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Development of a Porcine cDNA Microarray: Analysis of Clenbuterol Responding Genes in Pig (Sus scrofa) Internal Organs |
ZHANG Jin, GUO Wei, SHEN Liang-cai, LIU Qiu-yue, DENG Xue-mei, HU Xiao-xiang , LI Ning |
1.College of Life Science and Biotechnology, Hebei Normal University of Science & Technology, Qinhuangdao 066600, P.R.China
2.State Key Laboratory for Agrobiotechnology, China Agricultural University, Beijing 100193, P.R.China
3.College of Animal Science and Technology, China Agricultural University, Beijing 100193, P.R.China |
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摘要 Pig (Sus scrofa) fat accumulation can be reduced by feeding with high dosages of clenbuterol, but the molecular mechanism has not yet been explained. In our study, a porcine cDNA microarray representing 3 358 pig genes was successfully developed. This microarray is the first porcine DNA microarray in China and its false positive rate is 0.98%, which means the microarray platform is reliable. The microarray can be used to study gene expression profiles in multiple pig tissues because the present genes percentage of adipose, skeletal muscle, heart, liver, lung, kidney, and spleen were all more than 60%. This microarray was used to identify the genes responding to clenbuterol stimulation in pig internal organs, including heart, liver, lung, spleen, and kidney. Many genes were identified including enzymes involved in lipids metabolism (lipoprotein lipase up-regulated in liver, heart and lung, ATP-citrate lyase and carnitine palmitoyltransferase II precursor up-regulated in liver, succinyl-CoA up-regulated in lung, mitochondrial malate dehydrogenase down-regulated in spleen), and signaling pathway genes (cAMP-protein kinase A signaling pathway was found up-regulated in liver, heart, lung, and kidney as reported previously, while transforming growth factor was found down-regulated in heart and lung). However, no common gene responding to clenbuterol administration was found in all tissues. The expression levels of 14 genes were analyzed using real-time PCR with 82.1% of them induced to express similar magnitudes as in the microarray analyses. This work offers some understanding of how clenbuterol so effectively reduces pig adipose accumulation on the molecular level.
Abstract Pig (Sus scrofa) fat accumulation can be reduced by feeding with high dosages of clenbuterol, but the molecular mechanism has not yet been explained. In our study, a porcine cDNA microarray representing 3 358 pig genes was successfully developed. This microarray is the first porcine DNA microarray in China and its false positive rate is 0.98%, which means the microarray platform is reliable. The microarray can be used to study gene expression profiles in multiple pig tissues because the present genes percentage of adipose, skeletal muscle, heart, liver, lung, kidney, and spleen were all more than 60%. This microarray was used to identify the genes responding to clenbuterol stimulation in pig internal organs, including heart, liver, lung, spleen, and kidney. Many genes were identified including enzymes involved in lipids metabolism (lipoprotein lipase up-regulated in liver, heart and lung, ATP-citrate lyase and carnitine palmitoyltransferase II precursor up-regulated in liver, succinyl-CoA up-regulated in lung, mitochondrial malate dehydrogenase down-regulated in spleen), and signaling pathway genes (cAMP-protein kinase A signaling pathway was found up-regulated in liver, heart, lung, and kidney as reported previously, while transforming growth factor was found down-regulated in heart and lung). However, no common gene responding to clenbuterol administration was found in all tissues. The expression levels of 14 genes were analyzed using real-time PCR with 82.1% of them induced to express similar magnitudes as in the microarray analyses. This work offers some understanding of how clenbuterol so effectively reduces pig adipose accumulation on the molecular level.
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Received: 02 June 2011
Accepted:
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Fund: This work was supported by the National Natural Science Foundation of China (30800778 and 31072004), the Hebei Natural Science Foundation (C2009000871), the Hebei Educational Foundation, China (2009119), the Hebei Excellent Expert for Overseas Advanced Training Program (2009), and Scientific Research Innovation Team Funds of Hebei Normal University of Sci & Tech, China. |
Corresponding Authors:
Correspondence HU Xiao-xiang, E-mail: huxx@cau.edu.cn; LI Ning, Tel: +86-10-62733323, Fax: +86-10-62733904, E-mail: ninglbau@public3.bta.net.cn
E-mail: huxx@cau.edu.cn
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Cite this article:
ZHANG Jin, GUO Wei, SHEN Liang-cai, LIU Qiu-yue, DENG Xue-mei, HU Xiao-xiang , LI Ning.
2012.
Development of a Porcine cDNA Microarray: Analysis of Clenbuterol Responding Genes in Pig (Sus scrofa) Internal Organs. Journal of Integrative Agriculture, 12(11): 1877-1883.
