Abstract
Isolated mitochondria are an invaluable analytical tool to probe mitochondrial function and evaluate apoptosis induction via the so-called mitochondrial pathway. Irrespective of their tissue origin (e.g., heart, liver, muscle, brain), these organelles participate actively to cell and life decision by producing energy for cell metabolism, but also by undergoing a lethal and irreversible mitochondrial membrane permeabilization (MMP) in stress and pathological conditions. MMP consequences consist, at least in part, in loss of mitochondrial transmembrane potential (ΔΨm), matrix swelling, arrest of respiration and ATP production, and cytochrome c release from the intermembrane space to the cytosol. These parameters can be evaluated in vitro via several miniaturized assays, which have tremendous applications in the field of pharmacology, toxicology, diagnosis, as well as drug discovery.
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References
Martinou JC (1999) Apoptosis—key to the mitochondrial gate. Nature 399:411–412
Brenner C, Kroemer G (2000) Apoptosis. Mitochondria—the death signal integrators. Science 289:1150–1151
Kroemer G, Galluzzi L, Brenner C (2007) Mitochondrial membrane permeabilization in cell death. Physiol Rev 87:99–163
Zamzami N, Marchetti P, Castedo M, Decaudin D, Macho A, Hirsch T, Susin SA et al (1995) Sequential reduction of mitochondrial transmembrane potential and generation of reactive oxygen species in early programmed cell death. J Exp Med 182:367–377
Gogvadze V, Robertson JD, Zhivotovsky B, Orrenius S (1996) Cytochrome c release occurs via Ca2+-dependent and Ca2+-independent mechanisms that are regulated by Bax. J Biol Chem 276:19066–19071
Liu X, Kim CN, Yang J, Jemmerson R, Wang X (1996) Induction of apoptic program in cell-free extracts: requirement for dATP and cytochrome C. Cell 86:147–157
Crompton M, McGuinness O, Nazareth W (1992) The involvement of cyclosporin A binding proteins in regulating and uncoupling mitochondrial energy transduction. Biochim Biophys Acta 1101:214–217
Belzacq-Casagrande A, Martel C, Pertuiset C, Borgne-Sanchez A, Jacotot E, Brenner C (2009) Pharmacological screening and enzymatic assays for apoptosis. Front Biosci 14:3550–3562
Porceddu M, Buron N, Roussel C, Labbe G, Fromenty B, Borgne-Sanchez A (2012) Prediction of liver injury induced by chemicals in human with a multiparametric assay on isolated mouse liver mitochondria. Toxicol Sci 129:332–345
Martel C, Allouche M, Esposti DD, Fanelli E, Boursier C, Henry C, Chopineau J et al (2013) GSK3-mediated VDAC phosphorylation controls outer mitochondrial membrane permeability during lipid accumulation. Hepatology 57:93–102
Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685
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Wang, Z., Nicolas, C., Fischmeister, R., Brenner, C. (2015). Enzymatic Assays for Probing Mitochondrial Apoptosis. In: Weissig, V., Edeas, M. (eds) Mitochondrial Medicine. Methods in Molecular Biology, vol 1265. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2288-8_30
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DOI: https://doi.org/10.1007/978-1-4939-2288-8_30
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-2287-1
Online ISBN: 978-1-4939-2288-8
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