Molecular and Cellular Biochemistry

, Volume 375, Issue 1–2, pp 139–149 | Cite as

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
Article
First Online:
Received:
Accepted:
  • 348 Downloads

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 5′ UTR, a 2754-bp open reading frame, and a 189-bp 3′ 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.

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

This is a preview of subscription content, log in to check access.

Notes

Acknowledgments

The study was supported by the earmarked fund for National High Technology Research and Development Program of China (No. 2010AA10A109), the National Waterfowl Industrial Technology System (No. CARS-43-6), and the Program for Technology Innovative Research Team of Sichuan Province of China (No. 2011JTD0032).

References

  1. 1.
    Saadoun A, Leclercq B (1987) In vivo lipogenesis of genetically lean and fat chickens: effects of nutritional state and dietary fat. J Nutr 117(3):428–435PubMedGoogle Scholar
  2. 2.
    Hermier D (1997) Lipoprotein metabolism and fattening in poultry. J Nutr 127(5 Suppl):805S–808SPubMedGoogle Scholar
  3. 3.
    Cryer A (1981) Tissue lipoprotein lipase activity and its action in lipoprotein metabolism. Int J Biochem 13(5):525–541PubMedCrossRefGoogle Scholar
  4. 4.
    Goldstein JL, Brown MS, Anderson RG, Russell DW, Schneider WJ (1985) Receptor-mediated endocytosis: concepts emerging from the LDL receptor system. Annu Rev Cell Biol 1:1–39. doi: 10.1146/annurev.cb.01.110185.000245 PubMedCrossRefGoogle Scholar
  5. 5.
    Kim DH, Iijima H, Goto K, Sakai J, Ishii H, Kim HJ, Suzuki H, Kondo H, Saeki S, Yamamoto T (1996) Human apolipoprotein E receptor 2. A novel lipoprotein receptor of the low density lipoprotein receptor family predominantly expressed in brain. J Biol Chem 271(14):8373–8380PubMedCrossRefGoogle Scholar
  6. 6.
    Willnow TE (1999) The low-density lipoprotein receptor gene family: multiple roles in lipid metabolism. J Mol Med (Berl) 77(3):306–315CrossRefGoogle Scholar
  7. 7.
    Mahley RW (1988) Apolipoprotein E: cholesterol transport protein with expanding role in cell biology. Science 240(4852):622–630PubMedCrossRefGoogle Scholar
  8. 8.
    Greenow K, Pearce NJ, Ramji DP (2005) The key role of apolipoprotein E in atherosclerosis. J Mol Med (Berl) 83(5):329–342. doi: 10.1007/s00109-004-0631-3 CrossRefGoogle Scholar
  9. 9.
    Shen GQ, Li L, Girelli D, Seidelmann SB, Rao S, Fan C, Park JE, Xi Q, Li J, Hu Y, Olivieri O, Marchant K, Barnard J, Corrocher R, Elston R, Cassano J, Henderson S, Hazen SL, Plow EF, Topol EJ, Wang QK (2007) An LRP8 variant is associated with familial and premature coronary artery disease and myocardial infarction. Am J Hum Genet 81(4):780–791. doi: S0002-9297(07)63053-5 PubMedCrossRefGoogle Scholar
  10. 10.
    Martinelli N, Olivieri O, Shen GQ, Trabetti E, Pizzolo F, Busti F, Friso S, Bassi A, Li L, Hu Y, Pignatti PF, Corrocher R, Wang QK, Girelli D (2009) Additive effect of LRP8/APOER2 R952Q variant to APOE epsilon2/epsilon3/epsilon4 genotype in modulating apolipoprotein E concentration and the risk of myocardial infarction: a case-control study. BMC Med Genet 10:41. doi: 1471-2350-10-41 PubMedCrossRefGoogle Scholar
  11. 11.
    Dolley G, Lamarche B, Despres JP, Bouchard C, Perusse L, Vohl MC (2011) Investigation of LRP8 gene in 1p31 QTL linked to LDL peak particle diameter in the Quebec family study. Mol Genet Metab 102(4):448–452. doi: S1096-7192(10)00586-X PubMedCrossRefGoogle Scholar
