摘除卵巢对母羊不同组织中MSTN、MYOG基因表达量的影响

试验通过研究卵巢摘除对布尔山羊杂种母羊羔肝脏、背最长肌、股二头肌中肌细胞生长抑制素(MSTN)和肌细胞生成素(MYOG)基因表达量的影响,旨在探讨去卵巢是否可以促进羔羊生长发育。将体重相近、长势良好的5月龄布尔山羊杂种母羊羔,随机分为处理组(n=20)和对照组(n=20)。试验初期摘除处理组母羊羔卵巢,对照组不摘除。饲养60d后,利用实时定量PCR法检测肝脏、背最长肌、股二头肌中MSTN和MYOG基因mRNA相对表达量。结果表明,卵巢摘除后,母羊股二头肌中MSTN基因mRNA相对表达量是对照组的8.73倍,差异极显著(P0.01),但背最长肌和肝脏中MSTN基因mRNA水平低于对照组(P0.05);处理组背最长肌和股二头肌中MYOG基因表达量极显著高于对照组的5.12倍和6.51倍(P0.01),肝脏中MYOG基因表达量为对照组的2.23倍(0.01P0.05)。可见,卵巢摘除可以上调背最长肌、股二头肌和肝脏中MYOG基因的表达以及股二头肌中MSTN基因的表达。     

不同物种肌生成素(MyoG)密码子使用模式分析

肌生成素(MyoG)是最重要的肌生成调节因子(MRFs)之一,是控制骨骼肌生成的关键调控因子。密码子使用偏倚(codon usage bias, CUB)是目前基因组特有的一种特征,可以揭示物种间的特异性差异。此外,在基因序列密码子使用模型中,碱基组成的动态变化可以更好地了解某一基因的分子机制和进化关系。本研究从GenBank数据库中选取8个物种的MYOG编码序列,用R软件计算其核苷酸组成(GC含量)和遗传指标,包括有效密码子数(ENC)、相对同义密码子使用(RSCU)和相对密码子使用偏量(RCBS)。然后,对不同物种MYOG密码子的密码子偏好性和碱基组成的动态进行了分析比较。结果表明,不同物种的MYOG密码子使用受GC偏置的影响,尤其是GC3s偏置。最优密码子偏倚以A/U结束。CUG、GUG、AUG、AGC和GUU在不同物种中均表现出较高的RSCU值,是最优密码子。人与鸡的密码子功能性质相似,牛与羊的密码子功能性质相似。ENC值与GC3s呈极显著负相关(P<0.01)。最优密码子频率的FOP值越高,说明不同物种的MYOG在密码子使用上存在较大的偏性。     

小鼠miR-487b-3p对C2C12增殖和分化调控作用的研究

旨在研究miR-487b-3p对小鼠C2C12细胞增殖与分化的具体调控机制。本研究以小鼠成肌细胞系(C2C12)为研究对象,利用qRT-PCR检测miR-487b-3p在小鼠各组织和C2C12细胞增殖与分化过程中的表达情况;通过转染miR-487b-3p mimics和miR-487b-3p inhibitor后,利用qRT-PCR检测转染效率,并采用qRT-PCR和Western blotting检测增殖标记基因PCNA和分化标记基因MYOD、MYOG和MYHC的表达差异;利用生物信息学软件对miR-487b-3p靶基因进行预测,利用qRT-PCR检测小鼠C2C12细胞增殖与分化过程中PITX2的表达情况,并检测过表达(抑制)miR-487b-3p后PITX2的表达差异,通过双荧光素酶报告系统验证miR-487b-3p和PITX2的靶标关系。结果表明,miR-487b-3p在小鼠骨骼肌中相对表达最高,在C2C12细胞增殖与分化的过程中miR-487b-3p的表达量整体上呈现上升趋势,推测miR-487b-3p参与调控骨骼肌发育。过表达miR-487b-3p后,极显著上调miR-487b-3p的表达(P0.01),在转录和蛋白水平均极显著上调PCNA的表达(P0.01),显著下调MYHC、MYOD和MYOG基因的表达(P0.05),同时显著下调MYHC(P0.05)、MYOD(P0.01)和MYOG蛋白(P0.05)的表达;而抑制miR-487b-3p后,极显著下调miR-487b-3p的表达(P0.01),显著下调PCNA基因的表达(P0.05),极显著下调PCNA蛋白的表达(P0.01),显著上调MYHC、MYOD和MYOG蛋白的表达(P0.05),上调MYHC、MYOD和MYOG基因的表达,但未达显著水平(P0.05)。在C2C12细胞增殖和分化过程中,PITX2与miR-487b-3p表达趋势相反;过表达miR-487b-3p能极显著下调PITX2mRNA的表达(P0.01),相反,抑制miR-487b-3p的表达则极显著上调PITX2mRNA的表达(P0.01);双荧光素酶报告系统验证PITX2是miR-487b-3p的直接靶基因。综上表明,miR-487b-3p通过靶向PITX2促进C2C12细胞增殖而抑制其分化。     

