The cloning, characterization, and expression profiling of the LRP8 gene in duck ( Anas platyrhynchos )
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The cloning, characterization, and expression profiling of the LRP8 gene in duck (Anas platyrhynchos) Shenqiang Hu • Hehe Liu • Zhixiong Pan • Fang Ding • Jie Kou • Liang Li • Jiwen Wang
Received: 19 August 2012 / Accepted: 23 November 2012 / Published online: 7 December 2012 Ó Springer Science+Business Media New York 2012
Abstract Low-density lipoprotein receptor-related protein 8 (LRP8) is a member of the low-density lipoprotein receptor gene family that functions in body lipoprotein homeostasis. In this study, reverse transcription-polymerase chain reaction, rapid amplification of cDNA ends, and real-time PCR were performed to characterize the duck LRP8 gene. The cDNA of duck LRP8 contained a 14-bp 50 UTR, a 2754-bp open reading frame, and a 189-bp 30 UTR. The duck LRP8 encoded a protein of 917 amino acid residues composed of five functional domains and resembling other members of the LDLR family, and it displayed high nucleotide and amino acid homology to the LRP8 sequences in other avian species. The mRNA expression level of LRP8 was greater in duck extra-hepatic adipose tissue than in the liver. The peak expression values of LRP8 in both liver and adipose tissues occurred at week 1 and were significantly higher than the values observed during any other week (p \ 0.05). Differences in the expression patterns of LRP8 mRNA from weeks 2 to 8 of growth were observed in different organs. A consistent low expression was observed in the liver, and fluctuating expression was observed in the subcutaneous adipose tissue (up- and then down-regulated) and abdominal adipose tissue (down-, then up-, then down-regulated). These findings suggest that LRP8 might play more important roles in regulating lipid metabolism in extra-hepatic adipose tissues than in the liver during early growth after hatching in the duck.
S. Hu H. Liu Z. Pan F. Ding J. Kou L. Li J. Wang (&) Institute of Animal Genetics and Breeding, College of Animal Science and Technology, Sichuan Agricultural University, Ya’an 625014, Sichuan, People’s Republic of China e-mail: [email protected]
Keywords
Duck LRP8 RACE Real-time PCR
Abbreviations ApoE Apolipoprotein E CM Chylomicron remnants DNL De novo lipogenesis HDL High-density lipoprotein LDL Low-density lipoprotein LDLR Low-density lipoprotein receptor LPL Lipoprotein lipase LRP8 Low-density lipoprotein receptor-related protein 8 TC Total cholesterol TG Triglycerides VLDL Very low-density lipoprotein
Introduction In avian species, unlike mammals, most of the endogenous body lipids originate from de novo lipogenesis in the liver, and lipogenesis in the adipose tissues is very limited [1]. The storage of lipids of hepatic origin in the body is dependent on the availability of the plasma lipoproteins involved in lipoprotein synthesis, secretion and transport [2]. The term ‘‘lipoproteins’’ refers to apolipoproteins, triglycerides (TG), cholesterol (TC), and phospholipids, which are grouped into different classes based on the proportions of lipids and proteins that they contain. The very lo
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