title: A Buoyancy-based Method of Determining Fat Levels in Drosophila.
abstract: Drosophila melanogaster is a key experimental system in the study of fat regulation. Numerous techniques currently exist to measure levels of stored fat in Drosophila, but most are expensive and/or laborious and have clear limitations. Here, we present a method to quickly and cheaply determine organismal fat levels in L3 Drosophila larvae. The technique relies on the differences in density between fat and lean tissues and allows for rapid detection of fat and lean phenotypes. We have verified the accuracy of this method by comparison to body fat percentage as determined by neutral lipid extraction and gas chromatography coupled with mass spectrometry (GCMS). We furthermore outline detailed protocols for the collection and synchronization of larvae as well as relevant experimental recipes. The technique presented below overcomes the major shortcomings in the most widely used lipid quantitation methods and provides a powerful way to quickly and sensitively screen L3 larvae for fat regulation phenotypes while maintaining the integrity of the larvae. This assay has wide applications for the study of metabolism and fat regulation using Drosophila.
authors.affiliation: Department of Medicine, Division of Endocrinology, Metabolism, and Diabetes, University of Colorado Anschutz Medical Campus.
authors.forename: Kelsey E
authors.initials: KE
authors.lastname: Hazegh
authors.affiliation: Department of Medicine, Division of Endocrinology, Metabolism, and Diabetes, University of Colorado Anschutz Medical Campus; tania.reis@ucdenver.edu.
authors.forename: Tânia
authors.initials: T
authors.lastname: Reis
mesh_terms: D000818:Animals; D004331:Drosophila melanogaster / Q000378:metabolism*; D005223:Fats / Q000032:analysis* / Q000378:metabolism*; D007814:Larva
chemical_list: D005223:Fats