Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Paper
  • Published:

Uncoupling proteins-2 and 3 influence obesity and inflammation in transgenic mice

Abstract

OBJECTIVE: To test the hypothesis that either uncoupling protein-2 UCP2 or UCP3 or both together influence obesity and inflammation in transgenic mice.

DESIGN: We generated 12 lines of transgenic mice for both human UCP2 and 3 using native promoters from a human bacterial artificial chromosome (BAC) clone. The BAC expresses no genes other than UCP2 and 3. Mice used for experiments are N4 or higher of backcross to C57BL/6J (B6). Each experiment used transgenic mice and their nontransgenic littermates.

RESULTS: Northern blots confirmed expression on human UCP2 in adipose and spleen, while human UCP3 expression was detectable in gastrocnemius muscle. Western blots demonstrated a four-fold increase of UCP2 protein in spleens of Line 32 transgenic animals. Heterozygous mice of four lines showing expression of human UCP2 in spleen were examined for obesity phenotypes. There were no significant differences between Lines 1 and 32, but female transgenics of both lines had significantly smaller femoral fat depots than the control (littermate) mice (P=0.015 and 0.005, respectively). In addition, total fat of transgenic females was significantly less in Line 1 (P=0.05) and almost significantly different in Line 32 (P=0.06). Male Line 1 mice were leaner (P=0.04) while male Line 32 mice were almost significantly leaner (P=0.06). Heterozygous mice of Lines 35 and 44 showed no significant differences from the nontransgenic littermate controls. Effects of the UCP2/UCP3 transgene on obesity in Line 32 mice were confirmed by crossing transgenic mice with the B6.Cg-Ay agouti obese mice. B6.Cg-Ay carrying the UCP2/UCP3 transgene from Line 32 were significantly leaner than nontransgenic B6.Cg-Ay mice.

Line 32 UCP2/UCP3 transgenics showed increased hypothalamic Neuropeptide (NPY) levels and food intake, with reduced spontaneous physical activity. Transgenic baseline interleukin4 (IL-4) and interleukin6 (IL-6) levels were low with lower or later increases after endotoxin injection compared to wild-type littermates. Endotoxin-induced fever was also diminished in transgenic male animals. Low-density lipoprotein (LDL) cholesterol levels were significantly higher in both Line 1 and 32 transgenics (P=0.05 and 0.001, respectively) after they had been placed on a moderate fat-defined diet containing 32% of calories from fat for 5 weeks.

CONCLUSION: Moderate overexpression of UCP2 and 3 reduced fat mass and increased LDL cholesterol in two independent lines of transgenic mice. Thus, the reduced fat mass cannot be due to insertional mutagenesis since virtually identical fat pad weights and masses were observed with the two independent lines. Line 32 mice also have altered inflammation and mitochondrial function. We conclude that UCP2 and/or 3 have small but significant effects on obesity in mice, and that their mechanism of action may include alterations of metabolic rate.

This is a preview of subscription content, access via your institution

Access options

Rent or buy this article

Prices vary by article type

from$1.95

to$39.95

Prices may be subject to local taxes which are calculated during checkout

Figure 1
Figure 2
Figure 3
Figure 4

Similar content being viewed by others

References

  1. Nicholls DG, Locke RM . Thermogenic mechanisms in brown fat. Physiol Rev 1984; 64: 1–64.

    Article  CAS  PubMed  Google Scholar 

  2. Bouillaud F, Ricquier D, Thibault J, Weissenbach J . Molecular approach to thermogenesis in brown adipose tissue: cDNA cloning of the mitochondrial uncoupling protein. Proc Nat Acad Sci USA 1985; 82: 445–448.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  3. Fleury C, Neverova M, Collins S, Raimbault S, Champigny O, Levi-Meyrueis C, Bouillaud F, Seldin MF, Surwit RS, Ricquier D, Warden CH . Uncoupling protein-2: a novel gene linked to obesity and hyperinsulinemia [see comments]. Nat Genet 1997; 15: 269–272.

    Article  CAS  PubMed  Google Scholar 

  4. Boss O, Samec S, Paoloni-Giacobino A, Rossier C, Dulloo A, Seydoux J, Muzzin P, Giacobino JP . Uncoupling protein-3: a new member of the mitochondrial carrier family with tissue-specific expression. FEBS Lett 1997; 408: 39–42.

    Article  CAS  PubMed  Google Scholar 

  5. Vidal-Puig A, Solanes G, Grujic D, Flier JS, Lowell BB . UCP3: an uncoupling protein homologue expressed preferentially and abundantly in skeletal muscle and brown adipose tissue. Biochem Biophys Res Commun 1997; 235: 79–82.

    Article  CAS  PubMed  Google Scholar 

  6. Mao W, Yu XX, Zhong A, Li W, Brush J, Sherwood SW, Adams SH, Pan G . UCP4, a novel brain-specific mitochondrial protein that reduces membrane potential in mammalian cells. FEBS Lett 1999; 443: 326–330.