  12. 12.
    Wang L, Wang X, Laird N, Zuckerman B, Stubblefield P, Xu X (2006) Polymorphism in maternal LRP8 gene is associated with fetal growth. Am J Hum Genet 78(5):770–777. doi: S0002-9297(07)63812-9 PubMedCrossRefGoogle Scholar
  13. 13.
    Duit S, Mayer H, Blake SM, Schneider WJ, Nimpf J (2010) Differential functions of ApoER2 and very low density lipoprotein receptor in Reelin signaling depend on differential sorting of the receptors. J Biol Chem 285(7):4896–4908. doi: M109.025973 PubMedCrossRefGoogle Scholar
  14. 14.
    Argov N, Moallem U, Sklan D (2004) Lipid transport in the developing bovine follicle: messenger RNA expression increases for selective uptake receptors and decreases for endocytosis receptors. Biol Reprod 71(2):479–485. doi: 10.1095/biolreprod.104.028092 PubMedCrossRefGoogle Scholar
  15. 15.
    Trommsdorff M, Gotthardt M, Hiesberger T, Shelton J, Stockinger W, Nimpf J, Hammer RE, Richardson JA, Herz J (1999) Reeler/disabled-like disruption of neuronal migration in knockout mice lacking the VLDL receptor and ApoE receptor 2. Cell 97(6):689–701. doi: S0092-8674(00)80782-5 PubMedCrossRefGoogle Scholar
  16. 16.
    de Groot PG, van Lummel M, Pennings M, Urbanus R, Bas de Laat H, Lenting PJ, Derksen RH (2004) Beta2-glycoprotein I and LDL-receptor family members. Thromb Res 114(5–6):455–459. doi: S0049384804003226 PubMedCrossRefGoogle Scholar
  17. 17.
    Jeon H, Blacklow SC (2005) Structure and physiologic function of the low-density lipoprotein receptor. Annu Rev Biochem 74:535–562. doi: 10.1146/annurev.biochem.74.082803.133354 PubMedCrossRefGoogle Scholar
  18. 18.
    Russell DW, Brown MS, Goldstein JL (1989) Different combinations of cysteine-rich repeats mediate binding of low density lipoprotein receptor to two different proteins. J Biol Chem 264(36):21682–21688PubMedGoogle Scholar
  19. 19.
    Huang W, Dolmer K, Gettins PG (1999) NMR solution structure of complement-like repeat CR8 from the low density lipoprotein receptor-related protein. J Biol Chem 274(20):14130–14136PubMedCrossRefGoogle Scholar
  20. 20.
    Davis CG, Goldstein JL, Sudhof TC, Anderson RG, Russell DW, Brown MS (1987) Acid-dependent ligand dissociation and recycling of LDL receptor mediated by growth factor homology region. Nature 326(6115):760–765. doi: 10.1038/326760a0 PubMedCrossRefGoogle Scholar
  21. 21.
    May P, Bock HH, Nimpf J, Herz J (2003) Differential glycosylation regulates processing of lipoprotein receptors by gamma-secretase. J Biol Chem 278(39):37386–37392. doi: 10.1074/jbc.M305858200 PubMedCrossRefGoogle Scholar
  22. 22.
    Iijima H, Miyazawa M, Sakai J, Magoori K, Ito MR, Suzuki H, Nose M, Kawarabayasi Y, Yamamoto TT (1998) Expression and characterization of a very low density lipoprotein receptor variant lacking the O-linked sugar region generated by alternative splicing. J Biochem 124(4):747–755PubMedCrossRefGoogle Scholar
  23. 23.
    Chen WJ, Goldstein JL, Brown MS (1990) NPXY, a sequence often found in cytoplasmic tails, is required for coated pit-mediated internalization of the low density lipoprotein receptor. J Biol Chem 265(6):3116–3123PubMedGoogle Scholar
  24. 24.
    Korschineck I, Ziegler S, Breuss J, Lang I, Lorenz M, Kaun C, Ambros PF, Binder BR (2001) Identification of a novel exon in apolipoprotein E receptor 2 leading to alternatively spliced mRNAs found in cells of the vascular wall but not in neuronal tissue. J Biol Chem 276(16):13192–13197. doi: 10.1074/jbc.M011795200 PubMedCrossRefGoogle Scholar
