No major effect of the leptin gene polymorphism on porcine production traits

Porcine leptin gene (LEP) and its association with production traits was analysed in the Polish Large White (PLW, n = 135), Polish Landrace (PL, n = 120) and synthetic line 990 (L990, n = 184). Two fragments of exon 3 of LEP were studied with the use of RFLP and SSCP techniques. The frequencies of C allele for the T3469C polymorphic site were 0.11, 0.10 and 0.11 in PLW, PL and L990, respectively. Phenotypic data were collected for the average daily weight gain, the feed conversion ratio, the weight of abdominal fat, backfat thickness (five measurements), intramuscular fat, meat content, loin weight, loin muscle area, ham weight and ham cut weight. The contrasts between TT and TC genotypes at the T3469C polymorphic site were estimated in the unitrait Animal Model with genotype at the RYR1 locus included. The lowest p values occurred for association test between T3469C polymorphism and intramuscular fat content in PLW (0.26 ± 0.14%, p = 0.05) and loin weight in L990 (?0.32 ± 0.13 kg, p = 0.01). We conclude that the detected associations are population‐specific and the analysed polymorphism of the LEP gene does not contribute directly to genetic variability of growth and carcass traits in pigs.     

The Association Analysis Between the SNPs of Leptin Gene and Litter Size Trait in Pigs

The research was aimed to analyze the association of Leptingene and breed traits on pigs. Three pairs of primers were designed which based on DNA sequence of pig Leptin gene (GenBank accession No.U66254, AF026976),the single nucleotide polymorphisms (SNPs) of Leptin gene were detected by PCR-SSCP in Sujiang and Jiangquhai pigs. The results showed that three genotypes of AA,AB and BB were identified, and the mutation (T3469C) in CDS of Leptin gene was found,but amino acid change weren't happened. The Chi-square test results showed that the Leptin genotype distribution of Sujiang and Jiangquhai pigs was in Hardy-Weinberg equilibrium (P>0.05). The polymorphism information content (PIC) were moderate polymorphism in Sujiang pigs,and low polymorphism in Jiangquhai pigs. The statistic analysis results showed that there were no significant association between the SNP of Leptin genotype and litter size trait(P>0.05). But the average litter size of T3469C site AB genotype was higher than AA and BB genotypes, it was concluded that the T3469C polymorphism of Leptingene might be the genetic marker which potentially affected pig breed traits.     

猪Leptin基因多态性及其与产仔性状的相关性分析

试验旨在探究Leptin基因对猪产仔性状的影响,为其新品种持续选育提供遗传学依据。参考GenBank发布的猪Leptin基因DNA序列（登录号:U66254、AF026976）设计3对引物,采用PCR-SSCP技术分析Leptin基因在苏姜猪和姜曲海猪中的单核苷酸多态性（SNP）,并用统计学方法分析基因座多态性与产仔性状的相关性。结果发现,Leptin基因在苏姜猪、姜曲海猪群体中均具有多态性,检测到3种基因型:AA、AB和BB;测序结果发现,编码区第3 469位碱基处发生单碱基突变（T3469C）,为同义突变。χ²适合性检验显示,2个猪种Leptin基因型分布处于Hardy-Weinberg平衡（P>0.05）。多态信息含量（PIC）分析显示,苏姜猪为中度多态,姜曲海猪为低度多态。Leptin基因型与产仔性状关联分析显示,不同基因型间产仔性状差异不显著（P>0.05）,但T3469C位点AB基因型平均产仔数较AA和BB基因型高,初步推断Leptin基因T3469C位点可能是影响猪产仔性状潜在的遗传标记,建议做进一步的研究。     

Associations of leptin gene polymorphisms with production traits in pigs

The associations of leptin (LEP) gene polymorphisms C798T, T2411C, T3266G and T3469C with production traits were investigated in a F2 pig population produced by divergent crosses. The statistical model included genotype, sex, batch and genotype by sex interaction as fixed effects and sire as random effect. Polymorphism C798T was associated with variation in total teat number (p < 0.02) and left teat number (p < 0.03), and polymorphism T3469C was associated with weight at 21 days (p < 0.03), 42 days (p < 0.05), 63 days (p < 0.02) and 77 days of age (p < 0.04) as well as feed intake (p < 0.01), average daily gain (p < 0.01), feed conversion (p < 0.01), bacon depth (p < 0.03) and slaughter weight (p < 0.03). Phenotypic associations were also performed by combining T3469C and C798T genotypes. Interaction between C798T genotypes and sex was observed for some traits. LEP genotypes had significant influence on performance traits, and can be considered as potential genetic markers for selection. However, these results have to be validated in commercial herds.     