单核苷酸多态性及其在猪育种中的应用

单核苷酸多态性是一类在基因组中最普遍存在的分子标记。本文综述了单核苷酸多态性的概念、特点、检测技术及猪生长性状、背膘厚和瘦肉率、肉质性状和抗病性状中单核苷酸多态性的应用。     

共轭亚油酸对C2C12肌细胞生脂和生肌分化的影响

本研究分析了共轭亚油酸(CLA)对C2C12肌细胞生脂转分化和生肌分化的影响。分别培养并诱导C2C12鼠源肌细胞生脂转分化和正常的生肌分化,同时分别使用终浓度为50μmol/L的c9,t11-CLA和t10,c12-CLA处理细胞,并设对照组,取生脂转分化第10天和生肌分化第8天的细胞用于实时定量PCR检测,观察c9,t11-CLA和t10,c12-CLA对C2C12肌细胞不同分化的影响。结果表明:1)与对照组相比,c9,t11-CLA促进了C2C12肌细胞的生脂转分化,显著增加了细胞内甘油三酯(TG)含量(P0.05),显著上调了细胞内脂肪酸合成酶(FAS)、CCAAT增强子结合蛋白α(C/EBPα)、过氧化物酶体增殖剂激活受体γ(PPARγ)和脂肪酸结合蛋白4(FABP4)基因的表达水平(P0.05);与对照组相比,t10,c12-CLA则抑制了C2C12肌细胞的生脂转分化,显著减少了细胞内TG含量(P0.05),显著下调了细胞内C/EBPα、PPARγ和FA BP4基因的表达水平(P0.05)。免疫印迹杂交结果显示FAS和FABP4的蛋白质表达水平也发生了与基因表达相一致的变化。2)与对照组相比,t10,c12-CLA抑制了C2C12肌细胞的生肌分化,显著减少了细胞内肌管数/细胞数(P0.05),显著下调了细胞内肌细胞生成素(MYOG)和成肌分化抗原(MYOD)基因的表达水平(P0.05);与对照组相比,c9,t11-CLA则显著上调了细胞内MYOG基因的表达水平(P0.05),对C2C12肌细胞的生肌分化有一定程度的促进作用。免疫印迹杂交结果显示MYOG和MYOD的蛋白质表达水平也发生了与基因表达相一致的变化。以上结果表明,CLA对动物骨骼肌细胞的正常生肌分化和生脂转分化都具有重要的调节作用。     

单核苷酸多态性及其在猪育种中的应用

单核苷酸多态性是一类在基因组中最普遍存在的分子标记。本文综述了单核苷酸多态性的概念、检测技术及猪生长性状、背膘厚和瘦肉率、肉质性状和抗病性状中单核苷酸多态性的应用,以及SNP的应用前景。     