    Article  CAS  PubMed  Google Scholar 

  7. Sanchis D, Fleury C, Chomiki N, Goubern M, Huang Q, Neverova M, Gregoire F, Easlick J, Raimbault S, Levi-Meyrueis C, Miroux B, Collins S, Seldin M, Richard D, Warden C, Bouillaud F, Ricquier D . BMCP1, a novel mitochondrial carrier with high expression in the central nervous system of humans and rodents, and respiration uncoupling activity in recombinant yeast. J Biol Chem 1998; 273: 34611–34615.

    Article  CAS  PubMed  Google Scholar 

  8. Lowell BB, Spiegelman BM . Towards a molecular understanding of adaptive thermogenesis. Nature 2000; 404: 652–660.

    Article  CAS  PubMed  Google Scholar 

  9. Richard D, Rivest R, Huang Q, Bouillaud F, Sanchis D, Champigny O, Ricquier D . Distribution of the uncoupling protein 2 mRNA in the mouse brain. J Comp Neurol 1998; 397: 549–560.

    Article  CAS  PubMed  Google Scholar 

  10. Horvath TL, Warden CH, Hajos M, Lombardi A, Goglia F, Diano S . Brain uncoupling protein 2: uncoupled neuronal mitochondria predict thermal synapses in homeostatic centers. J Neurosci 1999; 19: 10417–10427.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  11. Diano S, Urbanski HF, Horvath B, Bechmann I, Kagiya A, Nemeth G, Naftolin F, Warden CH, Horvath TL . Mitochondrial uncoupling protein 2 (UCP2) in the nonhuman primate brain and pituitary. Endocrinology 2000; 141: 4226–4238.

    Article  CAS  PubMed  Google Scholar 

  12. Esterbauer H, Schneitler C, Oberkofler H, Ebenbichler C, Paulweber B, Sandhofer F, Ladurner G, Hell E, Strosberg AD, Patsch JR, Krempler F, Patsch W . A common polymorphism in the promoter of UCP2 is associated with decreased risk of obesity in middle-aged humans. Nat Genet 2001; 28: 178–183.

    Article  CAS  PubMed  Google Scholar 

  13. Schonfeld-Warden NA, Warden CH . Physiological effects of variants in human uncoupling proteins: UCP2 influences body-mass index. Biochem Soc Trans 2001; 29: 777–784.

    Article  CAS  PubMed  Google Scholar 

  14. Arsenijevic D, Onuma H, Pecqueur C, Raimbault S, Manning BS, Miroux B, Couplan E, Alves-Guerra MC, Goubern M, Surwit R, Bouillaud F, Richard D, Collins S, Ricquier D . Disruption of the uncoupling protein-2 gene in mice reveals a role in immunity and reactive oxygen species production. Nat Genet 2000; 26: 435–439.

    Article  CAS  PubMed  Google Scholar 

  15. Zhang CY, Baffy G, Perret P, Krauss S, Peroni O, Grujic D, Hagen T, Vidal-Puig AJ, Boss O, Kim YB, Zheng XX, Wheeler MB, Shulman GI, Chan CB, Lowell BB . Uncoupling protein-2 negatively regulates insulin secretion and is a major link between obesity, beta cell dysfunction, and type 2 diabetes. Cell 2001; 105: 745–755.

    Article  CAS  PubMed  Google Scholar 

  16. Vidal-Puig AJ, Grujic D, Zhang CY, Hagen T, Boss O, Ido Y, Szczepanik A, Wade J, Mootha V, Cortright R, Muoio DM, Lowell BB . Energy metabolism in uncoupling protein 3 gene knockout mice. J Biol Chem 2000; 275: 16258–16266.

    Article  CAS  PubMed  Google Scholar 

  17. Gong DW, Monemdjou S, Gavrilova O, Leon LR, Marcus-Samuels B, Chou CJ, Everett C, Kozak LP, Li C, Deng C, Harper ME, Reitman ML . Lack of obesity and normal response to fasting and thyroid hormone in mice lacking uncoupling protein-3. J Biol Chem 2000; 275: 16251–16257.

    Article  CAS  PubMed  Google Scholar 

  18. Clapham JC, Arch JR, Chapman H, Haynes A, Lister C, Moore GB, 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 . Mice overexpressing human uncoupling protein-3 in skeletal muscle are hyperphagic and lean. Nature 2000; 406: 415–418.

    Article  CAS  PubMed  Google Scholar 

  19. Monemdjou S, Hofmann WE, Kozak LP, Harper ME . Increased mitochondrial proton leak in skeletal muscle mitochondria of UCP1-deficient mice. Am J Physiol Endocrinol Metab 2000; 279: E941–E946.

    Article  CAS  PubMed  Google Scholar 

  20. Kozak LP, Koza RA . Mitochondria uncoupling proteins and obesity: molecular and genetic aspects of UCP1. Int J Obes Relat Metab Disord 1999; 23(Suppl 6): S33–S37.

