Department of Internal Medicine, Hypertension and Vascular Research Division, Henry Ford Hospital, Detroit, MI, USA.
Department of Physiology, Wayne State University School of Medicine, Detroit, MI, USA.
Methods Mol Biol. 2022;2448:97-106. doi: 10.1007/978-1-0716-2087-8_6.
Intracellular lipolysis, the hydrolysis of stored triacylglycerol to fatty acids and glycerol, is a core metabolic function of brown and white adipocytes. In brown adipocytes, mobilized fatty acids directly activate uncoupling protein 1, provide fuel for heat generation, and ligands of nuclear receptors that expand the thermogenic gene expression program. Lipolysis in white adipocytes mobilizes lipid energy for systemic use, including both shivering and non-shivering thermogenesis. In addition, most metabolic tissues, including muscle and liver, have the ability to store triacylglycerol and release fatty acids; thus, there is a general interest in measuring lipolysis in a wide array of cell types. Here we describe detailed protocols for the enzymatic detection of cellular fatty acid and glycerol efflux via fluorescent and colorimetric means, respectively. In addition, we also describe a genetically encoded luminescent detection system for intracellular fatty acids that is amenable to high-throughput analysis.
细胞内脂解作用，即将储存的三酰甘油水解为脂肪酸和甘油，是棕色和白色脂肪细胞的核心代谢功能。在棕色脂肪细胞中，动员的脂肪酸直接激活解偶联蛋白 1，为产热提供燃料，并为核受体配体提供燃料，这些配体可扩展生热基因表达程序。白色脂肪细胞中的脂解作用可动员脂质能量供全身使用，包括颤抖和非颤抖产热。此外，包括肌肉和肝脏在内的大多数代谢组织都有储存三酰甘油和释放脂肪酸的能力；因此，人们普遍有兴趣在广泛的细胞类型中测量脂解作用。在这里，我们描述了通过荧光和比色法分别检测细胞脂肪酸和甘油外排的详细酶促测定方案。此外，我们还描述了一种可用于高通量分析的用于细胞内脂肪酸的遗传编码发光检测系统。
