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Gene expression profiles analyzed by DNA sequencing of cDNA clones constructed from porcine preadipocytes and adipocytes

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Abstract

As a step towards understanding the molecular mechanism of adipogenesis in pigs, preadipocytes purified from the back fat of 1 day-old female piglets were used for in vitro culture. Normalized cDNA libraries were constructed with 1.6×107 and 1.1×107 independent clones from preadipocyte and mature adipocyte mRNAs, respectively. Polymerase chain reaction (PCR) result using primers T3 and T7 (universal primer) confirmed the presence of the insert in the vector. Sequencing of 2,112 randomly selected clones from each cDNA library identified 217 clusters, 1,169 singletons, and 216 contigs in preadipocytes and 231 clusters, 1,100 singletons, and 233 contigs in mature adipocytes. Expressed sequence tag (EST) identified 24 genes with known annotation highly expressed in adipocytes and 21 in preadipocytes by at least four EST number. Among those 45 genes, when analyzed by real time RT-PCR, 76% of the gene showed significant difference between preadipocytes and mature adipocytes. Highly expressed genes in mature adipocytes were related to adipogenesis, extracellular matrix control and oncogenes, whereas cytoskeleton-related genes were down-regulated. An interesting similarity found during gene profile studies indicated a correlation between cancer and adipogenesis.

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References

  • Birnboim HC and Doly J (1979) A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res. 7: 1513–1523.

    Article  PubMed  CAS  Google Scholar 

  • Bourlier V, Zakaroff-Girard A, De Barros S, Pizzacalla C, de Saint Front VD, Lafontan M, Bouloumié A, and Galitzky J (2005) Protease inhibitor treatments reveal specific involvement of matrix metalloproteinase-9 in human adipocyte differentiation. J. Pharmacol. Exp. Ther. 312: 1272–1279.

    Article  PubMed  CAS  Google Scholar 

  • Cannarile L, Zollo O, D’Adamio F, Ayroldi E, Marchetti C, Tabilio A, Bruscoli S, and Riccardi C (2001) Cloning, chromosomal assignment and tissue distribution of human GILZ, a glucocorticoid hormone-induced gene. Cell Death Differ. 8: 201–203.

    Article  PubMed  CAS  Google Scholar 

  • Chiellini C, Costa M, Novelli SE, Amri EZ, Benzi L, Bertacca A, Cohen P, Del Prato S, Friedman JM, and Maffei M (2003) Identification of cathepsin K as a novel marker of adiposity in white adipose tissue. J. Cell Physiol. 195: 309–321.

    Article  PubMed  CAS  Google Scholar 

  • Croissandeau G, Chrétien M, and Mbikay M (2002) Involvement of matrix metalloproteinases in the adipose conversion of 3T3-L1 preadipocytes. Biochem. J. 364: 739–746.

    Article  PubMed  CAS  Google Scholar 

  • Festa M, Ricciardelli G, Mele G, Pietropaolo C, Ruffo A, and Colonna A (2000) Overexpression of H ferritin and up-regulation of iron regulatory protein genes during differentiation of 3T3-L1 pre-adipocytes. J. Biol. Chem. 275: 36708–36712.

    Article  PubMed  CAS  Google Scholar 

  • Feve B (2005) Adipogenesis: cellular and molecular aspects. Best Pract. Res. Clin. Endocrinol. Metab. 19: 483–499.

    Article  PubMed  CAS  Google Scholar 

  • Flake GP, Andersen J, and Dixon D (2003) Etiology and pathogenesis of uterine leiomyoma. A review. Environ. Health. Perspect. 11: 1037–1054.

    Google Scholar 

  • Frlan R and Gobec S (2006) Inhibitors of cathepsin B. Curr. Med. Chem. 13: 2309–2327.

    Article  PubMed  CAS  Google Scholar 

  • Fu Y, Luo N, Klein RL, and Garvey WT (2005) Adiponectin promotes adipocyte differentiation, insulin sensitivity, and lipid accumulation. J. Lipid Res. 46: 1369–1379.

    Article  PubMed  CAS  Google Scholar 

  • Hajra AK, Larkins LK, Das AK, Hemati N, Erickson RL, and MacDougald OA (2000) Induction of the peroxisomal glycerolipid-synthesizing enzymes during differentiation of 3T3-L1 adipocytes. Role in triacylglycerol synthesis. J. Biol. Chem. 275: 9441–9446.

    Article  PubMed  CAS  Google Scholar 

  • Holst D and Grimaldi PA (2002) New factors in the regulation of adipocyte differentiation and metabolism. Curr. Opin. Lipidol. 13: 241–245.

    Article  PubMed  CAS  Google Scholar 

  • Jongeneel CV (2000) Searching the expressed sequence tag (EST) databases: panning for genes. Brief. Bioinform. 1: 76–92.

    Article  PubMed  CAS  Google Scholar 

  • Kaartinen MT, Murshed M, Karsenty G, and McKee MD (2007) Osteopontin upregulation and polymerization by transglutaminase 2 in calcified arteries of Matrix Gla protein-deficient mice. J. Histochem. Cytochem. 55: 375–386.

