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.

  • Protocol
  • Published:

Cold-induced activation of brown adipose tissue and adipose angiogenesis in mice

Nature Protocols volume 7, pages 606–615 (2012)Cite this article

Subjects

Abstract

Exposure of humans and rodents to cold activates thermogenic activity in brown adipose tissue (BAT). This protocol describes a mouse model to study the activation of BAT and angiogenesis in adipose tissues by cold acclimation. After a 1-week exposure to 4 °C, adult C57BL/6 mice show an obvious transition from subcutaneous white adipose tissue (WAT) into brown-like adipose tissue (BRITE). The BRITE phenotype persists after continuous cold exposure, and by the end of week 5 BRITE contains a high number of uncoupling protein-1–positive mitochondria, a characteristic feature of BAT. During the transition from WAT into BRITE, the vascular density is markedly increased owing to the activation of angiogenesis. In BAT, cold exposure stimulates thermogenesis by increasing the mitochondrial content and metabolic rate. BAT and the increased metabolic rate result in a lean phenotype. This protocol provides an outstanding opportunity to study the molecular mechanisms that control adipose mass.

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

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

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

Figure 1: Postmortem dissection of iWAT and iBAT depots and low temperature–induced activation in BAT.
Figure 2: Increase of prohibitin-positive mitochondria in adipocytes by cold exposure.
Figure 3: Increase of vascular density and nonshivering thermogenesis in response to cold exposure.

Similar content being viewed by others

References

  1. Cao, Y. Angiogenesis modulates adipogenesis and obesity. J. Clin. Invest. 117, 2362–2368 (2007).

    Article  CAS  Google Scholar 

  2. Cao, Y. Adipose tissue angiogenesis as a therapeutic target for obesity and metabolic diseases. Nat. Rev. Drug Discov. 9, 107–115 (2010).

    Article  CAS  Google Scholar 

  3. Tisdale, M.J. Cachexia in cancer patients. Nat. Rev. Cancer 2, 862–871 (2002).

    Article  CAS  Google Scholar 

  4. Petrovic, N. et al. Chronic peroxisome proliferator-activated receptor γ (PPAR-γ) activation of epididymally derived white adipocyte cultures reveals a population of thermogenically competent, UCP1-containing adipocytes molecularly distinct from classic brown adipocytes. J. Biol. Chem. 285, 7153–7164 (2010).

    Article  CAS  Google Scholar 

  5. Xue, Y. et al. Hypoxia-independent angiogenesis in adipose tissues during cold acclimation. Cell Metab. 9, 99–109 (2009).

    Article  CAS  Google Scholar 

  6. Nedergaard, J. & Cannon, B. The changed metabolic world with human brown adipose tissue: therapeutic visions. Cell Metab. 11, 268–272 (2010).

    Article  CAS  Google Scholar 

  7. Feldmann, H.M., Golozoubova, V., Cannon, B. & Nedergaard, J. UCP1 ablation induces obesity and abolishes diet-induced thermogenesis in mice exempt from thermal stress by living at thermoneutrality. Cell Metab. 9, 203–209 (2009).

    Article  CAS  Google Scholar 

  8. Trayhurn, P. & Wood, I.S. Adipokines: inflammation and the pleiotropic role of white adipose tissue. Br. J. Nutr. 92, 347–355 (2004).

    Article  CAS  Google Scholar 

  9. Lowell, B.B. et al. Development of obesity in transgenic mice after genetic ablation of brown adipose tissue. Nature 366, 740–742 (1993).

    Article  CAS  Google Scholar 

  10. Xue, Y. et al. FOXC2 controls Ang-2 expression and modulates angiogenesis, vascular patterning, remodeling, and functions in adipose tissue. Proc. Natl. Acad. Sci. USA 105, 10167–10172 (2008).

    Article  CAS  Google Scholar 

  11. Cederberg, A. et al. FOXC2 is a winged helix gene that counteracts obesity, hypertriglyceridemia, and diet-induced insulin resistance. Cell 106, 563–573 (2001).

    Article  CAS  Google Scholar 

  12. Furuhashi, M. et al. Adipocyte/macrophage fatty acid-binding proteins contribute to metabolic deterioration through actions in both macrophages and adipocytes in mice. J. Clin. Invest. 118, 2640–2650 (2008).

    CAS  PubMed  PubMed Central  Google Scholar 

  13. Virtanen, K.A. et al. Functional brown adipose tissue in healthy adults. N. Engl. J. Med. 360, 1518–1525 (2009).

    Article  CAS  Google Scholar 

  14. Cypess, A.M. et al. Identification and importance of brown adipose tissue in adult humans. N. Engl. J. Med. 360, 1509–1517 (2009).

    Article  CAS  Google Scholar 

  15. van Marken Lichtenbelt, W.D. et al. Cold-activated brown adipose tissue in healthy men. N. Engl. J. Med. 360, 1500–1508 (2009).

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We thank B. Cannon and J. Nedergaard at the Stockholm University for providing the animal facility for our research. The author's laboratory was supported by research grants from the Swedish Research Council, the Swedish Cancer Foundation, the Karolinska Institute Foundation, the Karolinska Institute distinguished professor award, the Torsten Soderbergs Foundation, the European Union Integrated Project of Metoxia (project no. 222741) and the European Research Council advanced grant ANGIOFAT (project no. 250021).

Author information

Author notes

  1. Sharon Lim and Jennifer Honek: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Microbiology, Tumor and Cell Biology, Karolinska Institute, Stockholm, Sweden

    Sharon Lim, Jennifer Honek, Yuan Xue, Takahiro Seki, Patrik Andersson, Xiaojuan Yang, Kayoko Hosaka & Yihai Cao

  2. Department of Medicine and Health Sciences, Linköping University, Linköping, Sweden

    Ziquan Cao & Yihai Cao

Authors
  1. Sharon Lim
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Jennifer Honek
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Yuan Xue
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Takahiro Seki
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Ziquan Cao
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Patrik Andersson
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Xiaojuan Yang
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Kayoko Hosaka
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Yihai Cao
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

Y.C. designed the study. S.L., J.H., Y.X. and T.S. performed the experiments. S.L., J.H., Y.X., T.S. and Y.C. analyzed the data. S.L., J.H. and Y.C. wrote the paper. Z.C., P.A., X.Y. and K.H. participated in developing these protocols.

Corresponding author

Correspondence to Yihai Cao.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Lim, S., Honek, J., Xue, Y. et al. Cold-induced activation of brown adipose tissue and adipose angiogenesis in mice. Nat Protoc 7, 606–615 (2012). https://doi.org/10.1038/nprot.2012.013

Download citation

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nprot.2012.013

This article is cited by

Comments

By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.

Search

Advanced search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing