藏猪与杜洛克猪血液中胰岛素样生长因子-1、胰岛素样生长因子结合蛋白-3含量差异研究

为了解国内、外两类猪种肌肉生长发育及血清中相关细胞因子含量差异,以我国特有的藏猪和国外杜洛克猪作为研究对象,采用酶联免疫分析法(ELISA)测定6月龄藏猪、杜洛克猪血清中胰岛素样生长因子-1(IGF-1)、胰岛素样生长因子结合蛋白-3(IGFBP-3)的含量及肌纤维面积(CSA)。结果显示:藏猪血清中IGF-1含量及CSA均极显著低于杜洛克猪(P0.01),IGFBP-3含量极显著高于杜洛克猪(P0.01);CSA与IGF-1含量间呈极显著正相关,与IGFBP-3间呈极显著负相关,说明IGF-1对猪背最长肌生长发育有促进作用,而IGFBP-3可能有抑制作用。     

藏猪与杜洛克猪血液中胰岛素样生长因子-1（IGF-1）含量差异研究

为了解国内、外两类猪种肌肉生长发育及血清中相关细胞因子含量差异,以我国特有的藏猪和国外杜洛克猪作为研究对象,采用酶联免疫分析法（ELISA）测定6月龄藏猪、杜洛克猪血清中胰岛素样生长因子-1（IGF-1）的含量及肌纤维面积。结果显示：藏猪血清中IGF-1含量及肌纤维面积均极显著低于杜洛克猪（P＆lt;0.01）,且两者呈极显著正相关,说明IGF-1是促进猪生长发育特别是背最长肌生长发育的一个重要因素。     

IGF-I与IGFBP-1基因对京海黄鸡生长性状的遗传效应分析

旨在对IGF-I和IGFBP-1基因部分SNPs与鸡生长性状进行关联分析。试验以京海黄鸡为材料,采用PCR-SSCP方法检测2个候选基因的单核苷酸多态性(SNPs),采用一般线性模型(GLM)分析基因型与生长性状的遗传效应。结果表明,IGF-I基因外显子3序列的60bp处有A→G的点突变,在京海黄鸡中检测到AA、AB、BB 3种基因型,A等位基因频率为0.613,B等位基因的频率为0.387;IGFBP-1基因外显子2序列的21bp处有A→T的点突变,104bp处有T→C的突变,在京海黄鸡中检测到CC、CD和DD 3种基因型,C等位基因频率为0.558,D等位基因频率为0.442。IGF-I基因BB基因型个体4周龄体质量显著高于AA和AB型个体(P<0.05);IGFBP-1基因DD基因型个体8周龄体质量显著高于CC基因型个体(P<0.05),4、12和16周龄体质量差异极显著(P<0.01)。因此,推测这些SNPs对京海黄鸡生长性状具有一定的影响,应用于鸡育种过程中的标记辅助选择可以加快育种进程。     

猪胰岛素样生长因子结合蛋白1基因研究进展

胰岛素样生长因子结合蛋白(Insulin-like Growth Factor Binding Proteins,IGF BPs)在调节胰岛素样生长因子(Insulin-likeGrowth Factor,IGF)生理效应中起重要作用,此外,还可以发挥独立于IGF分子网络的作用。IGFBP-1在多种组织和体液中表达,参与机体生长、发育、生殖及血糖等生理过程。本文综述了IGFBP-1的基因定位、结构、作用机制,以及在猪上的研究进展。     

Genetic Effects of IGF-Ⅰ and IGFBP-1 Genes on the Growth Traits in Jinghai Yellow Chicken

