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Diabetes. 2017 Feb;66(2):407-417. doi: 10.2337/db16-0343. Epub 2016 Nov 8.

Visualization and Quantification of Browning Using a Ucp1-2A-Luciferase Knock-in Mouse Model.

Mao L1, Nie B2, Nie T1, Hui X3, Gao X4, Lin X5, Liu X6, Xu Y1, Tang X1, Yuan R1, Li K1, Li P1, Ding K1, Wang Y1,3, Xu A1,3, Fei J7, Han W8, Liu P4, Madsen L9,10,11, Kristiansen K10,11, Zhou Z12, Ding S13, Wu D14,15.

Author information

1
CAS Key Laboratory of Regenerative Biology, Joint School of Life Sciences, Guangzhou Medical University, and Guangzhou Institute of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou, China.
2
Gladstone Institute of Cardiovascular Disease, Department of Pharmaceutical Chemistry, University of California, San Francisco, San Francisco, CA.
3
Department of Medicine, The University of Hong Kong, Hong Kong.
4
Wellcome Sanger Institute, Cambridge, U.K.
5
Research & Development Center, Infinitus (China) Company Ltd., Guangzhou, China.
6
Shenzhen Institutes of Advanced Technology, Chinese Academy of Sciences, Shenzhen, China.
7
Shanghai Nan Fang Model Organism Research Center, Shanghai, China.
8
Singapore Bioimaging Consortium and Institute of Molecular and Cell Biology, Agency for Science, Technology and Research, Singapore.
9
National Institute of Nutrition and Seafood Research, Bergen, Norway.
10
Laboratory of Genomics and Molecular Biomedicine, Department of Biology, University of Copenhagen, Copenhagen, Denmark.
11
Beijing Genomics Institute-Shenzhen, Shenzhen, China.
12
Diabetes Center, The Second Xiangya Hospital, Institute of Metabolism and Endocrinology, Central South University, Changsha, China.
13
Gladstone Institute of Cardiovascular Disease, Department of Pharmaceutical Chemistry, University of California, San Francisco, San Francisco, CA wu_donghai@gibh.ac.cn sheng.ding@gladstone.ucsf.edu.
14
CAS Key Laboratory of Regenerative Biology, Joint School of Life Sciences, Guangzhou Medical University, and Guangzhou Institute of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou, China wu_donghai@gibh.ac.cn sheng.ding@gladstone.ucsf.edu.
15
Joint School of Biological Sciences, Guangzhou Institute of Biomedicine and Health, Guangzhou Medical University, Guangzhou, China.

Abstract

Both mammals and adult humans possess classic brown adipocytes and beige adipocytes, and the amount and activity of these adipocytes are considered key factors in combating obesity and its associated metabolic diseases. Uncoupling protein 1 (Ucp1) is the functional marker of both brown and beige adipocytes. To facilitate a reliable, easy, and sensitive measurement of Ucp1 expression both in vivo and in vitro, we generated a Ucp1-2A-luciferase knock-in mouse by deleting the stop codon for the mouse Ucp1 gene and replacing it with a 2A peptide. This peptide was followed by the luciferase coding sequence to recapitulate the expression of the Ucp1 gene at the transcriptional and translational levels. With this mouse, we discovered a cold-sensitive brown/beige adipose depot underneath the skin of the ears, which we named uBAT. Because of the sensitivity and high dynamic range of luciferase activity, the Ucp1-2A-luciferase mouse is useful for both in vitro quantitative determination and in vivo visualization of nonshivering thermogenesis. With the use of this model, we identified and characterized axitinib, an oral small-molecule tyrosine kinase inhibitor, as an effective browning agent.

PMID:
28108609
DOI:
10.2337/db16-0343
[Indexed for MEDLINE]
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