits and fat deposition in pig, porcine CETP tissue expression patterns were observed by quantitative real‐time polymerase chain reaction (qPCR) first. High expression was detected in liver, spleen, gluteus medius (GM) muscle and backfat. A de novo polymorphism (AF333037:g.795C>T) in the intron 1 region of porcine CETP was identified. This polymorphism was further genotyped by direct sequencing of the PCR products of 390 Wannan Black pigs, a Chinese native breed population. Association analyses at 45 and 300 days of age revealed highly significant associations between CETP genotypes and serum lipid traits. Furthermore, this polymorphism was proved to be associated with differences in liver CETP mRNA levels: pigs at 300 days of age with the TT genotype had higher levels than did those with other genotypes (P = 0.021). Additionally, analysis at 300 days of age showed that GM CETP mRNA expression correlated positively with serum lipids levels as well as with carcass backfat thickness and intramuscular fat content in GM. These results indicate that CETP is involved in serum, adipose and muscle lipid metabolism in pigs. The mechanisms underlying such relationships and their functional implications are worthy of further research.     

猪ATF4基因多态性与生产性状的关联及基因表达分析

为进一步了解和认识ATF4基因的功能,揭示ATF4对猪脂肪代谢的影响,寻找与肉质性状相关联的分子标记,文章采用PCR方法扩增了ATF4基因部分序列,通过序列比对发现在翻译起始密码子ATG下游159 bp处存在A159G转换,通过PCR-AluⅠ-RFLP对大白猪、长白猪、梅山猪和通城猪进行酶切分型,发现在大白猪和长白猪中均为AA基因型,在梅山猪和通城猪中均为GG基因型。进一步对大白猪×梅山F2群体资源家系进行了酶切分型,并分析该位点的多态性与生产性状的关系。结果表明,ATF4的多态性与臀部平均膘厚存在极显著相关(P<0.01),与胸腰椎间膘厚、平均膘厚、眼肌高、眼肌面积存在显著相关(P<0.05)。采用Real-time PCR分析了ATF4基因在大白猪与梅山猪背最长肌不同发育阶段的表达模式。结果表明,ATF4基因在大白猪和梅山猪胚胎期65 d和出生后3 d中的表达水平相对都比较低,且在两品种间无明显差异;而在出生后60 d和120 d,ATF4基因在大白猪中与梅山猪均出现了上调表达,并且在梅山猪中的相对表达水平要显著高于大白猪。研究结果为进一步深入研究猪ATF4基因在脂肪代谢中的分子机理奠定了基础。