    Article  PubMed  CAS  Google Scholar 

  • Karbowska J and Kochan Z (2006) Role of adiponectin in the regulation of carbohydrate and lipid metabolism. J. Physiol. Pharmacol. 57Suppl 6: 103–113.

    PubMed  Google Scholar 

  • Kasai H, Nadano D, Hidaka E, Higuchi K, Kawakubo M, Sato TA, and Nakayama J (2003) Differential expression of ribosomal proteins in human normal and neoplastic colorectum. J. Histochem. Cytochem. 51: 567–574.

    Article  PubMed  CAS  Google Scholar 

  • Kawaguchi N, Sundberg C, Kveiborg M, Moghadaszadeh B, Asmar M, Dietrich N, Thodeti CK, Nielsen FC, Moller P, Mercurio AM, et al. (2003) ADAM12 induces actin cytoskeleton and ex tracellular matrix reorganization during early adipocyte differentiation by regulating β1 intergrin function. J. Cell. Sci. 116: 3893–3904.

    Article  PubMed  CAS  Google Scholar 

  • Kelley JM, Field CE, Craven MB, Bocskai D, Kim UJ, Rounsley SD, and Adams MD (1999) High throughput direct end sequencing of BAC clones. Nucleic. Acids Res. 27: 1539–1546.

    Article  PubMed  CAS  Google Scholar 

  • Kim JB and Spiegelman BM (1996) ADD1/SREBP1 promotes adipocyte differentiation and gene expression linked to fatty acid metabolism. Genes Dev. 10: 1096–1107.

    Article  PubMed  CAS  Google Scholar 

  • Lee EJ, Bajracharya P, Jang EJ, Chang JS, Lee HJ, Hong SK, and Choi I (2010) Effect of sex steroid hormones on bovine myogenic satellite cell proliferation, differentiation and lipid accumulation in myotube. Asian-Aust. J. Anim. Sci. 23: 649–658.

    CAS  Google Scholar 

  • Miller RE, Hackenberg RW, and Gershman H (1978) Regulation of glutamine synthetase in cultured 3T3-L1 cells by insulin, hydrocortisone, and dibutyryl cyclic AMP. Proc. Natl. Acad. Sci. USA 75: 1418–1422.

    Article  PubMed  CAS  Google Scholar 

  • Moller DE and Flier JS (1991) Insulin resistance: mechanisms, syndromes, and implication. N. Engl. J. Med. 325: 938–948.

    Article  PubMed  CAS  Google Scholar 

  • Morita Y, Araki H, Sugimoto T, Takeuchi K, Yamane T, Maeda T, Yamamoto Y, Nishi K, Asano M, Shirahama-Noda K, et al. (2007) Legumain/asparaginyl endopeptidase controls extracellular matrix remodeling through the degradation of fibronectin in mouse renal proximal tubular cells. FEBS Lett. 581: 1417–1424.

    Article  PubMed  CAS  Google Scholar 

  • Murata Y, Sun-Wada GH, Yoshimizu T, Yamamoto A, Wada Y, and Futai M (2002) Differential localization of the vacuolar H+ pump with G subunit isoforms (G1 and G2) in mouse neurons. J. Biol. Chem. 277: 36296–36303.

    Article  PubMed  CAS  Google Scholar 

  • Mutch DM, Rouault C, Keophiphath M, Lacasa D, and Clément K (2009) Using gene expression to predict the secretome of differentiating human preadipocytes. Int. J. Obes. (Lond). 33: 354–363.

    Article  CAS  Google Scholar 

  • Nagaraj SH, Gasser RB, and Ranganathan S (2006) A hitchhiker’s guide to expressed sequence tag (EST) analysis. Brief. Bioinform. 8: 16–21.

    Article  Google Scholar 

  • Nilsson J, Koskiniemi S, Persson K, Grahn B, and Holm I (1997) Polyamines regulate both transcription and translation of the gene encoding ornithine decarboxylase antizyme in mouse. Eur. J. Biochem. 250: 223–231.

    Article  PubMed  CAS  Google Scholar 

  • Ntambi JM and Kim YC (2000) Adipocyte differentiation and gene expression. J. Nutr. 130: 3122S–3126S.

    PubMed  CAS  Google Scholar 

  • Pajak B and Orzechowski A (2006) Clusterin: the missing link in the calcium-dependent resistance of cancer cells to apoptogenic stimuli. Postepy Hig. Med. Dosw. 60: 45–51.

    Google Scholar 

  • Pang WJ, Yu TY, Bai L, Yang YJ, and Yang GS (2009) Tissue expression of porcine FoxO1 and its negative regulation during primary preadipocyte differentiation. Mol. Biol. Rep. 36: 165–176.

    Article  PubMed  CAS  Google Scholar 

  • Poirier P and Eckel RH (2002) Obesity and cardiovascular disease. Current Atherosclerosis Reports. 4: 448–453.

