| Analyses of C. elegans Fat Metabolic Pathways. | |
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MedLine Citation:
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PMID: 22226531 Owner: NLM Status: In-Data-Review |
Abstract/OtherAbstract:
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In Caenorhabdatis elegans as in other animals, fat regulation reflects the outcome of behavioral, physiological, and metabolic processes. The amenability of C. elegans to experimentation has led to utilization of this organism for elucidating the complex homeostatic mechanisms that underlie energy balance in intact organisms. The optical advantages of C. elegans further offer the possibility of studying cell biological mechanisms of fat uptake, transport, storage, and utilization, perhaps in real time. Here, we discuss the rationale as well as advantages and potential pitfalls of methods used thus far to study metabolism and fat regulation, specifically triglyceride metabolism, in C. elegans. We provide detailed methods for visualization of fat depots in fixed animals using histochemical stains and in live animals by vital dyes. Protocols are provided and discussed for chloroform-based extraction of total lipids from C. elegans homogenates used to assess total triglyceride or phospholipid content by methods such as thin-layer chromatography or used to obtain fatty acid profiles by methods such as gas chromatography/mass spectrometry. Additionally, protocols are provided for the determination of rates of intestinal fatty acid uptake and fatty acid breakdown by β-oxidation. Finally, we discuss methods for determining rates of de novo fat synthesis and Raman scattering approaches that have recently been employed to investigate C. elegans lipids without reliance on invasive techniques. As the C. elegans fat field is relatively new, we anticipate that the indicated methods will likely be improved upon and expanded as additional researchers enter this field. |
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Authors:
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Alexandre Guimarães de Almeida Barros; Jason Liu; George A Lemieux; Brendan C Mullaney; Kaveh Ashrafi |
Publication Detail:
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Type: Journal Article |
Journal Detail:
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Title: Methods in cell biology Volume: 107 ISSN: 0091-679X ISO Abbreviation: Methods Cell Biol. Publication Date: 2012 |
Date Detail:
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Created Date: 2012-01-09 Completed Date: - Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 0373334 Medline TA: Methods Cell Biol Country: United States |
Other Details:
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Languages: eng Pagination: 383-407 Citation Subset: IM |
Copyright Information:
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Copyright © 2012 Elsevier Inc. All rights reserved. |
Affiliation:
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Laboratório de Neurociência, INCT de Medicina Molecular, Faculdade de Medicina da UFMG, Belo Horizonte, Minas Gerais, Bolsista CNPq-Brazil. |
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From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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