Journal of Comparative Physiology B

, Volume 180, Issue 5, pp 685–694 | Cite as

Gene expression survey of mitochondrial uncoupling proteins (UCP1/UCP3) in gilthead sea bream (Sparus aurata L.)

  • Azucena Bermejo-Nogales
  • Josep Alvar Calduch-Giner
  • Jaume Pérez-SánchezEmail author
Original Paper


The aim of this work is to underline the biological significance of mitochondrial uncoupling proteins (UCPs) in ectothermic fish using the gilthead sea bream (Sparus aurata L.) as an experimental model. A contig of 1,990 bp in length was recognized as a UCP1 ortholog after initial searches in the gilthead sea bream AQUAFIRST database ( Additional searches were performed in skeletal muscle by RT-PCR, and the amplified PCR product was recognized as UCP3 after sequence completion by 5′- and 3′RACE. UCP1 expression was mostly detected in liver, whereas UCP3 transcripts were only found in skeletal and cardiac muscle fibres (white skeletal muscle > red skeletal muscle > heart). Specific gene regulation of UCP1 (liver) and UCP3 (white skeletal muscle) was addressed in physiological models of age, seasonal growth and energy-metabolic unbalances. Both the increase in energy demand (stress confinement) and the reduction in energy supply during adaptive cold response in winter down-regulated UCP1 expression. Conversely, transcript levels of UCP3 were higher with age, seasonal fattening and dietary deficiencies in essential fatty acids leading to the increase in fatty acid flux towards the muscle. This close association between UCP1 and UCP3 with the oxidative and metabolic tissue status is perhaps directly related to the ancestral protein UCP function, and allows the use of UCPs as lipotoxicity markers in ectothermic fish.


Age Season Fatty acids Stress Lipotoxicity 



The authors are grateful to E. Rial and anonymous reviewers for helpful suggestions and revision of the manuscript and M.A. González for excellent technical assistance in PCR assays. This work was partially funded by the European Union (FOOD-CT-2006-16249: Sustainable Aquafeeds to Maximise the Health Benefits of Farmed Fish for Consumers, AQUAMAX) and Spanish (Ingenio-2010 Programme; Improvement of Aquaculture Production by the use of biotechnological tools, AQUAGENOMICS) projects. AB-N was recipient of a PhD fellowship from CSIC-BANCAJA.


