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Measurement of Carbon Dioxide Production from Radiolabeled Substrates in Drosophila melanogaster

The power of Drosophila genetics is increasingly being applied to questions of hormone signaling and metabolism and to the development of models of human disease in this organism. Sensitive methods for measurements of parameters such as metabolic rates are needed to drive the understanding of physio...

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Autor principal: Bland, Michelle L.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MyJove Corporation 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4993251/
https://www.ncbi.nlm.nih.gov/pubmed/27404635
http://dx.doi.org/10.3791/54045
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author Bland, Michelle L.
author_facet Bland, Michelle L.
author_sort Bland, Michelle L.
collection PubMed
description The power of Drosophila genetics is increasingly being applied to questions of hormone signaling and metabolism and to the development of models of human disease in this organism. Sensitive methods for measurements of parameters such as metabolic rates are needed to drive the understanding of physiology and disease in small animals such as the fruit fly. The method described here assesses fuel oxidation in small numbers of adult flies fed food containing trace amounts of (14)C-labeled substrates such as glucose or fatty acid. After the feeding period and any additional experimental manipulations, flies are transferred to short tubes capped with mesh, which are then placed in glass vials containing KOH-saturated filter paper that traps exhaled, radiolabeled CO(2) generated from oxidation of radiolabeled substrates as potassium bicarbonate, KHCO(3). This radiolabeled bicarbonate is measured by scintillation counting. This is a quantitative, reproducible, and simple approach for the study of fuel oxidation. The use of radiolabeled glucose, fatty acids, or amino acids allows determination of the contribution of these different fuel sources to energy metabolism under different conditions such as feeding and fasting and in different genetic backgrounds. This complements other approaches used to measure in vivo energy metabolism and should further the understanding of metabolic regulation.
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spelling pubmed-49932512016-09-20 Measurement of Carbon Dioxide Production from Radiolabeled Substrates in Drosophila melanogaster Bland, Michelle L. J Vis Exp Cellular Biology The power of Drosophila genetics is increasingly being applied to questions of hormone signaling and metabolism and to the development of models of human disease in this organism. Sensitive methods for measurements of parameters such as metabolic rates are needed to drive the understanding of physiology and disease in small animals such as the fruit fly. The method described here assesses fuel oxidation in small numbers of adult flies fed food containing trace amounts of (14)C-labeled substrates such as glucose or fatty acid. After the feeding period and any additional experimental manipulations, flies are transferred to short tubes capped with mesh, which are then placed in glass vials containing KOH-saturated filter paper that traps exhaled, radiolabeled CO(2) generated from oxidation of radiolabeled substrates as potassium bicarbonate, KHCO(3). This radiolabeled bicarbonate is measured by scintillation counting. This is a quantitative, reproducible, and simple approach for the study of fuel oxidation. The use of radiolabeled glucose, fatty acids, or amino acids allows determination of the contribution of these different fuel sources to energy metabolism under different conditions such as feeding and fasting and in different genetic backgrounds. This complements other approaches used to measure in vivo energy metabolism and should further the understanding of metabolic regulation. MyJove Corporation 2016-06-27 /pmc/articles/PMC4993251/ /pubmed/27404635 http://dx.doi.org/10.3791/54045 Text en Copyright © 2016, Journal of Visualized Experiments http://creativecommons.org/licenses/by-nc-nd/3.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs 3.0 Unported License. To view a copy of this license, visithttp://creativecommons.org/licenses/by-nc-nd/3.0/
spellingShingle Cellular Biology
Bland, Michelle L.
Measurement of Carbon Dioxide Production from Radiolabeled Substrates in Drosophila melanogaster
title Measurement of Carbon Dioxide Production from Radiolabeled Substrates in Drosophila melanogaster
title_full Measurement of Carbon Dioxide Production from Radiolabeled Substrates in Drosophila melanogaster
title_fullStr Measurement of Carbon Dioxide Production from Radiolabeled Substrates in Drosophila melanogaster
title_full_unstemmed Measurement of Carbon Dioxide Production from Radiolabeled Substrates in Drosophila melanogaster
title_short Measurement of Carbon Dioxide Production from Radiolabeled Substrates in Drosophila melanogaster
title_sort measurement of carbon dioxide production from radiolabeled substrates in drosophila melanogaster
topic Cellular Biology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4993251/
https://www.ncbi.nlm.nih.gov/pubmed/27404635
http://dx.doi.org/10.3791/54045
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