An age-dependent association between a leptin C3469T single nucleotide polymorphism and intramuscular fat content in pigs

The objective of this work was to investigate whether the C3469T mutation at exon 3 of the pig leptin (Lep) gene is associated with fatness and if the magnitude and direction of these associations is consistent within age. Repeated measurements on body weight (BW), ultrasonic subcutaneous backfat (SB) and loin-muscle (LT) thickness, and intramuscular fat (IMF) content and composition were taken at 160, 180, 215 and 225 days of age on 162 Duroc barrows previously genotyped for the C3469T mutation. Plasma leptin concentration was determined at 180 days. A total of 505 records of IMF content and composition, which were determined both in live samples of longisimus dorsi muscle (LM) and in carcass samples of LM and gluteus medius muscle, were used in the analyses. The genotypic frequencies associated with the C3469T Lep mutation were 0.76, 0.23, and 0.01 for TT, TC, and CC genotypes, respectively. The effect of the Lep genotypes was assessed using a Bayesian approach based on a polynomial mixed model on age. No evidence was found for an association of the Lep genotypes neither with BW, SB thickness nor with plasma leptin concentration. The posterior mean of the difference TT–TC ranged, for LT, from 0.82 (160 days, posterior probability density (PP)> 0 of 87%) to 1.98 mm (220 days, PP > 0 of 96%) and, for IMF, from 0.96 (160 days, PP > 0 of 84%) to − 1.64 mg/g DM (220 days, PP > 0 of 2%). The results suggest an age-dependent association of the C3469T Lep polymorphism with LT and IMF, which may be useful to explain some controversial results in the literature concerning the association of this polymorphism with IMF content.     

Leptin基因对陆川猪和大白猪产仔数的影响

根据leptin基因在GenBank中的已知序列设计两对引物,采用PCR-SSCP技术在陆川猪和大白猪群体中进行单核苷酸多态性（SNPs）检测,发现了两个SNPs位点,并对其不同基因型个体PCR回收产物进行测序,同时对这两个SNPs位点与产仔数进行了关联分析。结果表明：这两个SNPs位点分别是在第3 469碱基处发生了T→C突变和第3 714碱基处发生了T→G突变,这两个突变都是同义突变;T3714G SNP对陆川猪产仔数影响显著（P〈0.05）,而对大白猪产仔数影响不显著（P〉0.05）,T3 469C SNP对陆川猪和大白猪产仔数影响均不显著（P〉0.05）。     

藏猪EGLN1基因exon 4及部分内含子变异研究

青藏高原的高寒低氧极端环境影响动物的正常生存和物种繁衍,藏猪在长期的适应性驯化中对低氧环境有着更强的适应能力。本研究对高海拔猪种藏猪(甘孜藏猪、迪庆藏猪、西藏藏猪和合作藏猪)和八眉猪以及低海拔猪种烟台黑猪、长白猪和大白猪EGLN1基因exon 4及部分内含子的核苷酸序列进行测定和分析。结果表明:在EGLN1基因exon 4序列的266bp处发生了C→T的颠换,藏猪和八眉猪在该位点均为C,而烟台黑猪和引入的选育猪种长白猪、大白猪在该位点出现了C/T杂合现象;基于变异位点的等位基因C和T共界定了3种基因型(CC、CT、TT),C和CC分别是高海拔地方猪种藏猪和八眉猪的唯一等位基因和基因型;而在低海拔选育猪种中,CC基因型频率仅为0.20,其他2种基因型均为0.40,T为优势等位基因,频率达到0.60,而等位基因C的频率仅为0.40。本研究认为,高海拔猪种(2 300 m以上)藏猪和八眉猪与低海拔猪种烟台黑猪、长白猪和大白猪在EGLN1基因上存在特征性遗传变异,根据等位基因(C和T)及基因型(CC、CT、TT)在海拔上的特征差异,推测可能与不同地理区域条件下的长期适应性驯化有关。     

