Raw data for the manuscript: Harmonising protocols to measure Drosophila respiratory function in mitochondrial preparations

Recent advances in fluorespirometry have allowed to measure multiple parameters of mitochondrial function; yet protocols vary widely between research groups, organisms, and tissues, sometimes without adequate justification or with assumptions that are not supported by data. This is particularly clear in Drosophila studies, a common model organism in biological research, with increasingly popularity in mitochondrial research. We first surveyed the literature on Drosophila fluorespirometry to compare and contrast approaches used in the field. Then, we showed that contrary to previous assumptions based on mammalian studies, oxygen diffusion is not limited in the permeabilised thoracic muscle of this species. We further compared the permeabilization and homogenising approaches, showing the former to be more adequate in Drosophila. Finally, we assessed the concentration-dependent effect of various commonly used fluorescent probes on mitochondrial respiration, showing that some probes strongly affect mitochondria and that careful choice of concentration and probe needs to be taken. The objective of this study is to give recommendations and work towards a harmonisation of the protocols for assessing mitochondrial function in fruit flies. This upload contains the raw data supporting these findings.

荧光光谱技术的近期进展使得测量线粒体功能的多项参数成为可能；然而，在研究小组、生物体和器官组织之间，实验方案存在广泛的差异，有时缺乏充分的论证或基于未经数据支持的假设。这一现象在果蝇研究中尤为明显，果蝇作为生物研究中的常见模式生物，在线粒体研究中的地位日益凸显。我们首先对果蝇荧光光谱文献进行了综述，以比较和对比该领域所采用的方法。随后，我们发现，与基于哺乳动物研究的先前假设相反，氧气扩散并非在渗透性胸肌中受限。我们进一步比较了渗透化和均质化方法，证明前者在果蝇中更为适宜。最后，我们评估了各种常用荧光探针对线粒体呼吸的浓度依赖性影响，表明某些探针会显著影响线粒体，且需谨慎选择浓度和探针。本研究的目的是提供建议，并致力于协调评估果蝇线粒体功能的实验方案。本上传包含支持这些发现的原生数据。