  1. Abe T, Mujahid A, Sato K, Akiba Y, Toyomizu M (2006) Possible role of avian uncoupling protein in down-regulating mitochondrial superoxide production in skeletal muscle of fasted chickens. FEBS Lett 580:4815–4822CrossRefPubMedGoogle Scholar
  2. Benedito-Palos L, Saera-Vila A, Calduch-Giner JA, Kaushik S, Pérez-Sánchez J (2007) Combined replacement of fish meal and oil in practical diets for fast growing juveniles of gilthead sea bream (Sparus aurata L.): networking of systemic and local components of GH/IGF axis. Aquaculture 267:199–212CrossRefGoogle Scholar
  3. Benedito-Palos L, Navarro JC, Sitjà-Bobadilla A, Bell JG, Kaushik S, Pérez-Sánchez J (2008) High levels of vegetable oils in plant protein-rich diets fed to gilthead sea bream (Sparus aurata L.): growth performance, muscle fatty acid profiles and histological alterations of target tissues. Br J Nutr 100:992–1003CrossRefPubMedGoogle Scholar
  4. Bermejo-Nogales A, Benedito-Palos L, Saera-Vila A, Calduch-Giner JA, Sitjà-Bobadilla A, Pérez-Sánchez J (2007) Confinement exposure induces glucose regulated protein 75 (GRP75/mortalin/mtHsp70/PBP74/HSPA9B) in the hepatic tissue of gilthead sea bream (Sparus aurata L.). Comp Biochem Physiol 149B:428–438Google Scholar
  5. Björnsson BT, Johansson V, Benedet S, Einarsdottir IE, Hildahl J, Agustsson T, Jönsson E (2002) Growth hormone endocrinology of salmonids: regulatory mechanisms and mode of action. Fish Physiol Biochem 27:227–242CrossRefGoogle Scholar
  6. Boss O, Samec S, Paoloni-Giacobino A, Rossier C, Dulloo A, Seydoux J, Muzzin P, Giacobino JP (1997) Uncoupling protein-3: a new member of the mitochondrial carrier family with tissue-specific expression. FEBS Lett 408:39–42CrossRefPubMedGoogle Scholar
  7. Calduch-Giner JA, Mingarro M, Vega-Rubín de Celis S, Boujard D, Pérez-Sánchez J (2003) Molecular cloning and characterization of gilthead sea bream, (Sparus aurata) growth hormone receptor (GHR). Assessment of alternative splicing. Comp Biochem Physiol 136B:1–13Google Scholar
  8. Choi CS, Fillmore JJ, Kim JK, Liu ZX, Kim S, Collier EF, Kulkarni A, Distefano A, Hwang YJ, Kahn M, Chen Y, Yu CL, Moore IK, Reznick RM, Higashimori T, Shulman GI (2007) Overexpression of uncoupling protein 3 in skeletal muscle protects against fat-induced insulin resistance. J Clin Invest 117:1995–2003CrossRefPubMedGoogle Scholar
  9. Clapham JC, Arch JRS, Chapman H, Haynes A, Lister C, Moore GBT, Piercy V, Carter SA, Lehner I, Smith SA, Beeley LJ, Godden RJ, Herrity N, Skehel M, Changani KK, Hockings PD, Reid DG, Squires SM, Hatcher J, Trail B, Latcham J, Rastan S, Harper AJ, Cadenas S, Buckingham JA, Brand MD, Abuin A (2000) Mice overexpressing human uncoupling protein-3 in skeletal muscle are hyperphagic and lean. Nature 406:415–418CrossRefPubMedGoogle Scholar
  10. Emre Y, Hurtaud C, Ricquier D, Bouillaud F, Hughes J, Criscuolo F (2007) Avian UCP: the killjoy in the evolution of the mitochondrial uncoupling proteins. J Mol Evol 65:392–402CrossRefPubMedGoogle Scholar
  11. Evans ZP, Ellett JD, Schmidt MG, Schnellmann RG, Chavin KD (2008) Mitochondrial uncoupling protein-2 mediates steatotic liver injury following ischemia/reperfusion. J Biol Chem 283:8573–8579CrossRefPubMedGoogle Scholar
  12. Gates AC, Bernal-Mizrachi C, Chinault SL, Feng C, Schneider JG, Coleman T, Malone JP, Townsend RR, Chakravarthy MV, Semenkovich CF (2007) Respiratory uncoupling in skeletal muscle delays death and diminishes age-related disease. Cell Metab 6:497–505CrossRefPubMedGoogle Scholar
  13. Gonzalez-Barroso MD, Ricquier D, Cassard-Doulcier AM (2000) The human uncoupling protein-1 gene (UCP1): present status and perspectives in obesity research. Obes Rev 1:61–72CrossRefGoogle Scholar
  14. Harper JA, Stuart JA, Jekabsons MB, Roussel D, Brindle KM, Dickinson K, Jones RB, Brand MD (2002) Artifactual uncoupling by uncoupling protein 3 in yeast mitochondria at the concentrations found in mouse and rat skeletal-muscle mitochondria. Biochem J 361:49–56CrossRefPubMedGoogle Scholar
  15. Heaton GM, Wagenvoord RJ, Kemp A, Nicholls DG (1978) Brown adipose tissue mitochondria: photoaffinity labelling of the regulatory site of energy dissipation. Eur J Biochem 82:515–521CrossRefPubMedGoogle Scholar