Polymorphism of IGFBP2 Gene and Its Association with Meat Quality Traits in Pigs

This study was aimed to analyze the genetic variation of insulin-like growth factor binding protein 2 (IGFBP2) gene effect on meat quality traits in pigs. A pair of primer was designed,which based on DNA sequence of IGFBP2 gene (GenBank accession No.:BV727778). The genetic polymorphism of IGFBP2 gene was detected by PCR-RFLP in Sujiang pig, Jiangquhai pig and Landrace pig, and the association between IGFBP2 gene polymorphism and meat quality traits in three pig breeds was analyzed. The results showed that three genotypes (AA, AB and BB) were detected. The Chi-square test results showed that the genotype distribution of three pig breeds was in Hardy-Weinberg equilibrium (P > 0.05). The polymorphism information content (PIC) were moderately polymorphic in three pig breeds. According to the association analysis of different genotypes, significant difference was found in color value, intramuscular fat content and marbling, the color value (except for Jianquhai pig) and intramuscular fat content of AA genotype were significantly higher than AB and BB genotypes (P < 0.05), the marbling of AA and AB genotypes was significantly higher than BB genotype (P < 0.05). In conclusion,the polymorphism of IGFBP2 gene intron 3 had significant effects on meat quality traits, and was candidate gene for the potential impact on meat quality traits of pigs.     

猪IGFBP2基因遗传多态性及其与肉质性状的关联分析

试验旨在探究胰岛素样生长因子结合蛋白2（insulin-like growth factor binding protein 2,IGFBP2）基因遗传变异对猪肉质性状的影响,为猪品种选育提供遗传学依据。参考GenBank中猪IGFBP2基因序列（登录号：BV727778）设计1对引物,采用PCR-RFLP技术检测IGFBP2基因在苏姜猪、姜曲海猪、长白猪中的遗传多态性,并分析基因座多态性与猪肉质性状的相关性。结果发现,3个猪种IGFBP2基因内含子3均具有Msp Ⅰ酶切多态性,分别检测到3种基因型：AA、AB和BB;χ²适合性检验显示,3个猪种IGFBP2基因基因型分布均处于Hardy-Weinberg平衡状态（P > 0.05）;多态信息含量（PIC）分析显示,3个猪种均为中度多态。IGFBP2基因不同基因型与肉质性状关联分析显示,3个猪种不同基因型肉色、肌内脂肪含量、大理石纹差异显著,AA基因型肉色（姜曲海猪除外）、肌内脂肪含量均显著高于AB、BB基因型（P < 0.05）;AA、AB基因型大理石纹显著高于BB基因型（P < 0.05）,其余指标间差异不显著（P > 0.05）。综上,苏姜猪、姜曲海猪、长白猪IGFBP2基因内含子3多态性对肉质性状影响显著,可作为潜在影响猪肉质性状的候选基因。     

Genetic relationships of body composition, serum leptin, and age at puberty in gilts

