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Colorimetric Measurement of Triglycerides Cannot Provide an Accurate Measure of Stored Fat Content in Drosophila

Al-Anzi, Bader and Zinn, Kai (2010) Colorimetric Measurement of Triglycerides Cannot Provide an Accurate Measure of Stored Fat Content in Drosophila. PLoS ONE, 5 (8). Art. No. e12353. ISSN 1932-6203. PMCID PMC2927426. https://resolver.caltech.edu/CaltechAUTHORS:20100913-132751531

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Abstract

Drosophila melanogaster has recently emerged as a useful model system in which to study the genetic basis of regulation of fat storage. One of the most frequently used methods for evaluating the levels of stored fat (triglycerides) in flies is a coupled colorimetric assay available as a kit from several manufacturers. This is an aqueous-based enzymatic assay that is normally used for measurement of mammalian serum triglycerides, which are present in soluble lipoprotein complexes. In this short communication, we show that coupled colorimetric assay kits cannot accurately measure stored triglycerides in Drosophila. First, they fail to give accurate readings when tested on insoluble triglyceride mixtures with compositions like that of stored fat, or on fat extracted from flies with organic solvents. This is probably due to an inability of the lipase used in the kits to efficiently cleave off the glycerol head group from fat molecules in insoluble samples. Second, the measured final products of the kits are quinoneimines, which absorb visible light in the same wavelength range as Drosophila eye pigments. Thus, when extracts from crushed flies are assayed, much of the measured signal is actually due to eye pigments. Finally, the lipoprotein lipases used in colorimetric assays also cleave non-fat glycerides. The glycerol backbones liberated from all classes of glycerides are measured through the remaining reactions in the assay. As a consequence, when these assay kits are used to evaluate tissue extracts, the observed signal actually represents the amount of free glycerols together with all types of glycerides. For these reasons, findings obtained through use of coupled colorimetric assays on Drosophila samples must be interpreted with caution. We also show here that using thin-layer chromatography to measure stored triglycerides in flies eliminates all of these problems.


Item Type:Article
Related URLs:
URLURL TypeDescription
http://dx.doi.org/10.1371/journal.pone.0012353 DOIArticle
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2927426/PubMed CentralArticle
ORCID:
AuthorORCID
Zinn, Kai0000-0002-6706-5605
Additional Information:© 2010 Al-Anzi, Zinn. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Received May 21, 2010; Accepted July 18, 2010; Published August 24, 2010. Editor: Andreas Bergmann, University of Texas MD Anderson Cancer Center, United States of America. Funding: NIH RO1 grants DK0170154, NS28182. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. Author Contributions: Conceived and designed the experiments: BFAA. Performed the experiments: BFAA. Analyzed the data: BFAA KZ. Contributed reagents/materials/analysis tools: BFAA. Wrote the paper: KZ.
Funders:
Funding AgencyGrant Number
NIHRO1 DK0170154
NIHRO1 NS28182
Issue or Number:8
PubMed Central ID:PMC2927426
Record Number:CaltechAUTHORS:20100913-132751531
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20100913-132751531
Official Citation:Al-Anzi B, Zinn K (2010) Colorimetric Measurement of Triglycerides Cannot Provide an Accurate Measure of Stored Fat Content in Drosophila. PLoS ONE 5(8): e12353. doi:10.1371/journal.pone.0012353
Usage Policy:This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
ID Code:19910
Collection:CaltechAUTHORS
Deposited By: Tony Diaz
Deposited On:16 Sep 2010 21:38
Last Modified:03 Oct 2019 02:03

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