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.

  • Technical Report
  • Published:

An OLTAM system for analysis of brown/beige fat thermogenic activity

International Journal of Obesity volume 42, pages 939–945 (2018)Cite this article

Subjects

Abstract

Background/Objectives:

Thermogenic fat is present in humans and emerging evidence indicates that increasing the content and activity of these adipocytes may lead to weight loss and improved metabolic health. Multiple reporter systems have been developed to assay thermogenic fat activity based on the transcriptional and translational activation of Ucp1, the key molecule that mediates nonshivering thermogenesis. Our study aims to develop a much-needed tool to monitor thermogenic fat activity through a mechanism independent of Ucp1 regulation, therefore effectively assaying not only canonical β-adrenergic activation but also various non-UCP1-mediated thermogenic pathways that have been increasingly appreciated.

Methods:

We detected increased luciferase activity upon thermogenic activation in interscapular brown and inguinal subcutaneous fat in ODD-Luc mice, a hypoxia reporter mouse model. We then developed an OLTAM ( O DD- L uc based T hermogenic A ctivity M easurement) system to assay thermogenic fat cell activity.

Results:

In both primary murine and human adipocytes and an immortalized adipose cell line that were transduced with the OLTAM system, luciferase activity can be readily measured and visualized by bioluminescence imaging in response to a variety of stimuli, including UCP1-independent thermogenic signaling. This system can offer a convenient method to assay thermogenic activity for both basic and translational research.

Conclusions:

The OLTAM system offers a convenient way to measure the activation of thermogenic fat and presents opportunities to discover novel signaling pathways and unknown compounds targeting metabolically active adipocytes to counteract human obesity.

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
Figure 2
Figure 3
Figure 4

Similar content being viewed by others

References

  1. Kajimura S, Spiegelman BM, Seale P . Brown and beige fat: physiological roles beyond heat generation. Cell Metab 2015; 22: 546–559.

    Article  CAS  Google Scholar 

  2. Hanssen MJ, Hoeks J, Brans B, van der Lans AA, Schaart G, van den Driessche JJ et al. Short-term cold acclimation improves insulin sensitivity in patients with type 2 diabetes mellitus. Nat med 2015; 21: 863–865.

    Article  CAS  Google Scholar 

  3. Sidossis L, Kajimura S . Brown and beige fat in humans: thermogenic adipocytes that control energy and glucose homeostasis. J clin invest 2015; 125: 478–486.

    Article  Google Scholar 

  4. Galmozzi A, Sonne SB, Altshuler-Keylin S, Hasegawa Y, Shinoda K, Luijten IH et al. ThermoMouse: an in vivo model to identify modulators of UCP1 expression in brown adipose tissue. Cell rep 2014; 9: 1584–1593.

    Article  CAS  Google Scholar 

  5. Mao L, Nie B, Nie T, Hui X, Gao X, Lin X et al. Visualization and quantification of browning using a Ucp1-2A-luciferase knock-in mouse model. Diabetes 2017; 66: 407–417.

    Article  CAS  Google Scholar 

  6. Kazak L, Chouchani ET, Jedrychowski MP, Erickson BK, Shinoda K, Cohen P et al. A creatine-driven substrate cycle enhances energy expenditure and thermogenesis in beige fat. Cell 2015; 163: 643–655.

    Article  CAS  Google Scholar 

  7. Shabalina IG, Kramarova TV, Nedergaard J, Cannon B . Carboxyatractyloside effects on brown-fat mitochondria imply that the adenine nucleotide translocator isoforms ANT1 and ANT2 may be responsible for basal and fatty-acid-induced uncoupling respectively. Biochem J 2006; 399: 405–414.

    Article  CAS  Google Scholar 

  8. Long JZ, Svensson KJ, Bateman LA, Lin H, Kamenecka T, Lokurkar IA et al. The secreted enzyme PM20D1 regulates lipidated amino acid uncouplers of mitochondria. Cell 2016; 166: 424–435.

    Article  CAS  Google Scholar 

  9. Foster DO, Frydman ML . Nonshivering thermogenesis in the rat. II. Measurements of blood flow with microspheres point to brown adipose tissue as the dominant site of the calorigenesis induced by noradrenaline. Can J Physiol Pharmacol 1978; 56: 110–122.

    Article  CAS  Google Scholar 

  10. Xue Y, Petrovic N, Cao R, Larsson O, Lim S, Chen S et al. Hypoxia-independent angiogenesis in adipose tissues during cold acclimation. Cell Metab 2009; 9: 99–109.

    Article  CAS  Google Scholar 

  11. Safran M, Kim WY, O'Connell F, Flippin L, Gunzler V, Horner JW et al. Mouse model for noninvasive imaging of HIF prolyl hydroxylase activity: assessment of an oral agent that stimulates erythropoietin production. Proc Natl Acad Sci USA 2006; 103: 105–110.

    Article  CAS  Google Scholar 

  12. Cannon B, Nedergaard J . Brown adipose tissue: function and physiological significance. Physiol Rev 2004; 84: 277–359.

    Article  CAS  Google Scholar 

  13. Wu J, Bostrom P, Sparks LM, Ye L, Choi JH, Giang AH et al. Beige adipocytes are a distinct type of thermogenic fat cell in mouse and human. Cell 2012; 150: 366–376.