Leptin produced by adipocytes acts through leptin receptors in the hypothalamus to control appetite and food intake and thus communicates information about degree of fatness. It is thought that a degree of body fat is required for initiation of puberty and maintenance of reproductive function in mammals. The objective of this study was to determine whether polymorphisms in the leptin (LEP), leptin receptor (LEPR), paired box 5 (PAX5), aldo-keto reductase (AKR), and pro-opiomelanocortin (POMC) genes were associated with age, leptin concentration, backfat as an indicator of body condition, or BW at puberty in 3 lines of gilts and to characterize genetic relationships among these traits. The first 2 lines, born in 2001, were formed by crossing maternal White Cross (Yorkshire x Maternal Landrace) gilts to Duroc (n = 210) or (lean) Landrace (n = 207) boars. The remaining line (n = 507), born in 2002, was formed by crossing progeny of the Duroc- and Landrace-sired lines. At first estrus, age, BW (BWP), and backfat (BFP) at puberty were recorded and blood was collected for leptin assays. Nine SNP were detected in candidate genes/regions: 1 in LEP, 3 in LEPR, 1 in PAX5, 2 in AKR, and 2 in POMC. Animals were genotyped for each of the SNP; genotypes were validated using GenoProb. The association model included fixed effects of farrowing group, covariates of SNP genotypic probabilities (from GenoProb), and random additive polygenic effects to account for genetic similarities between animals not explained by SNP. Variance components for polygenic effects and error were estimated using MTDFREML. Leptin concentrations were logarithmically transformed for data analysis. All 4 traits were moderately to highly heritable (0.38 to 0.48). Age and leptin at puberty had a significant (P < 0.01) genetic correlation at -0.63 +/- 0.097, and the genetic correlation between BWP and age at puberty was 0.65 +/- 0.083 (P < 0.01). Significant additive associations (a; P < 0.05) were detected at PAX5 for age at puberty (a = 3.2 d) and for BFP (a = 0.61 mm). One SNP in LEPR was associated with leptin concentration (a = 0.31 log units; P < 0.05). The associations from PAX5 correspond to a QTL peak for age at puberty detected on SSC1. Although not necessarily the causative mutation, this result implies that a QTL that can decrease age at puberty without increasing BFP and BWP at puberty may exist in this region in commercial pigs.     

Effects of porcine leptin receptor gene polymorphisms on backfat thickness,fat area ratios by image analysis,and serum leptin concentrations in a Duroc purebred population

The leptin receptor (LEPR) gene is considered a candidate gene for fatness traits. It is located on SSC 6 in a region in which quantitative trait loci (QTLs) for backfat thickness (BF), fat area ratios, and serum leptin concentration (LEPC) have previously been detected in a Duroc purebred population. The objectives of the present study were to identify porcine LEPR polymorphisms and examine the effects of LEPR polymorphisms on fatness traits in this same population. The Duroc pigs (226 to 953 pigs) were evaluated for BF, fat area ratios using image analysis, and LEPC. A total of seven single nucleotide polymorphisms (SNPs) in the full‐length LEPR coding region were identified in pigs from the base population. Four non‐synonymous SNPs of the LEPR gene and 15 microsatellite markers on SSC 6 were then genotyped in all pigs. During candidate gene analysis, we detected significant effects of the non‐synonymous SNP c.2002C>T in exon 14 on all traits. In fine mapping analysis, significant QTLs for BF, fat area ratios, and LEPC were detected near the LEPR gene in the same region. These results indicated that the c.2002C>T SNP of LEPR has a strong effect on BF, fat area ratios and LEPC.     

猪PRLR和FSHβ基因多态性检测及其与繁殖性状的关联分析

试验旨在研究催乳素受体（prolactin receptor,PRLR）和卵泡刺激素β（follicle-stimulating hormone β,FSHβ）基因的多态性及其与母猪繁殖性状的关联分析。采用PCR-RFLP方法对大白猪、杜洛克猪和长白猪3个猪种共487头母猪进行了PRLR和FSHβ基因多态性检验,并采用单因子方差分析、LSD法分析不同基因型与母猪总产仔数、产活仔数、断奶窝重和断奶窝仔数等繁殖性状的相关性。结果表明,PRLR基因在大白猪和长白猪中B等位基因均为优势基因,基因频率分别为0.717和0.548,大白猪BB基因型总产仔数显著高于AA基因型（P<0.05）,长白猪BB基因型断奶窝重显著高于AB基因型（P<0.05）;FSHβ基因在大白猪、杜洛克猪和长白猪3个猪种中B等位基因均为优势基因,基因频率分别为0.804、0.760和0.789,长白猪BB基因型总产仔数和产活仔数均显著高于AB基因型（P<0.05）,呈现BB>AA>AB趋势。因此,PRLR和FSHβ基因对荣昌猪场母猪繁殖性能有一定影响,可作为母猪繁殖性能分子选育的候选参考基因。     

