PLIN2基因在秦川肉牛皮下脂肪组织中的表达规律及基因多态性与肉质性状的关联分析

doi:10.16431/j.cnki.1671-7236.2018.06.020

摘要/Abstract

摘要：

试验旨在研究围脂滴蛋白2（PLIN2）基因在秦川肉牛皮下脂肪组织中的表达情况及基因多态性与肉质性状的关联分析。采用实时荧光定量PCR检测PLIN2基因mRNA在不同月龄（6~60月龄）秦川肉牛皮下脂肪组织中的表达情况；选取18~24月龄的健康秦川肉牛536头，采集血样并测定肉质性状指标，研究PLIN2基因单核苷酸多态性（SNP）和氨基酸突变，并与秦川肉牛肉质性状进行关联分析。结果发现，PLIN2基因在秦川肉牛皮下脂肪组织中的表达量呈先上升后下降趋势，18月龄时表达量最高。测序共发现9个SNPs位点，其中g.C7919>T位点不同基因型肌内脂肪含量差异显著（P<0.05），CT基因型显著高于TT和CC基因型；g.C7933>T位点不同基因型背膘厚及肌内脂肪含量均呈显著差异（P<0.05），CC和CT基因型个体背膘厚显著高于TT基因型，CT基因型个体肌内脂肪含量显著高于TT和CC基因型；g.G8015>C位点CC基因型个体肌内脂肪含量显著高于GG和GC基因型（P<0.05）；3'UTR区域g.T8496>C位点TC基因型个体肌内脂肪含量显著高于CC和TT基因型（P<0.05）；g.C8578>T位点TT基因型个体背膘厚显著高于CT和CC基因型（P<0.05）。综上，PLIN2基因对秦川肉牛肉质性状发育有一定影响，可作为秦川肉牛现代分子选育的候选参考基因。

关键词: 秦川肉牛; PLIN2基因; 表达谱; 多态性; 关联分析

Abstract:

This study investigated the expression of PLIN2 gene in subcutaneous adipose tissue and analyzed the association between PLIN2 gene single nucleotide polymorphism (SNPs) and meat quality traits in Qinchuan cattle.The mRNA expression levels of PLIN2 gene in subcutaneous adipose tissue from 6 to 60 months old Qinchuan cattle was detected by Real-time quantitative PCR.536 healthy Qinchuan cattles aged from 18 to 24 months old were selected to collect blood samples and determine the meat quality traits.The expression of PLIN2 gene in subcutaneous adipose tissue increased from 6 months old and peaked at 18 months old with the decreasing till 60 months old.The results showed that 9 SNPs of PLIN2 gene were identified by PCR amplification and direct sequencing,the intramuscular fat content (IMF) of CT genotype in g.C7919>T site was significantly higher than that of TT and CC genotypes (P<0.05).The back fat thickness (BFT) of CC and CT genotypes in g.C7933>T site were significantly higher than that of TT genotype (P<0.05),the IMF of CT genotype was significantly higher than that of TT and CC genotypes (P<0.05).The IMF of CC genotype in g.G8015>C site was significantly higher than that of GG and GC genotypes (P<0.05).The IMF of TC genotype in g.T8496>C site was significantly higher than that of CC and TT genotypes (P<0.05),the BFT of TT genotype in g.C8578>T site was significantly higher than that of CC and CT genotypes (P<0.05).PLIN2 gene could be considered as a candidate reference for Qinchuan cattle meat traits researching,and provided a certain basis for further breeding.

Key words: Qinchuan cattle; PLIN2 gene; expression pattern; polymorphism; association analysis

中图分类号:

S813.3

李佩韦, 吴森, 王亚宁, 王洪宝, 昝林森. PLIN2基因在秦川肉牛皮下脂肪组织中的表达规律及基因多态性与肉质性状的关联分析[J]. 《中国畜牧兽医》, 2018, 45(6): 1580-1589.

LI Peiwei, WU Sen, WANG Yaning, WANG Hongbao, ZAN Linsen. Expression Patterns of PLIN2 Gene in Subcutaneous Adipose Tissue and Association Analysis of Single Nucleotide Polymorphisms with Meat Quality Traits in Qinchuan Cattle[J]. , 2018, 45(6): 1580-1589.

