Analysis of mitochondrial function in situ in permeabilized muscle fibers, tissues and cells

作者：Kuznetsov Andrey V^*; Veksler Vladimir; Gellerich Frank N; Saks Valdur; Margreiter Raimund; Kunz Wolfram S

来源：Nature Protocols, 2008, 3(6): 965-976.

DOI：10.1038/nprot.2008.61

Analysis of mitochondrial function is central to the study of intracellular energy metabolism, mechanisms of cell death and pathophysiology of a variety of human diseases, including myopathies, neurodegenerative diseases and cancer. However, important properties of mitochondria differ in vivo and in vitro. Here, we describe a protocol for the analysis of functional mitochondria in situ, without the isolation of organelles, in selectively permeabilized cells or muscle fibers using digitonin or saponin. A specially designed substrate/inhibitor titration approach allows the step-by-step analysis of several mitochondrial complexes. This protocol allows the detailed characterization of functional mitochondria in their normal intracellular position and assembly, preserving essential interactions with other organelles. As only a small amount of tissue is required for analysis, the protocol can be used in diagnostic settings in clinical studies. The permeabilization procedure and specific titration analysis can be completed in 2 h.

• 出版日期2008