猪蛋白酪氨酸磷酸酶-1B基因第7内含子多态性与猪脂肪沉积性状的关联性

本研究旨在分析蛋白酪氨酸磷酸酶-1B(PTP-1B)基因部分SNP与猪脂肪沉积性状的关联性。选取国外瘦肉型猪种大约克夏16头、长白猪21头和中国脂肪型地方猪种梅山猪22头以及"大白×梅山"F2代资源家系213头为研究材料,利用PCR-Eco88I-RFLP方法检测PTP-1B基因第7内含子序列存在G/A突变的多态性,并进行方差分析。结果表明,大白和长白猪种为单一的A等位基因,而梅山猪种以杂合子GA基因型为主,对213头"大白×梅山"F2代资源家系进行性状关联分析,该位点与猪板油质量、内脂率、胸腰椎间背膘厚等脂肪沉积性状显著相关(P<0.05),且杂合子GA基因型个体的板油质量和内酯率都最低(GA基因型个体相似文献   

Hormonal regulation of leptin receptor expression in primary cultures of porcine hepatocytes

A study was conducted to elucidate hormonal control of leptin receptor gene expression in primary cultures of porcine hepatocytes. Hepatocytes were isolated from pigs (52 kg) and seeded into collagen-coated T-25 flasks. Monolayer cultures were established in medium containing fetal bovine serum for 1 day and switched to a serum-free medium for the remainder of the 3-day culture period. To establish basal conditions hepatocytes were maintained in serum-free William's E medium containing 10 nM dexamethasone and 1 ng/ml insulin. For the final 24 h, insulin (1 or 100 ng/ml) or glucagon (100 ng/ml), were added in the presence or absence of 100 nM triiodothyronine (T3). RNA was extracted and quantitative RT-PCR was performed with primers specific for the long form and total porcine leptin receptors. Leptin receptor expression was calculated relative to co-amplified 18S rRNA. Expression of the long form of the leptin receptor was confirmed under basal conditions. Insulin, glucagon and synthetic human proteins (ghrelin and GLP-1) at 100 ng/ml had no influence on leptin receptor expression; the addition of T3 was associated with a marked increase (P < 0.001) in expression of total and long forms of the leptin receptor by 1.6 and 2.4-fold, respectively. Addition of leptin to cells which were pre-treated with T3 for 24 h (to up-regulate leptin receptor expression), confirmed the lack of a direct effect of leptin on glucagon-induced glycogen turnover and cAMP production. These data suggest that porcine hepatocytes may be insensitive to leptin stimulation even when leptin receptor expression is enhanced by T3.     

Prepuberal evolution of plasma leptin levels in gilts of thrifty genotype (Iberian pig) and lean commercial crosses (Large White × Landrace)

Current study aimed to determine possible differences in plasma leptin levels during the prepuberal period and their relationship with the onset of puberty in gilts of obese thrifty genotype (Iberian breed) and lean genotype (Large White × Landrace commercial crosses) reared under similar conditions. Plasma leptin concentration increased linearly during the 7 weeks prior to the day of puberty attainment in both genotypes (P<0.005, r=0.707 for LW × L and P<0.0005, r=0.874 for Iberian gilts). However, leptin levels in the Iberian gilts was higher from the first sample of the experimental period, with females having 16 weeks-old (2.7±0.3 vs 1.7±0.2 ng/ml in LW × L; P<0.001), to the onset of puberty (8.5±0.7 vs 2.8±0.3 ng/ml in LW × L; P<0.005). Thus, the current study reinforces previous data on changes in around puberty and evidences, for the first time, profound differences in prepuberal plasma leptin levels between gilts of obese (Iberian) and lean genotypes (LW×L).     

松辽黑猪和长白猪催乳素受体基因第10外显子多态性与繁殖性状的关联分析

试验旨在探讨催乳素受体(prolactin receptor,PRLR)基因遗传多态性及其与松辽黑猪、长白猪繁殖性状的关联性。采用PCR-RFLP方法对松辽黑猪和长白猪PRLR基因第10外显子进行了多态性检测,并运用最小二乘法分析各基因型对母猪繁殖性能的遗传效应。结果表明,两个猪种中都存在A、B 2个等位基因和AA、AB、BB 3种基因型,松辽黑猪和长白猪等位基因A的频率分别0.551和0.578,A为优势基因。两个猪种的AA基因型个体的断奶仔猪数、乳头数都显著高于其他两个基因型(P＜0.05),初产松辽黑猪AA基因型个体的TNB、NBA显著高于其他基因型(P＜0.05),对其他繁殖性状影响不显著,但呈现出AA＞AB＞BB的趋势。     

