Document Detail

Analysis of mitochondrial function in situ in permeabilized muscle fibers, tissues and cells.
MedLine Citation:
PMID:  18536644     Owner:  NLM     Status:  MEDLINE    
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.
Andrey V Kuznetsov; Vladimir Veksler; Frank N Gellerich; Valdur Saks; Raimund Margreiter; Wolfram S Kunz
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Nature protocols     Volume:  3     ISSN:  1750-2799     ISO Abbreviation:  Nat Protoc     Publication Date:  2008  
Date Detail:
Created Date:  2008-06-09     Completed Date:  2008-09-03     Revised Date:  2008-11-21    
Medline Journal Info:
Nlm Unique ID:  101284307     Medline TA:  Nat Protoc     Country:  England    
Other Details:
Languages:  eng     Pagination:  965-76     Citation Subset:  IM    
Daniel Swarovski Research Laboratory, Department of General and Transplant Surgery, Innsbruck Medical University, Innsbruck A6020, Austria.
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MeSH Terms
Adenosine Diphosphate / metabolism
Adenosine Triphosphatases / metabolism
Cell Membrane Permeability
Electron Transport
Mitochondria / metabolism
Mitochondria, Muscle / metabolism*
Muscle Fibers, Skeletal / metabolism*
Oxidative Phosphorylation
Reg. No./Substance:
58-64-0/Adenosine Diphosphate; EC 3.6.1.-/Adenosine Triphosphatases

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