参考文献

[1] MURPHY D J.The biogenesis and functions of lipid bodies in animals,plants and microorganisms[J].Progress in Lipid Research,2001,40(5):325-438.
[2] DAHLHOFF M,CAMERA E,PICARDO M,et al.PLIN2,the major perilipin regulated during sebocyte differentiation,controls sebaceous lipid accumulation in vitro and sebaceous gland size in vivo[J].Biochimica et Biophyssica Acta,2013,1830(10):4642-4649.
[3] MASUDA Y,ITABE H,ODAKI M, et al.ADRP/adipophilin is degraded through the proteasome-dependent pathway during regression of lipid-storing cells[J].Journal of Lipid Research,2006,47(1):87-98.
[4] MARTIN P,CEBO C,MIRANDA G.Interspecies Comparison of Milk Proteins:Quantitative Variability and Molecular Diversity[M].4th Edition.Advanced Dairy Chemistry:Proteins:Basic Aspects,2013.
[5] BICKEL E P,TANSEY T J,MICHAEL A W.PAT proteins,an ancient family of lipid droplet proteins that regulate cellular lipid stores[J].Biochimica et Biophysica Acta,2009,1791(6):419-440.
[6] LORD C C,RERGUSON D,THOMAS G,et al.Regulation of hepatic triacylglycerol metabolism by CGI-58 does not require ATGL co-activation[J].Cell Report,2016,16(4):939-949.
[7] DARWICH A A,PERREAU C,TSIKIS G,et al.Effect of different culture systems on adipocyte differentiation-related protein (ADRP) in bovine embryos[J].Animal Reprod Science,2014,145(3-4):105-113.
[8] WANG S,YANG H,WU J,et al.Metabolism as a tool for understanding human brain evolution:Lipid energy metabolism as an example[J].Journal of Human Evolution,2014,77(6):41-49.
[9] PROKESCH A,SMORLESI A,PERUGINI J,et al.Molecular aspects of adipoepithelial transdifferentiation in mouse mammary gland[J].Stem Cells,2014,32(10):2756-2766.
[10] WANG S,HWANG R D,GREENBERG S A,et al.Temporal and spatial assembly of lipid droplet-associated proteins in 3T3-L1 preadipocytes[J].Histochemistry and Cell Biology,2003,120(4):285-292.
[11] 李娜,李红波,周振勇,等.基因多态性与超声波活体测定相结合对肉牛主要肉质性状的研究[J].中国畜牧兽医,2017,44(3):825-831. LI N,LI H B,ZHOU Z Y,et al.Study on genetic polymorphism and ultrasound measurements for major meat quality traits in beef cattle[J].China Animal Husbandry & Veterinary Medicine,2017,44(3):825-831.(in Chinese)
[12] GUI L,HAO R,ZHANG Y,et al.Haplotype distribution in the class Ⅰ sirtuin genes and their associations with ultrasound carcass traits in Qinchuan cattle (Bos taurus)[J].Molecular and Cellular Probes,2015,29(3):167-171.
[13] BOSMA M,RONNIE M,LAUREN M S,et al.The lipid droplet coat protein perilipin 5 also localizes to muscle mitochondria[J].Histochemistry and Cell Biology,2012,137(2):205-216.
[14] RODEHEFFER S M,BIRSOY K,FRIEDMAN M J.Identification of white adipocyte progenitor cells in vivo[J].Cell, 2008,135(2):240-249.
[15] MÉLANLE F.Human hormone-sensitive lipase (HSL):Expression in white fat corrects the white adipose phenotype of HSL-deficient mice[J].The Journal of Lipid Research,2005,46(9):1860-1867.
[16] ESCRIBA V P,BUSQUETS X,INOKUCHI J,et al.Membrane lipid therapy:Modulation of the cell membrane composition and structure as a molecular base for drug discovery and new disease treatment[J].Progress in Lipid Research,2015,59(4):38-53.
[17] SASTRE D,NATHÁLIA N C,ANDRÉ L S,et al.Expression of PLIN2 and PLIN3 during oocyte maturation and early embryo development in cattle[J].Theriogenology,2014,81(2):326-331.
[18] 昝林森,莫放,周贵,等.牛生产学[M].第2版.北京:中国农业出版社,2007. ZAN L S,MO F,ZHOU G,et al.Cattle Produc-tion[M].2nd Edition.Beijing:China Agriculture Press,2007.(in Chinese)
[19] 胡鑫,王国富,吴慧光,等.中国西门塔尔牛血管生成素样蛋白3基因多态性及其与肉质性状的关联性分[J].中国畜牧兽医,2015,42(2):438-442. HU X,WANG G F,WU H G,et al.Polymorphism of ANGPTL3 gene and its association analysis with meat traits in Chinese simmental cattle[J].China Animal Husbandry & Veterinary Medicine,2015,42(2):438-442.(in Chinese)
[20] MAK K M,REN C,PONOMARENKO A,et al.Adipose differentiation-related protein is a reliable lipid droplet marker in alcoholic fatty liver of rats[J].Alcoholism:Clinical Experimental Research,2008,32(4):683-689.
[21] DAVOLI R,GANDOLFI G,BRAGLIA S,et al.New SNP of the porcine perilipin 2(PLIN2) gene,association with carcass traits and expression analysis in skeletal muscle[J].Molecular Biology Reports, 2011,38(3):1575-1583.
[22] MCMANAMAN J L,ELISE S B,DAVID J O,et al.Perilipin-2-null mice are protected against diet-induced obesity,adipose inflammation,and fatty liver disease[J].Journal of Lipid Research,2013,54(5):1346-1359.
[23] LEFEVR C,JOBARD F,BOUADJAR B,et al.Mutations in CGI-58,the gene encoding a new protein of the esterase/lipase/thioesterase subfamily,in Chanarin-Dorfman syndrome[J].American Journal of Human Genetics, 2001,69(5):1002-1012.
[24] LASS A,ZIMMERMANN R,HAEMMERLE G,et al.Adipose triglyceride lipase-mediated lipolysis of cellular fat stores is activated by CGI-58 and defective in Chanarin-Dorfman syndrome[J].Cell Metabolism, 2006,3(5):309-319.