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Methods DOI:10.1016/j.ymeth.2014.04.017

High- and low-throughput scoring of fat mass and body fat distribution in C. elegans.

Publication TypeJournal Article
Year of Publication2014
AuthorsWählby, C, Conery, ALee, Bray, M-A, Kamentsky, L, Larkins-Ford, J, Sokolnicki, KL, Veneskey, M, Michaels, K, Carpenter, AE, O'Rourke, EJ
JournalMethods
Volume68
Issue3
Pages492-9
Date Published2014 Aug 01
ISSN1095-9130
KeywordsAnimals, Body Fat Distribution, Caenorhabditis elegans, Genome, High-Throughput Screening Assays, Lipid Metabolism, Models, Animal, Obesity, Phenotype
Abstract

Fat accumulation is a complex phenotype affected by factors such as neuroendocrine signaling, feeding, activity, and reproductive output. Accordingly, the most informative screens for genes and compounds affecting fat accumulation would be those carried out in whole living animals. Caenorhabditis elegans is a well-established and effective model organism, especially for biological processes that involve organ systems and multicellular interactions, such as metabolism. Every cell in the transparent body of C. elegans is visible under a light microscope. Consequently, an accessible and reliable method to visualize worm lipid-droplet fat depots would make C. elegans the only metazoan in which genes affecting not only fat mass but also body fat distribution could be assessed at a genome-wide scale. Here we present a radical improvement in oil red O worm staining together with high-throughput image-based phenotyping. The three-step sample preparation method is robust, formaldehyde-free, and inexpensive, and requires only 15min of hands-on time to process a 96-well plate. Together with our free and user-friendly automated image analysis package, this method enables C. elegans sample preparation and phenotype scoring at a scale that is compatible with genome-wide screens. Thus we present a feasible approach to small-scale phenotyping and large-scale screening for genetic and/or chemical perturbations that lead to alterations in fat quantity and distribution in whole animals.

URLhttp://linkinghub.elsevier.com/retrieve/pii/S1046-2023(14)00170-4
DOI10.1016/j.ymeth.2014.04.017
Pubmed

http://www.ncbi.nlm.nih.gov/pubmed/24784529?dopt=Abstract

Alternate JournalMethods
PubMed ID24784529
PubMed Central IDPMC4112171
Grant ListR00 DK087928 / DK / NIDDK NIH HHS / United States
R01 GM089652 / GM / NIGMS NIH HHS / United States
R01 GM095672 / GM / NIGMS NIH HHS / United States