Sun Y N, Gao Y, Qiao S P, Wang S Z, Duan K, Wang Y X, Li H, Wang N
Key Laboratory of Chicken Genetics and Breeding, Ministry of Agriculture; Key Laboratory of Animal Genetics, Breeding and Reproduction, Education Department of Heilongjiang Province; College of Animal Science and Technology, Northeast Agricultural University, Harbin, 150030, P. R. China.
J Anim Sci. 2014 Jan;92(1):48-53. doi: 10.2527/jas.2013-6962. Epub 2013 Dec 4.
Peroxisome proliferator-activated receptor γ is a master regulator of adipocyte differentiation and function. Expression of PPARγ in mammals is regulated by DNA methylation; however, it is currently unknown whether chicken PPARγ expression is regulated by DNA methylation. To enhance our understanding of molecular mechanisms underlying chicken adipose tissue development and adipogenesis, we investigated the promoter methylation status and gene expression of PPARγ gene in Northeast Agricultural University broiler lines divergently selected for abdominal fat content (NEAUHLF). Deoxyribonucleic acid methylation was analyzed by bisulfite sequencing method, and mRNA expression was detected by real-time quantitative real time reverse-transcription polymerase chain reaction (RT-PCR). The analyzed region located from -1,175 to -301 bp upstream of the translation start codon ATG contains 6 CpG dinucleotides, which are located at positions -1,014, -796, -625, -548, -435, and -383 bp, respectively. The results revealed that the 3 CpGs at positions -548, -435, and -383 bp showed differential methylation between the lean and fat chicken lines, but the other 3 CpG sites at positions -1,014, -796, and -625 bp did not. PPARγ gene promoter methylation in both chicken lines decreased with age, and PPARγ promoter methylation levels were significantly higher in lean than fat broilers at 2 wk of age (79.9 to 64.5%; P < 0.0001), at 3 wk of age (66.7 to 58.3%; P < 0.0001), and at 7 wk of age (50.0 to 42.7%; P = 0.0004). Real-time quantitative RT-PCR analysis showed that, negatively correlated with DNA methylation (Pearson's r = -0.653, P = 0.0057), PPARγ expression was increased with age and significantly lower in lean than fat chicken lines at 2, 3, and 7 wk of age (P < 0.0001). In conclusion, our findings suggest that chicken PPARγ is regulated by DNA methylation during adipose tissue development.
过氧化物酶体增殖物激活受体γ是脂肪细胞分化和功能的主要调节因子。哺乳动物中PPARγ的表达受DNA甲基化调控;然而,目前尚不清楚鸡PPARγ的表达是否受DNA甲基化调控。为了加深我们对鸡脂肪组织发育和脂肪生成潜在分子机制的理解,我们研究了东北农业大学高、低脂双向选择肉鸡品系(NEAUHLF)中PPARγ基因的启动子甲基化状态和基因表达。采用亚硫酸氢盐测序法分析DNA甲基化,通过实时定量逆转录聚合酶链反应(RT-PCR)检测mRNA表达。分析区域位于翻译起始密码子ATG上游-1175至-301 bp,包含6个CpG二核苷酸,分别位于-1014、-796、-625、-548、-435和-383 bp位置。结果显示,-548、-435和-383 bp位置的3个CpG在低脂和高脂鸡品系间表现出差异甲基化,而-1014、-796和-625 bp位置的其他3个CpG位点则没有。两个鸡品系中PPARγ基因启动子甲基化均随年龄增长而降低,在2周龄(79.9%至64.5%;P<0.0001)、3周龄(66.7%至58.3%;P<0.0001)和7周龄(50.0%至42.7%;P = 0.0004)时,低脂肉鸡的PPARγ启动子甲基化水平显著高于高脂肉鸡。实时定量RT-PCR分析表明,PPARγ表达与DNA甲基化呈负相关(Pearson's r = -0.653,P = 0.0057),随年龄增长而增加,在2、3和7周龄时,低脂鸡品系中的PPARγ表达显著低于高脂鸡品系(P<0.0001)。总之,我们的研究结果表明,鸡PPARγ在脂肪组织发育过程中受DNA甲基化调控。