Fluorescent and Luminescent Methods to Detect Lipolysis.

ABSTRACT

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.