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Measuring O2 Consumption in Drosophila Melanogaster using Coulometric Microrespirometry

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JOURNAL OF VISUALIZED EXPERIMENTS
DOI: 10.3791/65379

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Coulometric microrespirometry is a simple and cost-effective method for measuring the O2 consumption of small organisms. It utilizes a sealed chamber where O2 is consumed and CO2 is removed. The decrease in pressure triggers electrolytic O2 production, which is measured by the amount of charge used.
Coulometric microrespirometry is a straightforward, inexpensive method for measuring the O2 consumption of small organisms while maintaining a stable environment. A coulometric microrespirometer consists of an airtight chamber in which O2 is consumed and the CO2 produced by the organism is removed by an absorbent medium. The resulting pressure decrease triggers electrolytic O2 production, and the amount of O2 produced is measured by recording the amount of charge used to generate it. In the present study, the method has been adapted to Drosophila melanogaster tested in small groups, with the sensitivity of the apparatus and the environmental conditions optimized for high stability. The amount of O2 consumed by wildtype flies in this apparatus is consistent with that measured by previous studies. Mass-specific O2 consumption by CASK mutants, which are smaller and known to be less active, was not different from congenic controls. However, the small size of CASK mutants resulted in a significant reduction in O2 consumption on a per-fly basis. Therefore, the microrespirometer is capable of measuring O2 consumption in D. melanogaster, can distinguish modest differences between genotypes, and adds a versatile tool for measuring metabolic rates.

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