  16. Hesselink MKC, Keizer HA, Borghouts LB, Schaart G, Kornips CFP, Slieker LJ, Sloop KW, Saris WHM, Schrauwen P (2001) Protein expression of UCP3 differs between human type 1, type 2a, and type 2b fibers. FASEB J 15:1071–1073PubMedGoogle Scholar
  17. Hoeks J, Hesselink MKC, van Bilsen M, Schaart G, van der Vusse GJ, Saris WHM, Schrauwen P (2003) Differential response of UCP3 to medium versus long chain triacylglycerols; manifestation of a functional adaptation. FEBS Lett 555:631–637CrossRefPubMedGoogle Scholar
  18. Horvath T, Diano S, Miyamoto S, Barry S, Gatti S, Alberati D, Livak F, Lombardi A, Moreno M, Goglia F, Mor G, Hamilton J, Kachinskas D, Horwitz B, Warden CH (2003) Uncoupling proteins-2 and 3 influence obesity and inflammation in transgenic mice. Int J Obes 27:433–442CrossRefGoogle Scholar
  19. Hughes J, Criscuolo F (2008) Evolutionary history of the UCP gene family: gene duplication and selection. BMC Evol Biol 8:306CrossRefPubMedGoogle Scholar
  20. Hughes DA, Jastroch M, Stoneking M, Klingenspor M (2009) Molecular evolution of UCP1 and the evolution history of mammalian non-shivering thermogenesis. BMC Evol Biol 9:4CrossRefPubMedGoogle Scholar
  21. Jastroch M, Wuertz S, Kloas W, Klingenspor M (2005) Uncoupling protein 1 in fish uncovers an ancient evolutionary history of mammalian nonshivering thermogenesis. Physiol Genomics 22:150–156CrossRefPubMedGoogle Scholar
  22. Jastroch M, Buckingham JA, Helwig M, Klingenspor M, Brand MD (2007) Functional characterisation of UCP1 in the common carp: uncoupling activity in liver mitochondria and cold-induced expression in the brain. J Comp Physiol 177B:743–752Google Scholar
  23. Jastroch M, Withers KW, Taudien S, Frappell PB, Helwig M, Fromme T, Hirschberg V, Heldmaier G, McAllan BM, Firth BT, Burmester T, Platzer M, Klingenspor M (2008) Marsupial uncoupling protein 1 sheds light on the evolution of mammalian nonshivering thermogenesis. Physiol Genomics 32:161–169PubMedGoogle Scholar
  24. Jiang N, Zhang G, Bo H, Qu J, Ma G, Cao D, Wen L, Liu S, Ji LL, Zhang Y (2009) Upregulation of uncoupling protein-3 in skeletal muscle during exercise: a potential antioxidant function. Free Radic Biol Med 46:138–145CrossRefPubMedGoogle Scholar
  25. Kontani Y, Wang Y, Kimura K, Inokuma KI, Saito M, Suzuki-Miura T, Wang Z, Sato Y, Mori N, Yamashita H (2005) UCP1 deficiency increases susceptibility to diet-induced obesity with age. Aging Cell 4:147–155CrossRefPubMedGoogle Scholar
  26. Kozak LP, Koza RA (1999) Mitochondrial uncoupling proteins and obesity: molecular and genetic aspects of UCP1. Int J Obes 23:S33–S37CrossRefGoogle Scholar
  27. Kratky D, Strauss JG, Zechner R (2001) Tissue-specific activity of lipoprotein lipase in skeletal muscle regulates the expression of uncoupling protein 3 in transgenic mouse models. Biochem J 355:647–652PubMedGoogle Scholar
  28. Krauss S, Zhang CY, Lowell BB (2005) The mitochondrial uncoupling-protein homologues. Nat Rev Mol Cell Biol 6:248–261CrossRefPubMedGoogle Scholar
  29. Liang XF, Ogata HY, Oku H, Chen J, Hwang F (2003) Abundant and constant expression of uncoupling protein 2 in the liver of red sea bream Pagrus major. Comp Biochem Physiol 136A:655–661Google Scholar
  30. Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2−ΔΔCT method. Methods 25:402–408CrossRefPubMedGoogle Scholar
  31. Locke RM, Rial E, Nicholls DG (1982) The acute regulation of mitochondrial proton conductance in cells and mitochondria from the brown fat of cold-adapted and warm-adapted guinea pigs. Eur J Biochem 129:381–387CrossRefPubMedGoogle Scholar
  32. Lynn SG, Powell KA, Westneats DF, Shepherd BS (2009) Seasonal and sex-specific mRNA levels of key endocrine genes in adult yellow perch (Perca flavescens) from Lake Erie. Mar Biotechnol 11:210–222CrossRefPubMedGoogle Scholar
  33. Mark FC, Lucassen M, Pörtner HO (2006) Thermal sensitivity of uncoupling protein expression in polar and temperate fish. Comp Biochem Physiol 1D:365–374Google Scholar
  34. Mingarro M, Vega-Rubín de Celis R, Astola A, Pendón C, Valdivia MM, Pérez-Sánchez J (2002) Endocrine mediators of seasonal growth in gilthead sea bream (Sparus aurata): the growth hormone and somatolactin paradigm. Gen Comp Endocrinol 128:102–111CrossRefPubMedGoogle Scholar
