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Reliable Drosophila body fat quantification by a coupled colorimetric assay.

PloS one (2011-09-21)
Anja Hildebrandt, Iris Bickmeyer, Ronald P Kühnlein
ABSTRACT

Factors and mechanisms controlling lipometabolism homeostasis share a remarkable evolutionary conservation between humans and Drosophila flies. Accordingly, the Drosophila model has been successfully used to understand the pathophysiology of human metabolic diseases such as obesity. Body fat stores in species as different as humans and flies consist of neutral lipids, mainly triacylglycerols. Changes in body fat storage are a diagnostic phenotype of lipometabolism imbalances of genetic or environmental origin. Various methods have been developed to quantify Drosophila body fat storage. The most widely used method adopts a commercial coupled colorimetric assay designed for human serum triacylglycerol quantification, which is based on glycerol content determination after enzymatic conversion of glycerides into glycerol. The coupled colorimetric assay is compatible with large-scale genetic screen approaches and has been successfully applied to characterize central regulators of Drosophila lipometabolism. Recently, the applicability of the coupled colorimetric assay for Drosophila storage fat quantification has been questioned in principle. Here we compare the performance of the coupled colorimetric assay on Drosophila samples with thin layer chromatography, the "gold standard" in storage lipid analysis. Our data show that the presented variant of the coupled colorimetric assay reliably discriminates between lean and fat flies and allows robust, quick and cost-effective quantification of Drosophila body fat stores.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Triglyceride Reagent, used for quantitative enzymatic determination of triglyerides
Sigma-Aldrich
Glyceryl trioleate, ≥99%
Sigma-Aldrich
Glycerol Standard Solution, 2.5 mg/ml equivalent triolein concentration
Sigma-Aldrich
Free Glycerol Reagent, used for quantitative enzymatic determination of glycerol