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[1]Band M R, Larson J H, Rebiez M, Green C A, Heyen D W,Donovan J, Windish R, Steining C, Mahyuddin P,Womack J E, et al. 2000. An ordered comparative mapof the cattle and human genomes. Genome Research,10, 1359-1367[2]Brown P, Botstein D. 1999. Exploring the new world of thegenome with DNA microarrays. Nature Genetics, 21,33-37[3]Egan J J, Greenberg A S, Chang M K, Wek S A, Moos Jr M .2012, CAAS. All rights reserved. Published by Elsevier Ltd.C, Londos C. 1992. Mechanism of hormone-stimulatedlipolysis in adipocytes: translocation of hormonesensitivelipase to the lipid storage droplet.Proceedings of the National Academy of Sciences ofUnited States of America, 89, 8537-8541[4]Gesta S, Tseng Y H, Kahn C R. 2007. Developmental originof fat: tracking obesity to its source. Cell, 131, 242-256[5]Greenberg A S, Egan J J, Wek S A, Garty N B, Blanchette-Mackie E J, Londos C. 1991. Perilipin a major hormonallyregulated adipocyte-specif ic phosphoproteinassociated with the periphery of lipid storage droplets.Journal of Biological Chemistry, 266, 11341-11346[6]Hoheisel J. 2006. Microarray technology: beyond transcriptprofiling and genotype analysis. Nature ReviewGenetics, 7, 200-210[7]Jares P. 2006. DNA microarray applications in functionalgenomics. Ultrastructural Pathology, 30, 209-219[8]Kuiper H A, Noordam M Y, van Dooren-Flipsen M M H,Schilt R, Roos A H. 1998. Illegal use of beta-adrenergicagonists: European Community. Journal AnimalScience, 76, 195-207[9]Li S, Li Y, Du W, Zhang L, Yu S, Dai Y, Zhao C, Li N. 2005.Aberrant gene expression in organs of bovine clonesthat die within two days after birth. Biology ofReproduction, 72, 258-265[10]Londos C, Honnor R C, Dhillon G S. 1985. cAMPdependentprotein kinase and lipolysis in rat adipocytesIII Multiple modes of insulin regulation of lipolysis andregulation of insulin responses by adenylate cyclaseregulators. Journal of Biological Chemistry, 260,15139-15145[11]Martinez-Navarro J F. 1990. Food poisoning related toconsumption of illicit ?-agonist in liver. The Lancet,336, 1311.[12]Mazzanti G, Daniele C, Boatto G, Manca G, Brambilla G,Loizzo A. 2003. New ?-adrenergic agonists used illicitlyas growth promoters in animal breeding: chemical andpharmacodynamic studies. Toxicology, 187, 91-99[13]Mitchell G A, Dunnavan G. 1998. Illegal use of betaadrenergicagonists in the United States. JournalAnimal Science, 76, 208-211[14]Plant D R, Kearns C F, McKeever K H, Lynch G S. 2003.Therapeutic clenbuterol treatment does not alter Ca2+sensitivity of permeabilized fast muscle fibres fromexercise trained or untrained horses. Journal of MuscleResearch and Cell Motility, 24, 471-476[15]Pospisilik J A, Schramek D, Schnidar H, Cronin S J, NehmeN T, Zhang X, Knauf C, Cani P D, Aumayr K, Todoric J,et al. 2010. Drosophila genome-wide obesity screenreveals hedgehog as a determinant of brown versuswhite adipose cell fate. Cell, 140, 148-160[16]Pulce C, Lamaison D, Keck G, Bostvironnois C, Nicolas J,Descotes J. 1991. Collective human food poisoning byclenbuterol residues in veal liver. Veterinary andHuman Toxicology, 33, 480-481[17]Sawada T, Miyoshi H, Shimada K, Suzuki A, Okamatsu-Ogura Y, Perfield J W, Kondo T, Nagai S, Shimizu C,Yoshioka N, et al. 2010. Perilipin overexpression in whiteadipose tissue induces a brown fat-like phenotype. PLoSOne, 5, e14006.Schena M. 2003. Microarray Analysis. Rochester, JohnWiley and Sons, NY.[18]Schena M, Shalon D, Davis R, Brown P. 1995. Quantitativemonitoring of gene expression patterns with acomplementary DNA microarray. Science, 270, 467-470[19]Shiu T C, Chong W H. 2001. A cluster of clenbuterolpoisoning associated with pork and pig offal in HongKong. Public Health and Epidemiology Bulletin, 10,14-17[20]Spurlock DM,McDonald T G,McIntyre LM. 2006. Changesin skeletal muscle gene expression following clenbuteroladministration. BMC Genomics, 7, 320.[21]Tendi E A, Cunsolo R, Bellia D, Messina R L, Paratore S,Calissano P, Cavallaro S. 2010. Drug target identificationfor neuronal apoptosis through a genome scalescreening. Current Medicinal Chemistry, 17, 2906-2920[22]Wang C S, McConathyW J, Kloer H U, Alaupovic P. 1985.Modulat ion of lipoprotein lipase activity byapolipoproteins. Effect of apolipoprotein C-III. Journalof Clinical Investigation, 75, 384-390[23]Wang H, Eckel R H. 2009. Lipoprotein lipase: from gene toobes i ty. Amer ican Journal of Phys iology -Endocrinology and Metabolism, 297, E271-E288.[24]Yang Y, Dudoit S, Luu P, Lin D, Peng V, Ngai J, Speed T.2002. Normalization for cDNA microarray data: a robustcomposite method addressing single and multiple slidesystematic variation. Nucleic Acids Research, 30, e15.[25]Zhang J, Hupfeld C J, Taylor S S, Olefsky J M, Tsien R Y.2005. Insulin disrupts beta-adrenergic signaling toprotein kinase A in adipocytes. Nature, 473, 569-573[26]Zhang J, He Q, Liu Q Y, Guo W, Deng X M, Zhang W W,Hu X X, Li N. 2007. Differential gene expression profilein pig adipose tissue treated with/without clenbuterol.BMC Genomics, 8, 433.[27]Zhou L, Li Y, Nie T, Feng S, Yuan J, Chen H, Yang Z. 2007.Clenbuterol inhibits SREBP-1c expression by activatingCREB1 Journal of Biochemistry and MolecularBiology, 40, 525-531. |
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