DHRS4基因在猪骨骼肌中的表达及其与生长性状的关联分析

为探讨猪DHRS4基因在骨骼肌中的表达及其与生长性状的相关性,本研究分析了DHRS4基因在长白猪出生后30、180和300 d不同类型骨骼肌中的表达,并在蓝思白猪资源群体中筛选DHRS4基因的多态性位点,进一步通过Sequenom SNP分型技术对DHRS4基因进行基因分型,分析了DHRS4基因多态性位点与猪生长性状（料重比、平均日增重和平均背膘厚）的相关性。结果显示,DHRS4基因在30、180和300 d长白猪的胸肌、腿肌和背肌中均有表达,在胸肌和腿肌中,DHRS4基因在30 d的表达显著或极显著高于180和300 d（P<0.05;P<0.01）,而在背肌的不同发育时期中,其表达水平没有显著差异（P>0.05）。此外,本研究在蓝思白猪资源群体中找到了3个位于DHRS4基因上的单核苷酸多态性位点：rs334250151、rs342446613和rs326982309。关联分析结果表明,rs334250151位点与料重比呈显著相关,该位点AA基因型个体的料重比显著低于TT基因型个体（P<0.05）。研究揭示,DHRS4基因可能参与猪的骨骼肌发育,rs334250151位点可以作为一个新的分子标记应用于猪生长性状的遗传改良中。     

Patterns of corpora lutea growth and progesterone secretion in sows with thrifty genotype and leptin resistance due to leptin receptor gene polymorphisms (Iberian pig)

The current study aimed to compare luteal function, as measured by corpora lutea dynamics and progesterone secretion, in 10 sows with obesity/leptin resistance genotype (Iberian pig) and 10 females of lean commercial crosses (Large White × Landrace). In all the animals, the oestrous cycle was synchronized with progestagens, and ovulation was induced by exogenous gonadotrophins. Thereafter, number and size of follicles and plasma oestradiol concentration were determined at oestrus detection, and number and size of corpora lutea and progesterone concentration were evaluated from Day 4 to 12 of the cycle. There were no differences between genotypes in follicle population and oestradiol concentration, and ovulation rate (15.2±1.3 in Iberian vs 12.7±1.8 in LWxL sows); however, there was a higher percentage of Iberian than control sows showing luteal cysts (66.7% vs 30%, respectively; p<0.05). In both breeds, both total luteal area and plasma progesterone concentration grew linearly from Day 4 to 8 (p<0.01) and remained more stable between Days 8 and 12, without significant differences between genotypes. In conclusion, current study supports that ovulatory processes and luteal functionality are not the main limiting factors for prolificacy in a pig model of leptin resistance and obesity.     

Single- and joint-population analyses of two experimental pig crosses to confirm quantitative trait loci on Sus scrofa chromosome 6 and leptin receptor effects on fatness and growth traits

The primary goal of this study was to detect and confirm QTL on SSC6 for growth and fatness traits in 2 experimental F(2) intercrosses: Iberian x Landrace (IB x LR) and Iberian x Meishan (IB x MS), which were used in this study for the first time in a QTL analysis related to productive traits. For this purpose, single- and joint-population analyses with single and bivariate trait models of both populations were performed. The presence of the SSC6 QTL for backfat thickness previously identified in the IB x LR cross was detected in this population with additional molecular information, but also was confirmed in the IB x MS cross. In addition, a QTL affecting BW was detected in both crosses in a similar position to the QTL detected for backfat thickness. This is the first study in which a QTL affecting BW is detected on SSC6 in the IB x LR cross, as well as in the IB x MS resource population. Furthermore, we analyzed a previously described nonsynonymous leptin receptor (LEPR) SNP located in exon 14 (c.2002C > T) for causality with respect to this QTL within both F(2) populations. Our results supported the previously reported association between LEPR alleles and backfat thickness in the IB x LR cross, and this association was also confirmed within the IB x MS cross. An association not reported before between LEPR alleles and BW was identified in both populations.