单核苷酸多态性及其在猪育种中的应用

单核苷酸多态性是一类在基因组中最普遍存在的分子标记。本文综述了单核苷酸多态性的概念、特点、检测技术及猪生长性状、背膘厚和瘦肉率、肉质性状和抗病性状中单梭苷酸多态性的应用。     

微卫星多态性与动物杂种优势预测

本文在动物杂种优势理论和对微卫星多态性产生机制、研究方法及分析技术的基础上,对微卫星多态性与动物杂种优势预测的关系及研究进展进行了综述,并对存在的问题和发展前景进行了探讨。     

MYOD基因家族在鸭小肠组织发育中的表达研究

为研究生肌调节因子MYOD家族对鸭小肠发育的影响,利用QRT-PCR法检测了MYOD家族各成员mRNA在鸭胚第10、14、18、22、27天及出生后1周小肠平滑肌发育过程中的相对表达量。结果表明,4个基因在鸭小肠发育过程中均有表达,且各自的表达量呈现一定的规律性变化。其中MYF5在第10天的表达量最高,之后总体呈下降趋势,到第27天时又达到一个高峰;MYOD1在第10天时表达量最高,之后急剧下降;MYOG的表达量从第10天开始曲折上升,到第22天时达到高峰,之后逐步下降;MYF6在第10天时表达量最高,之后急剧下降。初步推测,MYOD基因家族可能参与了小肠的发育或功能的调控。     

微卫星多态性与动物杂种优势预测

本文在动物杂种优势理论和对微卫星多态性产生机制、研究方法及分析技术的基础上，对微卫星多态性与动物杂种优势预测的关系及研究进展进行了综述，并对存在的问题和发展前景进行了探讨。     

猪肌细胞生成素基因的研究进展

综述猪肌细胞生成素(MyoG)对猪产肉量影响的作用机制和猪肌细胞生成素基因的定位及结构,并分析其PCR-RFLP遗传变异.     

北京鸭Myogenin基因部分序列的克隆及表达时间分析

利用4对引物分别对42、14日龄北京鸭胸肌组织RNA及出生后0日龄北京鸭腿肌组织和孵化22、15日龄胚胎腿肌RNA进行Myogenin基因的PCR反转录扩增，均没扩增出目的基因。资料分析表明在胚胎发育期内Myogenin基因可能只在成肌细胞分化的特定时间内表达，Myogenin基因也可在动物失去神经后或在体外培养的肌卫星细胞内表达。利用DNA扩增出了北京鸭Myogenin基因外显子1260bp部分序列，共编码86个氨基酸，编码氨基酸序列含有bHLH结构域，该结构与鸡、火鸡的同源性非常高。研究结果将为北京鸭Myogenin的表达、全序列的克隆及其分子标记的研究提供理论基础。     

Relation of porcine myogenin gene PCR/RFLP MspI and reproduction traits of the Czech Large White sows

Reproduction traits are highly important for pigs producers because of effect on economic efficiency. Hence, breeders as well as geneticists try to find the way to improve the reproduction traits. Because the protein coded by the myogenin gene (MYOG) is necessary for regulation of skeletal muscles development during embryogenesis, many authors have studied its influence on the meat traits of pigs. The aim of our study was to determine the effect of myogenin gene on the sows' reproduction traits. There were included 529 litters of 107 Czech Large White sows. Effects on the age of the first conception, service period, insemination index, average birth weight of piglets, average weights of litter at the age of 21 days, total number of born piglets, number of piglets born alive and number of weaned piglets were studied. For studying the influence of myogenin gene on chosen traits we used the mixed linear model procedure REML and in one case general linear model. Significant effects of myogenin gene on the age of first conception, the insemination index and on the litter size were proved.     

Influences of myogenin genotypes on birth weight, growth rate, carcass weight, backfat thickness, and lean weight of pigs.

