SYBR Green Ⅰ荧光定量PCR法检测朗德鹅填饲前后黑素皮质素受体-4基因(MC4R)的表达  

The Melanocortin-4 Receptor Gene(MC4R) Expression Detected By SYBR Green Ⅰ Real-time Quantitative PCR in Landes Geese before and after Overfeeding

作　　者：原昊[1,2] 张红艳[1,3] 田勇[1] 沈军达[1] 王芳[1,4] 弓彦[1,2] 王德前[1] 石放雄[2] 卢立志[1] 

机构地区：[1]浙江省农业科学院畜牧兽医研究所,杭州310021 [2]南京农业大学动物科技学院,南京210095 [3]温州医学院,温州325000 [4]浙江大学动物科学学院,杭州310020

出　　处：《农业生物技术学报》2011年第4期692-697,共6页Journal of Agricultural Biotechnology

基　　金：国家重点国际合作项目(No.2008DFA31060);浙江省重大科技攻关项目(No.2007C12058)共同资助

摘　　要：黑素皮质素受体4基因(MC4R)在各组织中的不同表达对于鹅脂肪代谢具有重要作用。本研究应用强制填饲技术建立腹型肥胖朗德鹅(Casuarius casuarius)模型,提取填饲后及正常朗德鹅新鲜腹脂、肝脏等13种组织的总RNA,SYBRGreenⅠ荧光定量PCR法检测各组织MC4R基因的表达,选择β-actin作为内参基因,使用半定量2-ΔΔCt法分析。PCR结果显示,MC4R基因及β-actin在填饲前后朗德鹅心脏、肝脏、脾脏、肺、肾脏、肌胃、小肠、胰腺、脑、胸肌、腿肌、腹脂、皮下脂等13种组织中均存在不同程度表达。SYBRGreenⅠ荧光定量PCR结果表明,在肾、小肠和心这3个组织中,填饲后朗德鹅MC4R表达量均比对照组中朗德鹅该基因表达量低,填饲后分别为填饲前表达量的0.41±1.80、0.65±5.75和0.72±1.22倍。而在其他组织中,MC4R基因表达量均有不同程度的增加,填饲组/对照组相对表达量倍数由高到低顺序分别为:22.78±1.00倍(脑),9.08±2.80倍(肝),5.28±1.83倍(肺),3.78±3.01倍(胰),3.07±3.64倍(腿肌),2.90±0.97倍(胃),2.34±1.66倍(皮下脂肪),2.18±3.01倍(腹脂),2.07±0.37倍(胸肌),2.01±1.75倍(脾)。结果符合MC4R作用机理,为畜牧生产及人类肥胖问题的研究提供间接依据。The expression characterization of Melanocortin-4 Receptor Gene（MC4R） before and after overfeeding plays an important role in fat metabolism of Lades geese（Casuarius casuarius）.SYBR GreenⅠReal-time quantitative PCR was developed to analyze the expression characterization of MC4R gene in 13 tissues.PCR results showed that both MC4R and β-actin were expressed in all 13 tissues before and after overfeeding of Landes goose.SYBR GreenⅠReal-time quantitative PCR results showed that the relative expression of MC4R in kidney,intestine and heart after overfeeding was lower than those before overfeeding,their 2^-ΔΔCt values were 0.41±1.80,0.65±5.75 and 0.72±1.22,respectively.But the relative expressions of MC4R in other tissues were all higher after overfeeding,their 2-ΔΔCt values from high to low were 22.78±1.00（brain）,9.08±2.80（liver）,5.28±1.83（lung）,3.78±3.01（pancreas）,3.07±3.64（leg muscle）,2.90±0.97（stomach）,2.34±1.66（subcutaneous adipose）,2.18±3.01（abdominal adipose）,2.07±0.37（chest muscle） and 2.01±1.75（pleen）.We conclude that the results are consistented with the mechanism of MC4R and can provide indirect basis for researches on animal production and human obesity.

关 键 词：黑素皮质素受体4基因(MC4R) 脂肪代谢 鹅 实时荧光定量PCR 表达 

分 类 号：S831.2[农业科学—畜牧学]