    Article  PubMed  Google Scholar 

  • Rizzi F, Belloni L, Crafa P, Lazzaretti M, Remondini D, Ferretti S, Cortellini P, Corti A, and Bettuzzi S (2008) A novel gene signature for molecular diagnosis of human prostate cancer by RT-qPCR. PLoS One 3: e3617.

    Article  PubMed  Google Scholar 

  • Saladin R, Fajas L, Dana S, Halvorsen YD, Auwerx J, and Briggs M (1999) Differential regulation of peroxisome proliferator activated receptor g1 (PPARg1) and PPARg2 mRNA expression in early stages of adipogenesis. Cell Growth Differ. 10: 43–48.

    PubMed  CAS  Google Scholar 

  • Samulin J, Berget I, Lien S, and Sundvold H (2008) Differential gene expression of fatty acid binding proteins during porcine adipogenesis. Comp. Biochem. Physiol. B. Biochem. Mol. Biol. 151: 147–152.

    Article  PubMed  Google Scholar 

  • Spiegelman BM, Choy L, Hotamisligil GS, Graves RA, and Tontonoz P (1993) Regulation of adipocyte gene expression in differentiation and syndromes of obesity/diabetes. J. Biol. Chem. 268: 6823–6826.

    PubMed  CAS  Google Scholar 

  • Tahara K, Aso H, Yamasaki T, Rose MT, Takasuga A, Sugimoto Y, Yamaguchi T, Tahara K, and Takano S (2004) Cloning and expression of type XII collagen isoforms during bovine adipogenesis. Differentiation. 72: 113–122.

    Article  PubMed  CAS  Google Scholar 

  • Van Barneveld RJ (2003) Modern pork production — Balancing efficient growth and feed conversion with product quality requirements and consumer demands. Asia Pac. J. Clin. Nutr. 12Suppl: S31.

    PubMed  Google Scholar 

  • Van Roermund CW, Hettema EH, Kal AJ, Van den Berg M, Tabak HF, and Wanders RJ (1998) Peroxisomal betaoxidation of polyunsaturated fatty acids in Saccharomyces cerevisiae: isocitrate dehydrogenase provides NADPH for reduction of double bonds at even positions. EMBO J. 17: 677–687.

    Article  PubMed  Google Scholar 

  • Wang Y and Watford M (2007) Glutamine, insulin and glucocorticoids regulate glutamine synthetase expression in C2C12 myotubes, Hep G2 hepatoma cells and 3T3 L1 adipocytes. Biochim. Biophys. Acta. 1770: 594–600.

    Article  PubMed  CAS  Google Scholar 

  • Wernersson R, Schieru M, Jørgensen F, Gorodkin J, Panitz F, Staerfeldt H, Christensen O, Mailund T, Hornshøj H, Klein A, et al. (2005) Pigs in sequence space: a 0.66X coverage pig genome survey based on shotgun sequencing. BMC Genomics. 10: 70.

    Article  Google Scholar 

  • Yan J, Burman A, Nichols C, Alila L, Showe LC, Showe MK, Boyer BB, Barnes BM, and Marr TG (2006) Detection of differential gene expression in brown adipose tissue of hibernating arctic ground squirrels with mouse microarrays. Physiol. Genomics. 25: 346–353.

    Article  PubMed  CAS  Google Scholar 

  • Yang DS, Smith JD, Zhou Z, Gandy SE, and Martins RN (1997) Characterization of the binding of amyloid-beta peptide to cell culture-derived native apolipoprotein E2, E3, and E4 isoforms and to isoforms from human plasma. J. Neurochem. 68: 721–725.

    Article  PubMed  CAS  Google Scholar 

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Authors and Affiliations

  1. Department of Biotechnology, Yeungnam University, Gyeongsan, 712-749, Korea

    Prati Bajracharya, Eun Jeong Jang & Inho Choi

  2. School of Biotechnology, Yeungnam University, Gyeongsan, 712-749, Korea

    Dong-Mok Lee & Eun Ju Lee

  3. Research Institute of GnC Bio, Daejeon, 305-150, Korea

    Sung Hwa Chae

  4. Bovine Resource Center, Korea Research Institute of Bioscience and Biotechnology, Daejeon, 305-806, Korea

    Hong-Seog Park

  5. Department of Parasitology, College of Medicine and UHRC, Inje University, Busan, 621-749, Korea

    Se Won Kang & Yong Seok Lee

  6. Department of Biochemistry and Molecular Biology, Eulji Medical College, Daejeon, 301-832, Korea

    Ki Ho Lee

  7. Department of Agricultural Sciences, Korea National Open University, Seoul, 110-791, Korea

    Jongsoo Chang

  8. Department of Animal Science, Chungbuk National University, Cheongju, 361-763, Korea

    Chung Soo Chung

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  1. Dong-Mok Lee
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  12. Inho Choi
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Correspondence to Inho Choi.

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Lee, DM., Bajracharya, P., Jang, E.J. et al. Gene expression profiles analyzed by DNA sequencing of cDNA clones constructed from porcine preadipocytes and adipocytes. Genes Genom 34, 125–131 (2012). https://doi.org/10.1007/s13258-011-0075-4

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  • DOI: https://doi.org/10.1007/s13258-011-0075-4

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