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1. chinaXiv:201711.02415 [pdf]

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

Mao, LF; Nie, BM; Nie, T; Hui, XY; Gao, XF; Lin, XL; Liu, X; Xu, Y; Tang, XF; Yuan, R; Li, K; Li, P; Ding, K; Wang, Y; Xu, AM; Fei, J; Han, WP; Liu, PT; Madsen, L; Kristiansen, K
Subjects: Biology >> Ecology

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.

submitted time 2017-11-17 Hits1048Downloads617 Comment 0

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