Lean weight is related to muscle fiber number. Muscle fiber formation (myogenesis) occurs only during embryonic development when it is under the control of the MyoD gene family consisting of myogenin, MyoD1, myf-5, and myf-6. Myogenin has a central position within the MyoD gene family because myogenin expression abrogates myoblast proliferation potential and regulates the differentiation of single nucleated myoblasts into multinucleated myofibers. Thus, myogenin genotype could be related to variation in the number of muscle fibers formed, leading to variation in muscle mass and, thus, lean weight. A polymorphism at the porcine myogenin locus was associated with birth weight, growth rate, lean weight at 200 d, and backfat thickness. Yorkshire pigs from two commercial lines were genotyped, and crosses between heterozygous pigs and heterozygous and homozygous pigs were made. Resulting litters were genotyped, and phenotypic data were collected. Significant differences were found between the two homozygous myogenin genotypes for birth weight, growth rate, and lean weight, but not for backfat thickness. Variation at the myogenin locus explained 4% of the total phenotypic variation in birth weight, growth rate, and carcass weight, and 5.8% of the total variation in lean weight. We conclude that myogenin genotype influences porcine growth rate and muscle mass.     

猪骨骼肌卫星细胞中miR-24功能的初步研究及靶基因分析

为了探讨miR-24对猪骨骼肌发育的影响,对体外培养1日龄纯种长白公猪骨骼肌卫星细胞分别转染miR-24过表达载体和干扰片段,发现过表达miR-24后,成肌分化基因MyoD、Myogenin和Myf5表达量显著升高,骨骼肌卫星细胞分化趋势增强;干扰miR-24后,Myogenin的表达量显著降低;同时,靶基因预测结果显示Myogenin可能是miR-24的靶基因。     

影响瘦肉率的主基因及其分子标记的研究进展

近年来，影响瘦肉率及其相关的分子标记研究有一个巨大的飞跃，除已有较深入研究的氟烷基因外，还有肥胖基因及其受 体基因，胰岛素样生长因子2基因，肌肉生长和抑制素基因及脂肪酸结合蛋白基因等，本文综述了上述后4种基因的研究现状与前景，同时还简述了其他一些基因如肌浆蛋白基因，垂体特殊转录因子基因，原癌基因，激素敏感脂肪酶基因等。     

Messenger ribonucleic acid expression of the MyoD gene family in muscle tissue at slaughter in relation to selection for porcine growth rate

Livestock meat production capacity is related to muscle fiber numbers and growth. Muscle fibers develop during early embryonic development from proliferating and differentiating myoblasts. Post-natal muscle growth requires satellite cell proliferation and differentiation. Myoblast and satellite cell proliferation and differentiation is regulated by the genes of the MyoD gene family (myogenin, myf-5, myf-6, and MyoD1). Our aim was to study the mRNA expression of these genes in postnatal muscle tissue in relation to porcine selection for growth rate or leanness. Five boars from a line selected for fast growth (F-line) and five boars from a line selected against backfat thickness (L-line) were slaughtered, and biopsies were taken from 12 muscles. Between-line effects, within-line effects in relation to the performance of the pigs, and muscle-specific effects were studied. Comparing the F-line with the L-line revealed significantly greater myogenin, myf-5, and MyoD1 mRNA expression in some muscles of the F-line. The expression of myf-6 showed a tendency for the opposite effect in some muscles. Muscles were ordered by their muscle-specific growth rate (b-value). Within-line evaluation of the data revealed a systematic muscle effect for the myf-6 expression level in the F-line because higher b-values correlated with increased myf-6 expression level. Backfat thickness was negatively related to myogenin expression in the F-line. A relationship was found between myogenin:MyoD1 mRNA expression ratio and meat color/muscle fiber type composition in the L-line. Furthermore, the myogenin:MyoD1 ratio was greater in muscles from F-line boars than in muscles from L-line boars, which relates to the difference between the lines in muscle fiber type. We conclude that the mRNA levels of the MyoD genes in porcine muscle tissue at slaughter showed different relationships to selection for growth rate when evaluated between selection lines and within selection lines.     