    Article  CAS  Google Scholar 

  14. Handschin C, Spiegelman BM . Peroxisome proliferator-activated receptor gamma coactivator 1 coactivators, energy homeostasis, and metabolism. Endocr Rev 2006; 27: 728–735.

    Article  CAS  Google Scholar 

  15. Wilson-Fritch L, Burkart A, Bell G, Mendelson K, Leszyk J, Nicoloro S et al. Mitochondrial biogenesis and remodeling during adipogenesis and in response to the insulin sensitizer rosiglitazone. Mol cell biol 2003; 23: 1085–1094.

    Article  CAS  Google Scholar 

  16. Ohno H, Shinoda K, Spiegelman BM, Kajimura S . PPARgamma agonists induce a white-to-brown fat conversion through stabilization of PRDM16 protein. Cell Metab 2012; 15: 395–404.

    Article  CAS  Google Scholar 

  17. Kajimura S, Seale P, Kubota K, Lunsford E, Frangioni JV, Gygi SP et al. Initiation of myoblast to brown fat switch by a PRDM16-C/EBP-beta transcriptional complex. Nature 2009; 460: 1154–1158.

    Article  CAS  Google Scholar 

  18. Elsen M, Raschke S, Tennagels N, Schwahn U, Jelenik T, Roden M et al. BMP4 and BMP7 induce the white-to-brown transition of primary human adipose stem cells. Am J Physiol Cell Physiol 2014; 306: C431–C440.

    Article  CAS  Google Scholar 

  19. Bartesaghi S, Hallen S, Huang L, Svensson PA, Momo RA, Wallin S et al. Thermogenic activity of UCP1 in human white fat-derived beige adipocytes. Mol Endocrinol 2015; 29: 130–139.

    Article  Google Scholar 

  20. Odegaard JI, Lee MW, Sogawa Y, Bertholet AM, Locksley RM, Weinberg DE et al. Perinatal licensing of thermogenesis by IL-33 and ST2. Cell 2016; 166: 841–854.

    Article  CAS  Google Scholar 

  21. Takahashi A, Adachi S, Morita M, Tokumasu M, Natsume T, Suzuki T et al. Post-transcriptional stabilization of Ucp1 mRNA protects mice from diet-induced obesity. Cell Rep 2015; 13: 2756–2767.

    Article  CAS  Google Scholar 

  22. Chouchani ET, Kazak L, Jedrychowski MP, Lu GZ, Erickson BK, Szpyt J et al. Mitochondrial ROS regulate thermogenic energy expenditure and sulfenylation of UCP1. Nature 2016; 532: 112–116.

    Article  CAS  Google Scholar 

  23. Ouellet V, Labbe SM, Blondin DP, Phoenix S, Guerin B, Haman F et al. Brown adipose tissue oxidative metabolism contributes to energy expenditure during acute cold exposure in humans. J Clin Invest 2012; 122: 545–552.

    Article  Google Scholar 

  24. Muzik O, Mangner TJ, Leonard WR, Kumar A, Janisse J, Granneman JG . 15O PET measurement of blood flow and oxygen consumption in cold-activated human brown fat. J Nucl Med 2013; 54: 523–531.

    Article  CAS  Google Scholar 

  25. Krohn KA, Link JM, Mason RP . Molecular imaging of hypoxia. J Nucl Med 2008; 49 (Suppl 2): 129S–148S.

    Article  CAS  Google Scholar 

Download references

Acknowledgements

This research was supported by the Edward Mallinckrodt Jr. Foundation Mallinckrodt grant (to JW), NIH Grant R01 DK107583 (to JW), NIH Grant R01 DK095201 (to YMS), NIH Grant R01 DK47918 (to MBO), NIH Grant F31DK112625 (to MPE), a postdoctoral fellowship from the American Heart Association 17POST33060001 (to D-IK), Clinical and Translational Science Award Grant UL1TR000433 (to M-JP) and NIH grant P30 DK34933 to the University of Michigan.

Author information

Authors and Affiliations

  1. Life Sciences Institute, University of Michigan, Ann Arbor, MI, USA

    D-I Kim, J Liao, M P Emont, H Jun, J D Lin & J Wu

  2. Department of Endocrinology and Metabolism, 2nd Xiangya Hospital, Central South University, Changsha, Hunan, China

    J Liao

  3. Department of Molecular & Integrative Physiology, University of Michigan, Ann Arbor, MI, USA

    M P Emont, M-J Park, S K Ramakrishnan, Y M Shah, M B Omary & J Wu

  4. Department of Cell & Developmental Biology, University of Michigan, Ann Arbor, MI, USA

    J D Lin

Authors
  1. D-I Kim
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. J Liao
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. M P Emont
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. M-J Park
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. H Jun
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. S K Ramakrishnan
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. J D Lin
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Y M Shah
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. M B Omary
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. J Wu
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to J Wu.

Ethics declarations

Competing interests

The authors declare no conflict of interest.

Additional information

Supplementary Information accompanies this paper on International Journal of Obesity website

Supplementary information

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Kim, DI., Liao, J., Emont, M. et al. An OLTAM system for analysis of brown/beige fat thermogenic activity. Int J Obes 42, 939–945 (2018). https://doi.org/10.1038/ijo.2017.308

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/ijo.2017.308

This article is cited by

Search

Advanced search

Quick links