旨在对IGF-Ⅰ和IGFBP-1基因部分SNPs与鸡生长性状进行关联分析.试验以京海黄鸡为材料,采用PCR-SSCP方法检测2个候选基因的单核苷酸多态性(SNPs),采用一般线性模型(GLM)分析基因型与生长性状的遗传效应.结果表明,IGF-Ⅰ基因外显子3序列的60 bp处有A→G的点突变,在京海黄鸡中检测到AA、AB、BB 3种基因型,A等位基因频率为0.613,B等位基因的频率为0.387;IGFBP-1基因外显子2序列的21 bp处有A→T的点突变,104 bp处有T→C的突变,在京海黄鸡中检测到CC、CD和DD 3种基因型,C等位基因频率为0.558,D等位基因频率为0.442.IGF-Ⅰ基因BB基因型个体4周龄体质量显著高于AA和AB型个体(P＜0.05);IGFBP-1基因DD基因型个体8周龄体质量显著高于CC基因型个体(P＜0.05),4、12和16周龄体质量差异极显著(P＜0.01).因此,推测这些SNPs对京海黄鸡生长性状具有一定的影响,应用于鸡育种过程中的标记辅助选择可以加快育种进程.     

马鹿茸IGF-1基因5''端克隆及生物信息学分析

对鹿茸IGF-15’端调控序列进行生物信息学分析。提取鲜马鹿茸顶部组织基因组DNA，设计简并引物，利用PCR获得马鹿茸IGF-15’端调控序列及部分外显子（命名为CCS1074），用DNAMAN软件进行CCS1074与其他种属IGF-1基因的序列同源性分析；使用在线分析程序预测CCS1074的潜在转录因子结合位点并比较分析鹿茸区别于其他种属IGF-1的特有潜在转录因子，进行TATAbox、CpG岛、候选启动子区段及信号肽预测。结果获得了包含部分外显子的鹿茸IGF-1基因5’端（Genbank Accession No．DQ234266）；序列同源性分析表明鹿茸IGF-1与牛、羊IGF-1基因5’端存在差异；生物信息学分析表明：在CCS1074180～310base处存在CpG岛，在142～192、238—288、874～924、1359～1409base处可能存在启动子，存在253个潜在的转录因子结合位点；比较发现，ZF5F／ZF5．01和HAND／HAND2-E12．01是鹿茸IGF-1区别于牛、羊、人、猪、家鼠、斑马鱼IGF-1启动子区域的潜在转录因子，这些潜在转录因子与鹿茸生长的关系尚待进一步试验证实；综合分析表明CCS1074〈1～1580base为鹿茸IGF-15’端区域，1581—1966base为外显子1，1903—1966base为信号肽，1967～〉2087base为内含子1。     

六个鸡种IGFBP-3基因第2外显子MspⅠ酶切位点的遗传多态性研究

采用PCR-RFLP和DNA测序技术,研究京海黄鸡、AA鸡、尤溪麻鸡、如皋草鸡、鹿苑鸡、边鸡6个品种IGFBP-3基因第2外显子Msp Ⅰ位点的多态性.结果显示,IGFBP-3基因第2外显子1087 bp处有C→T的突变,鸡群体中表现出AA、AB和BB 3种基因型.3种基因型在6个鸡品种间分布趋势一致:AA为优势基因型,BB为劣势基因型.经x2适合性检验,除京海黄鸡外其余5个品种的Msp Ⅰ位点均处在Hardy-Weinberg平衡状态(P＞0.05).群体遗传分析表明,AA鸡纯合度最高,京海黄鸡纯合度最低.6个鸡品种MspⅠ位点为中度多态.京海黄鸡有效等位基因数最多,为1.900,AA鸡有效等位基因数最少,为1.471.表明鸡IGFBP-3基因在不同品种中具有丰富的单核苷酸多态性,可以进一步作为候选基因来分析其与生长性状的相关性.     