    Article  CAS  PubMed  Google Scholar 

  21. Horwitz BA, Hamilton JS, Routh VH, Green K, Havel P, Chan A . Adiposity and serum leptin increase in fatty (fa/fa) BNZ neonates without decreased VMH serotonergic activity. Am J Physiol 1998; 274: E1009–E1017.

    CAS  PubMed  Google Scholar 

  22. Chai Z, Gatti S, Toniatti C, Poli V, Bartfai T . Interleukin (IL)-6 gene expression in the central nervous system is necessary for fever response to lipopolysaccharide or IL-1 beta: a study on IL-6-deficient mice. J Exp Med 1996; 183: 311–316.

    Article  CAS  PubMed  Google Scholar 

  23. Lombardi A, Lanni A, Moreno M, Brand MD, Goglia F . Effect of 3,5-di-iodo-L-thyronine on the mitochondrial energy-transduction apparatus. Biochem J 1998; 330(Part 1): 521–526.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  24. Horvath TL, Kalra SP, Naftolin F, Leranth C . Morphological evidence for a galanin–opiate interaction in the rat mediobasal hypothalamus. J Neuroendocrinol 1995; 7: 579–588.

    Article  CAS  PubMed  Google Scholar 

  25. Negre-Salvayre A, Hirtz C, Carrera G, Cazenave R, Troly M, Salvayre R, Penicaud L, Casteilla L . A role for uncoupling protein-2 as a regulator of mitochondrial hydrogen peroxide generation. FASEB J 1997; 11: 809–815.

    Article  CAS  PubMed  Google Scholar 

  26. Gavrilova O, Leon LR, Marcus-Samuels B, Mason MM, Castle AL, Refetoff S, Vinson C, Reitman ML . Torpor in mice is induced by both leptin-dependent and -independent mechanisms. Proc Nat Acad Sci USA 1999; 96: 14623–14628.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  27. Kalra SP, Dube MG, Pu S, Xu B, Horvath TL, Kalra PS . Interacting appetite-regulating pathways in the hypothalamic regulation of body weight. Endocr Rev 1999; 20: 68–100.

    CAS  PubMed  Google Scholar 

  28. Warden CH, Fisler JS, Pace MJ, Svenson KL, Lusis AJ . Coincidence of genetic loci for plasma cholesterol levels and obesity in a multifactorial mouse model. J Clin Invest 1993; 92: 773–779.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  29. Cadenas S, Echtay KS, Harper JA, Jekabsons MB, Buckingham JA, Chapman H, Clapham JC, Brand MD . The basal proton conductance of skeletal muscle mitochondria from transgenic mice overexpressing or lacking uncoupling protein-3. J Biol Chem 2002; 277: 2773–2778.

    Article  CAS  PubMed  Google Scholar 

  30. Garcia-Martinez C, Sibille B, Solanes G, Darimont C, Mace K, Villarroya F, Gomez-Foix AM . Overexpression of UCP3 in cultured human muscle lowers mitochondrial membrane potential, raises ATP/ADP ratio, and favors fatty acid vs glucose oxidation. FASEB J 2001; 15: 2033–2035.

    Article  CAS  PubMed  Google Scholar 

  31. Short KR, Nygren J, Barazzoni R, Levine J, Nair KS . T(3) increases mitochondrial ATP production in oxidative muscle despite increased expression of UCP2 and -3. Am J Physiol Endocrinol Metab 2001; 280: E761–E769.

    Article  CAS  PubMed  Google Scholar 

  32. Meirhaeghe A, Amouyel P, Helbecque N, Cottel D, Otabe S, Froguel P, Vasseur F . An uncoupling protein 3 gene polymorphism associated with a lower risk of developing type II diabetes and with atherogenic lipid profile in a French cohort. Diabetologia 2000; 43: 1424–1428.

    Article  CAS  PubMed  Google Scholar 

  33. Warden CH, Fisler JS, Shoemaker SM, Wen PZ, Svenson KL, Pace MJ, Lusis AJ . Identification of four chromosomal loci determining obesity in a multifactorial mouse model. J Clin Invest 1995; 95: 1545–1552.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  34. Li B, Nolte LA, Ju JS, Han DH, Coleman T, Holloszy JO, Semenkovich CF . Skeletal muscle respiratory uncoupling prevents diet-induced obesity and insulin resistance in mice. Nat Med 2000; 6: 1115–1120.

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

This work was supported by grants from the National Institute of Diabetes and Kidney Disease (DK53993), National Institute of Mental Health (MH-59847), National Institute of Research Resources (RR-14451), the National Institute for Neurological Disorders and Stroke (NS-41725) and the Wyeth-Ayerst Research Institute.

Author information

Authors and Affiliations

Authors

Corresponding authors

Correspondence to T L Horvath or C H Warden.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Horvath, T., Diano, S., Miyamoto, S. et al. Uncoupling proteins-2 and 3 influence obesity and inflammation in transgenic mice. Int J Obes 27, 433–442 (2003). https://doi.org/10.1038/sj.ijo.0802257

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/sj.ijo.0802257

Keywords

This article is cited by

Search

Quick links