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