马鹿茸IGF-1基因5′端克隆及生物信息学分析

对鹿茸IGF-1 5′端调控序列进行生物信息学分析。提取鲜马鹿茸顶部组织基因组DNA,设计简并引物,利用PCR获得马鹿茸IGF-1 5′端调控序列及部分外显子(命名为CCS1074),用DNAMAN软件进行CCS1074与其他种属IGF-1基因的序列同源性分析;使用在线分析程序预测CCS1074的潜在转录因子结合位点并比较分析鹿茸区别于其他种属IGF-1的特有潜在转录因子,进行TATA box、CpG岛、候选启动子区段及信号肽预测。结果获得了包含部分外显子的鹿茸IGF-1基因5′端(Genbank Accession No.DQ234266);序列同源性分析表明鹿茸IGF-1与牛、羊IGF-1基因5′端存在差异;生物信息学分析表明:在CCS1074 180~310 base处存在CpG岛,在142~192、238~288、874~924、1359~1409 base处可能存在启动子,存在253个潜在的转录因子结合位点;比较发现,ZF5F/ZF5.01和HAND/HAND2_E12.01是鹿茸IGF-1区别于牛、羊、人、猪、家鼠、斑马鱼IGF-1启动子区域的潜在转录因子,这些潜在转录因子与鹿茸生长的关系尚待进一步试验证实;综合分析表明CCS1 074<1~1580base为鹿茸IGF-1 5′端区域,1581~1966 base为外显子1,1903~1966 base为信号肽,1967~>2087 base为内含子1。     

草原红牛IGFBP3基因多态性及与部分屠宰性状的相关性分析

以草原红牛为研究对象,采用PCR-SSCP方法检测IGFBP3基因的多态性,并将不同基因型与部分屠宰性状进行相关分析.结果表明在第2内含子发现多态性位点,有AA、AB和BB 3种基因型;测序结果显示在8 069 bp处存在C→T的碱基突变;该位点的多态性对草原红牛的眼肌面积有显著影响(P<0.05),AA型显著高于BB型(P<0.05),但与AB型之间差异不显著(P>0.05);其他屠宰性状在不同基因型间差异不显著.由此可以初步推断IGFBP3基因可能是影响草原红牛肉质性状的一个主效基因或是一个与影响肉质性状的QTL连锁的基因.     

鸡IGFBP-2基因的多态性及其与生长、繁殖性状的关系

采用PCR-SSCP方法检测京海黄鸡、AA鸡、尤溪麻鸡、边鸡等4个鸡品种胰岛素样生长因子结合蛋白2（IG-FBP-2）第3内含子部分序列、第4外显子及3′调控区的多态性,并分析其对京海黄鸡生长和繁殖性能的遗传效应。结果显示,IGFBP-2基因在P1、P3引物扩增片段中存在PCR-SSCP多态性。对于P1扩增片段,在京海黄鸡和AA鸡中都检测到EE、EF和FF这3种基因型,在尤溪麻鸡和边鸡中只检测到EE型;测序表明FF型与EE型相比有2处突变（3746T→G,3753CC→TT）;京海黄鸡3种基因型之间300日龄蛋质量、300日龄体质量、出生体质量、8周龄体质量的最小二乘均值差异显著（P〈0.05）。对于P3扩增片段,在4个鸡群体中都检测到GG、GH和HH这3种基因型;测序表明HH型与GG型相比有1处缺失（4415GCGGGAAG）;京海黄鸡3种基因型之间除出生体质量外,其他生长和繁殖性状的最小二乘均值差异均不显著（P〉0.05）。因此,推测IGFBP-2基因可能是影响鸡生长和繁殖的主效基因或与控制该性状的主效基因连锁。     

Change of plasma insulin-like growth factor-1 (IGF-1) concentration with early growth in Japanese beef cattle

The aim of the present study was to examine the change in plasma insulin‐like growth factor‐1 (IGF‐1) concentration with early growth, changes of bodyweight (BW) and relative dairy gain (RDG) in the pre‐ (PRW) and postweaning periods (POW) in Japanese beef cattle, and relationships with metabolites. A total of 33 calves, 22 Japanese black, 6 Japanese shorthorn and 5 of their crossbreed were studied. Insulin‐like growth factor‐1 and metabolite (glucose, triacylglycerol, nonesterified fatty acid) levels in the plasma, from jugular vein blood taken every month, were measured along with BW. Insulin‐like growth factor‐1 in POW increased dramatically with increase of BW (P < 0.05), and the correlation was positive at 0.52 (P < 0.01). Glucose levels correlated significantly with BW, RDG and IGF‐1 (P < 0.01). Metabolic required calorie correlated positively with IGF‐1 (P < 0.01). Also, correlations of BW in POW, with BW and RDG in PRW were positive (P < 0.01). Growth in PRW would be influenced by maternal effects, while active self‐secretion of IGF‐1 in POW might contribute to POW growth. These factors suggested that to increase growth in PRW, maintaining enough maternal effect and IGF‐1 level in POW, was important for establishing better growth after weaning.     

