通过细胞或分离细胞器中的呼吸测定法评估线粒体生物能量学

Assessing Mitochondrial Bioenergetics by Respirometry in Cells or Isolated Organelles.

作者信息

Vial Guillaume, Guigas Bruno

机构信息

INSERM U1042, Laboratoire Hypoxie-Physiopathologies cardiovasculaires et respiratoires HP2, Faculté de médecine et de pharmacie, Domaine de la merci, La Tronche, France.

Université Grenoble-Alpes, Laboratoire Hypoxie-Physiopathologies cardiovasculaires et respiratoires HP2, Faculté de médecine et de pharmacie, Domaine de la merci, La Tronche, France.

出版信息

Methods Mol Biol. 2018;1732:273-287. doi: 10.1007/978-1-4939-7598-3_18.

DOI:10.1007/978-1-4939-7598-3_18

PMID:29480482

Abstract

Mitochondrial oxidative phosphorylation is central for generating ATP and maintaining energy homeostasis in most eukaryotic cells. The ex vivo measurement of mitochondrial oxygen consumption rates in intact cells or isolated organelles is a valuable approach to assess mitochondrial bioenergetics in various experimental conditions. In this chapter, we describe several step-by-step protocols for measuring mitochondrial respiration in intact cells, permeabilized cells (in situ mitochondria), and isolated organelles using both Clark-type polarographic oxygen electrode devices and the newly developed oxygen-sensing fluorophore-based Seahorse technology.

摘要

线粒体氧化磷酸化对于大多数真核细胞中ATP的产生和能量稳态的维持至关重要。在完整细胞或分离的细胞器中对线粒体氧消耗率进行离体测量，是在各种实验条件下评估线粒体生物能量学的一种有价值的方法。在本章中，我们描述了几种逐步方案，用于使用Clark型极谱氧电极装置和新开发的基于氧敏感荧光团的海马技术，测量完整细胞、透化细胞（原位线粒体）和分离的细胞器中的线粒体呼吸。