Chair for Molecular Nutritional Medicine, TUM School of Life Sciences Weihenstephan, Technical University of Munich, Gregor-Mendel-Str. 2, 85354 Freising, Germany.
EKFZ - Else Kröner-Fresenius Center for Nutritional Medicine, Technical University of Munich, Gregor-Mendel-Str. 2, 85354 Freising, Germany.
STAR Protoc. 2020 Sep 22;1(3):100118. doi: 10.1016/j.xpro.2020.100118. eCollection 2020 Dec 18.
Studying brown and brite adipose tissue requires precise and reliable quantification of cellular thermogenesis. This protocol describes the isolation of primary murine pre-adipocytes, differentiation into thermogenic brown and brite adipocytes, and subsequent oxygen consumption analysis. Commonly applied procedures only measure basal and maximal proton leak-linked oxygen consumption but not explicitly uncoupling protein 1 (UCP1)-dependent respiration. Meaningful oxygen consumption analyses require (1) the activation of UCP1, (2) control over intracellular free-fatty-acid levels, and (3) inhibition of ATP-consuming futile cycles. For complete details on the use and execution of this protocol, please refer to Li et al. (2014, 2017, 2018) and Schweizer et al. (2018).
研究棕色和米色脂肪组织需要精确可靠地量化细胞产热。本方案描述了从小鼠原代前体脂肪细胞中分离、分化为产热棕色和米色脂肪细胞，以及随后进行耗氧分析的过程。常用的方法仅测量基础和最大质子漏相关的耗氧，但不能明确测量解偶联蛋白 1（UCP1）依赖性呼吸。要进行有意义的耗氧分析，需要（1）激活 UCP1，（2）控制细胞内游离脂肪酸水平，以及（3）抑制消耗 ATP 的无效循环。如需详细了解本方案的使用和执行情况，请参考 Li 等人（2014 年、2017 年、2018 年）和 Schweizer 等人（2018 年）的研究。