DNA polymorphism of insulin-like growth factor-binding protein-3 gene and its association with birth weight and body weight in cattle

Insulin‐like growth factor‐binding protein‐3 (IGFBP‐3) is a protein that binds the majority of insulin‐like growth factors in circulation for regulation of its action on growth and metabolism of the animals. Animals belonging to Hariana, Holstein‐Friesian (HF) and their crossbreds (HF × Hariana) were studied using polymerase chain reaction‐restriction fragment length polymorphism and nucleotide sequencing of the IGFBP‐3 gene. A 651‐bp fragment of the IGFBP‐3 gene spanning over a part of exon 2, complete intron 2, exon 3 and a part of intron 3 was amplified and digested with HaeIII restriction enzyme. Three patterns of restriction fragments were observed in HF and crossbred cattle revealing polymorphism in both the populations. The frequency of AA, AB and BB genotypes was 0.65, 0.32 and 0.03 in crossbreds and 0.29, 0.65 and 0.06 in HF respectively. The allelic frequency of the A and B allele was 0.81 and 0.19 in crossbreds and 0.62 and 0.38 in HF cattle respectively. Only one restriction pattern (AA genotype) was observed in all the animals of Hariana breed of Bos indicus showing the absence of polymorphism. Nucleotide sequencing revealed a C → A mutation in the intron 2 region of the IGFBP‐3 gene as the cause of the polymorphism. Least squares analysis revealed a significant effect (p < 0.05) of genotypes on birth weight and body weight (weight at 12, 18 and 24 months of age) of the animals. Animals of AB genotype showed higher birth weight and body weight than the animals possessing AA genotype.     

牛、羊胰岛素样生长因子结合蛋白-3基因研究进展

作为胰岛素样生长因子结合蛋白(IGFBPs)中最重要的一类结合蛋白,胰岛素样生长因子结合蛋-白3(IGFBP-3)是血中含量最高、作用最强的结合蛋白。它直接或间接地影响着家畜的泌乳性能、产绒(毛)性状、生长发育及肉品质等生产性状。作者综述了牛、羊IGFBP-3基因多态性及其与泌乳、繁殖和生长发育性状的关系。     

京海黄鸡IGFBP-3基因外显子1和内含子1部分序列多态性及其与生长繁殖性状的相关性

以京海黄鸡母鸡为材料,采用PCR-SSCP技术对JGFBP-3基因外显子1及内含子1部分序列多态位点进行研究,并计算基因型频率、基因频率、卡方值和部分遗传多态性指标.结果表明,在外显子1上没有检测到多态位点,在内含子1上检测到1个多态位点,该位点为中度多态,在160 bp处发生T到G的突变,导致BB、AB型的56日龄体重显著大于AA型(P<0.05),AA型的11月产蛋数显著大于BB、AB型(P<0.05).由此初步推断,内含子1对产蛋及生长发育很可能有一定的促进作用,A为产蛋的优势基因,B为体重的优势基因.     

An insertion/deletion variant of a thymine base in exon 2 of the porcine beta 3‐adrenergic receptor gene associated with loin eye muscle area

An insertion/deletion variant of a thymine base (T5 and T6) in exon 2 of porcine beta 3‐adrenergic receptor (ADRB3) gene has been described. In the current study, we made an association study between the ADRB3 polymorphisms and production traits in 735 Duroc pigs. The allele frequencies for the T5 and T6 alleles in our study population were 0.433 and 0.567, respectively. Any associations between ADRB3 genotype and average daily weight gain during test period, or backfat thickness and intramuscular fat content were not detected in either sex. However the size of the loin eye muscle area (EMA) was significantly associated with ADRB3 genotypes in gilts. T6‐homozygous gilts had a higher mean of EMA (40.6 ± 0.6 cm²) than T5‐homozygous (38.1 ± 0.4 cm², P = 0.002) and heterozygous (38.8 ± 0.3 cm², P = 0.034) gilts. This association was not detected in males. In addition, a multiple traits animal model best linear unbiased predictor (BLUP) analysis revealed that the T6‐homozygous genotype had positive effects on breeding value of EMA. Accordingly, we suggest that ADRB3 polymorphism has the potential to be an important genetic marker for prediction of EMA in Duroc pigs.     

