Analyzing mitochondrial changes during apoptosis.

Roberta A Gottlieb, David J Granville

Journal: Methods (San Diego, Calif.) 2002;26(4):341-7

PMID: 12054925

Full Text on Elsevier Science View on PubMed

Abstract

Mitochondria play a central role in programmed cell death through the release of cytochrome c and other proapoptotic factors. Fluorescence microscopy is used to visualize cytochrome c translocation and loss of mitochondrial membrane potential. Flow cytometry can also be used to measure mitochondrial membrane potential. Cytochrome c content in cytosol and mitochondria can be determined by immunoblotting after subcellular fractionation or selective permeabilization with digitonin. Isolated mitochondria can be used to study the mechanism of cytochrome c release. This article summarizes some of the more widely used methods to assess mitochondrial alterations in apoptosis.

Address: Department of Molecular and Experimental Medicine, The Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, CA 92037, USA. [email protected]

DOI: 10.1016/S1046-2023(02)00040-3

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MeSH Terms: Apoptosis, BH3 Interacting Domain Death Agonist Protein, Carrier Proteins, Cell Death, Cell-Free System, Cytochrome c Group, Cytosol, Dose-Response Relationship, Drug, Flow Cytometry, Green Fluorescent Proteins, HeLa Cells, Humans, Immunoblotting, Jurkat Cells, Luminescent Proteins, Membrane Potentials, Microscopy, Fluorescence, Mitochondria, Potassium, Protein Transport, Proto-Oncogene Proteins, Proto-Oncogene Proteins c-bcl-2, Recombinant Fusion Proteins, Subcellular Fractions, bcl-2-Associated X Protein