Rodriguez, Enrique; Bettinazzi, Stefano; Inwongwan, Sahutchai; Camus, Florencia; Lane, Nick; (2023) Harmonizing protocols to measure Drosophila respiratory function in mitochondrial preparations. Bioenergetics Communications (BEC) , 2023.3 pp. 1-25. 10.26124/bec:2023-0003.

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Abstract

Recent advances in fluorespirometry have enabled measurements of multiple mitochondrial parameters. However, specific protocols vary widely between research groups, organisms, and tissues, sometimes without adequate justification, or based on assumptions not supported by data. This is particularly clear in Drosophila studies, a common model organism in biomedical research, which is increasingly popular in mitochondrial studies. We first surveyed the literature on Drosophila fluorespirometry to compare and contrast approaches used in the field. Then, we showed that, in contrast to assumptions based on mammalian studies, oxygen diffusion is not limited in the permeabilized thoracic muscle of this species. We further compared permeabilization and homogenization approaches, showing that the former are better in Drosophila thoraces. Finally, we assessed the concentration-dependent effect of various commonly used fluorescent probes on mitochondrial respiration, showing that some probes strongly affect mitochondria: care needs to be taken in the choice of concentration and probe. We recommend working towards the harmonization of protocols for assessing mitochondrial function in fruit flies.

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