Restriction fragment length polymorphisms in myogenin and myf3 genes and their influence on lean meat content in pigs

A total of 120 pigs from farm I and 109 from farm II were examined for Dde I polymorphism in myf3 and for Msp I polymorphism in myogenin genes. Meatness, weight and ratio of ham and loin meat, and loin eye area of animals with different genotypes for myf3 and for myogenin were compared. In myf3 polymorphism pigs A/A from farm I displayed significantly lower values for these characteristics. In farm II there was a reverse tendency. Present results indicate that the Dde I polymorphic region in porcine myf3 gene does not influence functionality of this gene but it may point at another polymorphism located nearby and being of functional importance. Two regions of the porcine myogenin gene were analysed for Msp I polymorphism. The region encompassing coding sequences ( MYOG1 ) was polymorphic only in three Pietrain pigs (out of 150 individuals tested) making it useless for assessment of meat productivity in pigs. The 3' untranslated region was polymorphic and the frequency of 4.2 and 4.9 alleles varied among breeds. The 4.2 variant was predominant in Pietrain pigs. Statistical analysis indicated that meat, ham and loin meat percentages as well as loin eye area were significantly higher in 4.2/4.2 homozygotes. Further study will be undertaken to explain the relations between meat productivity and myf3 and MYOG2 alleles within breed.     

In ovo L‐arginine supplementation stimulates myoblast differentiation but negatively affects muscle development of broiler chicken after hatching

In this study, we tested the hypothesis that in ovo feeding (IOF) of L‐arginine (L‐Arg) enhances nitric oxide (NO) production, stimulates the process of myogenesis, and regulates post‐hatching muscle growth. Different doses of L‐Arg were injected into the amnion of chicken embryos at embryonic day (ED) 16. After hatching, the body weight of individual male chickens was recorded weekly for 3 weeks. During in vitro experiments, myoblasts of the pectoralis major (PM) were extracted at ED16 and were incubated in medium containing 0.01 mm L‐Arg, 0.05 mm L‐Arg, and (or) 0.05 mm L‐nitro‐arginine‐methyl‐ester (L‐NAME), an inhibitor of nitric oxide synthase (NOS). When 25 mg/kg L‐Arg/initial egg weight was injected, no difference was observed in body weight at hatch, but a significant decrease was found during the following 3 weeks compared to that of the non‐injected and saline‐injected control, and this also affected the growth of muscle mass. L‐NAME inhibited gene expression of myogenic differentiation antigen (MyoD), myogenin, NOS, and follistatin, decreased the cell viability, and increased myostatin (MSTN) gene expression. 0.05 mm L‐Arg stimulated myogenin gene expression but also depressed muscle cell viability. L‐NAME blocked the effect of 0.05 mm L‐Arg on myogenin mRNA levels when co‐incubated with 0.05 mm L‐Arg. L‐Arg treatments had no significant influence on NOS mRNA gene expression, but had inhibiting effect on follistatin gene expression, while L‐NAME treatments had effects on both. These results suggested that L‐Arg stimulated myoblast differentiation, but the limited number of myoblasts would form less myotubes and then less myofibers, while the latter limited the growth of muscle mass.     

湖羊MyoG基因CDS的克隆及其真核表达载体的构建

参阅GenBank发表的波尔山羊肌细胞生成素(MyoG)基因序列,设计3对具有互补末端的特异性引物,分别扩增出湖羊MyoG基因的3个外显子,利用重叠PCR技术,将此3个外显子连接,并构建成重组质粒pcDNA3.0-MyoG,转染NIH-3T3细胞,RT-PCR鉴定。结果表明:成功克隆了湖羊MyoG基因的CDS,在离体细胞中MyoG基因发生了转录,这为进一步研究MyoG基因的体内外表达及生物学的活性奠定了基础。