Dubikovskaya Elena, Chudnovskiy Rostislav, Karateev Grigory, Park Hyo Min, Stahl Andreas
Department of Chemistry, École Polytechnique Fédérale de Lausanne, Lausanne, Switzerland.
Department of Nutritional Science and Toxicology, UC Berkeley, Berkeley, California, USA.
Methods Enzymol. 2014;538:107-34. doi: 10.1016/B978-0-12-800280-3.00007-4.
The ability of white and brown adipose tissue to efficiently take up long-chain fatty acids is key to their physiological functions in energy storage and thermogenesis, respectively. Several approaches have been taken to determine uptake rates by cultured cells and primary adipocytes including radio- and fluorescently labeled fatty acids. In addition, the recent description of activatable bioluminescent fatty acids has opened the possibility for expanding these in vitro approaches to real-time monitoring of fatty acid uptake kinetics by adipose depots in vivo. Here, we will describe some of the most useful experimental paradigms to quantitatively determine long-chain fatty acid uptake by adipocytes in vitro and provide the reader with detailed instruction on how bioluminescent probes for in vivo imaging can be synthesized and used in living mice.
白色和棕色脂肪组织有效摄取长链脂肪酸的能力,分别是它们在能量储存和产热方面生理功能的关键。已经采用了几种方法来测定培养细胞和原代脂肪细胞的摄取率,包括放射性和荧光标记的脂肪酸。此外,可激活生物发光脂肪酸的最新描述,为将这些体外方法扩展到体内脂肪库对脂肪酸摄取动力学的实时监测开辟了可能性。在这里,我们将描述一些最有用的实验范式,以定量测定体外脂肪细胞对长链脂肪酸的摄取,并为读者提供有关如何合成用于体内成像的生物发光探针以及如何在活体小鼠中使用的详细说明。