Federal University of Rio de Janeiro, Institute of Medical Biochemistry Leopoldo de Meis, Rio De Janeiro, RJ, Brazil.
Methods Mol Biol. 2021;2276:67-85. doi: 10.1007/978-1-0716-1266-8_5.
Respirometry analysis is an effective technique to assess mitochondrial physiology. Insects are valuable biochemical models to understand metabolism and human diseases. Insect flight muscle and brain have been extensively used to explore mitochondrial function due to dissection feasibility and the low sample effort to allow oxygen consumption measurements. However, adequate plasma membrane permeabilization is required for substrates/modulators to reach mitochondria. Here, we describe a new method for study of mitochondrial physiology in insect tissues based on mechanical permeabilization as a fast and reliable method that do not require the use of detergents for chemical permeabilization of plasma membrane, while preserves mitochondrial integrity.
呼吸测量分析是评估线粒体生理学的有效技术。昆虫是研究代谢和人类疾病的有价值的生化模型。昆虫飞行肌肉和大脑由于其易于解剖，并且样本需求量低，可用于测量耗氧量，因此被广泛用于探索线粒体功能。然而，为了使底物/调节剂到达线粒体，需要充分的质膜通透性。在这里，我们描述了一种基于机械通透化的昆虫组织中线粒体生理学研究的新方法，该方法快速可靠，不需要使用去污剂进行质膜的化学通透化，同时保持线粒体的完整性。