  35. Modrianský M, Murdza-Inglis DL, Patel HV, Freeman KB, Garlid KD (1997) Identification by site-directed mutagenesis of three arginines in uncoupling protein that are essential for nucleotide binding and inhibition. J Biol Chem 272:24759–24762CrossRefPubMedGoogle Scholar
  36. Nabben M, Hoeks J (2008) Mitochondrial uncoupling protein 3 and its role in cardiac- and skeletal muscle metabolism. Physiol Behav 94:259–269CrossRefPubMedGoogle Scholar
  37. Nabben M, Hoeks J, Briedé JJ, Glatz JFC, Moonen-Kornips E, Hesselink MKC, Schrauwen P (2008) The effect of UCP3 overexpression on mitochondrial ROS production in skeletal muscle of young versus aged mice. FEBS Lett 582:4147–4152CrossRefPubMedGoogle Scholar
  38. Nicholls DG, Locke RM (1984) Thermogenic mechanisms in brown fat. Physiol Rev 64:1–64PubMedGoogle Scholar
  39. Parker N, Crichton PG, Vidal-Puig AJ, Brand MD (2009) Uncoupling protein-1 (UCP1) contributes to the basal proton conductance of brown adipose tissue mitochondria. J Bioenerg Biomembr 41:335–342CrossRefPubMedGoogle Scholar
  40. Pecqueur C, Cassard-Doulcier AM, Raimbault S, Miroux B, Fleury C, Gelly C, Bouillaud F, Ricquier D (1999) Functional organization of the human uncoupling protein-2 gene, and juxtaposition to the uncoupling protein-3 gene. Biochem Biophys Res Commun 255:40–46CrossRefPubMedGoogle Scholar
  41. Pecqueur C, Alves-Guerra MC, Gelly C, Lévi-Meyrueis C, Couplan E, Collins S, Ricquier D, Bouillaud F, Miroux B (2001) Uncoupling protein 2, in vivo distribution, induction upon oxidative stress, and evidence for translational regulation. J Biol Chem 276:8705–8712CrossRefPubMedGoogle Scholar
  42. Pérez-Sánchez J, Calduch-Giner JA, Mingarro M, Vega-Rubín de Celis S, Gómez-Requeni P, Saera-Vila A, Astola A, Valdivia MM (2002) Overview of fish growth hormone family. New insights in genomic organization and heterogeneity of growth hormone receptors. Fish Physiol Biochem 27:243–258CrossRefGoogle Scholar
  43. Peterson JM, Bryner RW, Frisbee JC, Alway SE (2008) Effects of exercise and obesity on UCP3 content in rat hindlimb muscles. Med Sci Sports Exerc 40:1616–1622CrossRefPubMedGoogle Scholar
  44. Raimbault S, Dridi S, Denjean F, Lachuer J, Couplan E, Bouillaud F, Bordas A, Duchamp C, Taouis M, Ricquier D (2001) An uncoupling protein homologue putatively involved in facultative muscle thermogenesis in birds. Biochem J 353:441–444CrossRefPubMedGoogle Scholar
  45. Rial E, Poustie A, Nicholls DG (1983) Brown adipose tissue mitochondria: regulation of the 32000-Mr uncoupling protein by fatty acids and purine nucleotides. Eur J Biochem 137:197–203CrossRefPubMedGoogle Scholar
  46. Saera-Vila A, Calduch-Giner JA, Navarro I, Pérez-Sánchez J (2007) Tumour necrosis factor (TNF)α as a regulator of fat tissue mass in the Mediterranean gilthead sea bream (Sparus aurata L.). Comp Biochem Physiol 146B:338–345Google Scholar
  47. Samec S, Seydoux J, Russell AP, Montani JP, Dulloo AG (2002) Skeletal muscle heterogeneity in fasting-induced upregulation of genes encoding UCP2, UCP3, PPARγ and key enzymes of lipid oxidation. Pflugers Arch 445:80–86CrossRefPubMedGoogle Scholar
  48. Solanes G, Vidal-Puig A, Grujic D, Flier JS, Lowell BB (1997) The human uncoupling protein-3 gene. Genomic structure, chromosomal localization, and genetic basis for short and long form transcripts. J Biol Chem 272:25433–25436CrossRefPubMedGoogle Scholar
  49. Stuart JA, Harper KM, Brindle KM, Brand MD (1999) Uncoupling protein 2 from carp and zebrafish, ectothermic vertebrates. Biochim Biophys Acta 1413:50–54CrossRefPubMedGoogle Scholar
  50. Vidal-Puig A, Solanes G, Grujic D, Flier JS, Lowell BB (1997) UCP3: an uncoupling protein homologue expressed preferentially and abundantly in skeletal muscle and brown adipose tissue. Biochem Biophys Res Commun 235:79–82CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag 2010

Authors and Affiliations

  • Azucena Bermejo-Nogales
    • 1
  • Josep Alvar Calduch-Giner
    • 1
  • Jaume Pérez-Sánchez
    • 1
    Email author
  1. 1.Fish Nutrition and Growth Endocrinology Group, Department of Biology, Culture and Pathology of Marine Fish SpeciesInstitute of Aquaculture Torre de la Sal (IATS), CSICRibera de CabanesSpain

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