A polymorphism in the insulin-like growth factor 1 gene is associated with postpartum resumption of ovarian cyclicity in Holstein-Friesian cows under grazing conditions

Background

Insulin-like growth factor 1 (IGF-1) gene is considered as a promising candidate for the identification of polymorphisms affecting cattle performance. The objectives of the current study were to determine the association of the single nucleotide polymorphism (SNP) IGF-1/SnaBI with fertility, milk production and body condition traits in Holstein-Friesian dairy cows under grazing conditions.

Methods

Seventy multiparous cows from a commercial herd were genotyped for the SNP IGF-1/SnaBI. Fertility measures evaluated were: interval to commencement of luteal activity (CLA), calving to first service (CFS) and calving to conception (CC) intervals. Milk production and body condition score were also evaluated. The study period extended from 3 wk before calving to the fourth month of lactation.

Results and discussion

Frequencies of the SNP IGF-1/SnaBI alleles A and B were 0.59 and 0.41, respectively. Genotype frequencies were 0.31, 0.54 and 0.14 for AA, AB and BB, respectively. Cows with the AA genotype presented an early CLA and were more likely to resume ovarian cyclicity in the early postpartum than AB and BB ones. No effect of the SNP IGF-1/SnaBI genotype was evidenced on body condition change over the experimental period, suggesting that energy balance is not responsible for the outcome of postpartum ovarian resumption in this study. Traditional fertility measures were not affected by the SNP IGF-1/SnaBI.

Conclusion

To our knowledge this is the first report describing an association of the SNP IGF-1/SnaBI with an endocrine fertility measure like CLA in cattle. Results herein remark the important role of the IGF-1gene in the fertility of dairy cows on early lactation and make the SNP IGF-1/SnaBI an interesting candidate marker for genetic improvement of fertility in dairy cattle.     

北京黑猪FUBP3和USP43基因多态性与眼肌面积性状的关联分析

旨在研究上游远端元件结合蛋白3(far upstream element binding protein 3,FUBP3)及泛素特异性蛋白酶43(ubiquitin specific protease 43,USP43)基因多态性与北京黑猪眼肌面积性状的关系,从而在分子水平指导北京黑猪的育种工作。本研究选取408头北京黑猪收集眼肌面积性状数据并采集肉样提取DNA,根据FUBP3和USP43基因序列设计43对引物进行PCR扩增及测序,运用DNAstar软件分析测序结果,对FUBP3和USP43基因不同基因型与北京黑猪眼肌面积性状进行关联分析并运用荧光定量PCR对两基因进行基因表达差异分析。在FUBP3基因启动子区共有8个突变,其中rs701769847G>A与5~6肋眼肌面积和最后肋眼肌面积均关联显著(P<0.05),rs332131528C>G仅与5~6肋眼肌面积关联显著(P<0.05),rs325550799T>C、rs339464012T>C和rs326069041T>C只与最后肋眼肌面积关联显著(P<0.05);基因表达差异分析表明,rs701769847G>A位点GG型个体的FUBP3基因mRNA水平显著低于AA型个体(P<0.05)。USP43基因启动子区的rs335310752C>T和剪切区的rs323463345G>A均只与最后肋眼肌面积关联显著(P<0.05)。基因表达差异分析表明,rs323463345G>A中GG型个体和AA型个体的USP43基因mRNA水平差异不显著。上述两基因中共有两个位点与5~6肋眼肌面积性状关联显著(P<0.05),6个位点与最后肋眼肌面积性状关联显著(P<0.05),可以作为北京黑猪眼肌面积性状变异的候选基因功能位点,也可以作为潜在的眼肌面积性状分子标记。     

Postpartum follicular and luteal activity in Holstein-Friesian cows genetically selected for high or low mature bodyweight: Relationships with follicle stimulating hormone,insulin, insulin-like growth factor-1 and growth hormone

AIM: To investigate ovarian follicular and luteal activity during the postpartum period of cows genetically selected for high or low mature bodyweight, in relation to metabolic and reproductive endocrine parameters, to determine whether there are differences between strains that could affect fertility outcomes.