  25. 25.
    Riddell DR, Vinogradov DV, Stannard AK, Chadwick N, Owen JS (1999) Identification and characterization of LRP8 (apoER2) in human blood platelets. J Lipid Res 40(10):1925–1930PubMedGoogle Scholar
  26. 26.
    Pennings MT, Derksen RH, Urbanus RT, Tekelenburg WL, Hemrika W, de Groot PG (2007) Platelets express three different splice variants of ApoER2 that are all involved in signaling. J Thromb Haemost 5(7):1538–1544. doi: JTH02605 PubMedCrossRefGoogle Scholar
  27. 27.
    Novak S, Hiesberger T, Schneider WJ, Nimpf J (1996) A new low density lipoprotein receptor homologue with 8 ligand binding repeats in brain of chicken and mouse. J Biol Chem 271(20):11732–11736PubMedCrossRefGoogle Scholar
  28. 28.
    Perez-Garcia CG, Tissir F, Goffinet AM, Meyer G (2004) Reelin receptors in developing laminated brain structures of mouse and human. Eur J Neurosci 20(10):2827–2832. doi: EJN3733 PubMedCrossRefGoogle Scholar
  29. 29.
    Lambson RO (1970) An electron microscopic study of the entodermal cells of the yolk sac of the chick during incubation and after hatching. Am J Anat 129(1):1–19. doi: 10.1002/aja.1001290102 PubMedCrossRefGoogle Scholar
  30. 30.
    O’Hea EK, Leveille GA (1969) Lipid biosynthesis and transport in the domestic chick (Gallus domesticus). Comp Biochem Physiol 30(1):149–159PubMedCrossRefGoogle Scholar
  31. 31.
    Foglia TA, Cartwright AL, Gyurik RJ, Philips JG (1994) Fatty acid turnover rates in the adipose tissues of the growing chicken (Gallus domesticus). Lipids 29(7):497–502PubMedCrossRefGoogle Scholar
  32. 32.
    Ding F, Pan Z, Kou J, Li L, Xia L, Hu S, Liu H, Wang J (2012) De novo lipogenesis in the liver and adipose tissues of ducks during early growth stages after hatching. Comp Biochem Physiol B 163(1):154–160. doi: S1096-4959(12)00099-1 PubMedCrossRefGoogle Scholar
  33. 33.
    Whitehead CC, Griffin HD (1984) Development of divergent lines of lean and fat broilers using plasma very low density lipoprotein concentration as selection criterion: the first three generations. Br Poult Sci 25(4):573–582. doi: 10.1080/00071668408454899 PubMedCrossRefGoogle Scholar
  34. 34.
    Reddy SS, Connor TE, Weeber EJ, Rebeck W (2011) Similarities and differences in structure, expression, and functions of VLDLR and ApoER2. Mol Neurodegener 6:30. doi: 1750-1326-6-30 PubMedCrossRefGoogle Scholar
  35. 35.
    Ndiaye K, Fayad T, Silversides DW, Sirois J, Lussier JG (2005) Identification of downregulated messenger RNAs in bovine granulosa cells of dominant follicles following stimulation with human chorionic gonadotropin. Biol Reprod 73(2):324–333PubMedCrossRefGoogle Scholar
  36. 36.
    Andersen OM, Yeung CH, Vorum H, Wellner M, Andreassen TK, Erdmann B, Mueller EC, Herz J, Otto A, Cooper TG, Willnow TE (2003) Essential role of the apolipoprotein E receptor-2 in sperm development. J Biol Chem 278(26):23989–23995. doi: 10.1074/jbc.M302157200 PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2012

Authors and Affiliations

  • Shenqiang Hu
    • 1
  • Hehe Liu
    • 1
  • Zhixiong Pan
    • 1
  • Fang Ding
    • 1
  • Jie Kou
    • 1
  • Liang Li
    • 1
  • Jiwen Wang
    • 1
  1. 1.Institute of Animal Genetics and Breeding, College of Animal Science and TechnologySichuan Agricultural UniversityYa’anPeople’s Republic of China

Personalised recommendations