METHODS: The presence of follicles ≥5 mm diameter and luteal structures was mapped in the ovaries of 12 high (heavystrain) and 12 low (light-strain) mature bodyweight cows by daily trans-rectal ultrasonography from Day 7 postpartum until the end of their first normal oestrous cycle. Blood samples were collected daily, for measurement of concentrations of follicle stimulating hormone (FSH), progesterone, growth hormone (GH), insulin-like growth factor-1 (IGF-1), and insulin. Intervals to first ovulation were calculated from ultrasonography data.

RESULTS: Heavy-strain cows had shorter intervals than light-strain cows from calving to the emergence of the first (9.0 (SE 0.9) vs 12.4 (SE 1.3) days) and second (16.4 (SE 1.8) vs 20.6 (SE 1.6) days) dominant follicles (p<0.05). Concentrations of FSH in heavy-strain cows prior to the emergence of the second, third and fourth dominant follicles were higher than in light-strain cows (p<0.05). Heavy-strain cows were more likely to have a large (>15 mm diameter) follicle earlier than light-strain cows (p<0.01). Concentrations of insulin and IGF-1, but not those of GH, were higher in heavy- than light-strain cows during the postpartum period (p=0.01 and p=0.02, respectively), and concentrations of both on Day 6 were inversely related to the time of emergence of the first dominant follicle (p>0.01).

Concentrations of progesterone were similar in both strains of cow until Day 10 of the first oestrous cycle, but thereafter were higher in light- than heavy-strain cows until Day 16. Progesterone concentrations in heavy-strain cows declined earlier and more rapidly than in their lighter counterparts.

CONCLUSION: These results indicate that there is a rapid postpartum resumption of follicular activity in both heavy-and light-strain cows, but that there is an earlier emergence of dominant follicles and ovulation in the former. Differences in luteal function, in terms of lower dioestrus progesterone concentrations and an earlier onset of luteolysis, in heavy- than light-strain cows might be sufficient to impair the fertility of the former.     

Circulating concentrations of insulin-like growth factor-1 in dogs with naturally occurring mitral regurgitation

Insulin-like growth factor-1 (IGF-1), which mediates most effects of growth hormone, has effects on cardiac mass and function, and plays an important role in the regulation of vascular tone. In humans, an inverse relationship between degree of heart failure (HF) and circulating IGF-1 concentrations has been found in several studies. In dogs with HF, few studies have focused on IGF-1. We examined circulating IGF-1 concentrations in dogs with mitral regurgitation (MR) caused by myxomatous mitral valve disease. Study 1 included 88 Cavalier King Charles Spaniels (CKCSs) with a broad range of asymptomatic MR (median serum IGF-1: 76.7 microg/L; 25-75 percentile, 59.8-104.9 microg/L). As expected, standard body weight and percentage under- or overweight correlated directly with IGF-1. MR (assessed in 4 different ways) did not correlate with IGF-1. In study 2, 28 dogs with severe MR and stable, treated congestive HF had similar serum IGF-1 concentrations (median, 100.8 g/L; 25-75 percentile, 74.9-156.5 microg/L) as 11 control dogs (79.6 microg/L; 25-75 percentile, 64.1-187.4 microg/L; P = .84). In study 3, the plasma IGF-1 concentration of 15 untreated CKCSs with severe MR was 16.4 +/- 24.2 microg/L lower (P = .02) at the examination when decompensated HF had developed (80.8 +/- 30.9 microg/L) than at a visit 1-12 months earlier (97.2 +/- 39.8 microg/L), possibly in part due to an altered state of nutrition. The studies document that circulating IGF-1 concentrations are not altered before development of congestive HF in dogs with naturally occurring MR, but decrease by approximately 20% with the development of untreated HE In treated HF, circulating IGF-1 concentrations apparently return to within the reference range.