Jurkat T cells showed a major, early decrease in blue autofluorescence in response to Fas/Apo-1/CD95 cross-linking or stimulation with cell-permeant ceramide. This indicates the oxidation/depletion of NADH or NADPH before the onset of apoptosis. Kinetic studies, cytofluorimetric multiparameter analyses and cell sorting experiments indicated a close temporal relationship between NAD(P)H oxidation/depletion and the dissipation of the mitochondrial transmembrane potential (∆Ψm). In contrast, NAD(P)H depletion was detected well before several other changes associated with late apoptosis, including enhanced superoxide generation, phosphatidylserine exposure on the cell surface, loss of cytosolic K+, decreased cytoplasmic pH, nuclear DNA fragmentation, cell shrinkage, loss of viability and the appearance of the mitochondrial antigen APO2.7. Full activation of caspase 9 and caspase 3 appeared to be correlated with the appearance of superoxide anions in the mitochondria, and followed the drop in NADPH. Overexpression of the apoptosis-inhibitory proto-oncogene Bcl-2, which encodes an inhibitor of the mitochondrial permeability transition (PT) pore, delayed both the ∆Ψm disruption and the depletion of NAD(P)H. Similar effects were observed with the pharmacological PT pore inhibitors, bongkrekic acid and cyclosporin A. Thus there appears to be a close functional relationship between mitochondrial and cellular redox changes during early apoptosis; events that are inhibited by Bcl-2.
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January 2001
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Research Article|
January 08 2001
Oxidation of pyridine nucleotides during Fas- and ceramide-induced apoptosis in Jurkat cells: correlation with changes in mitochondria, glutathione depletion, intracellular acidification and caspase 3 activation
Patrice Xavier PETIT;
Patrice Xavier PETIT
1
*Institut Cochin de Génétique Moléculaire, INSERM U129, CHU Cochin Port-Royal, 24 rue du Faubourg Saint-Jacques, F-75014 Paris, France,
‡INSERM U131, IPSC, 32 rue des Carnets, 92140 Clamart, France,
1To whom correspondence should be addressed (e-mail pxpetit@icgm.cochin.inserm.fr).
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Marie-Claude GENDRON;
Marie-Claude GENDRON
†Service de Cytométrie en Flux, Institut Jacques Monod, Université Denis Diderot, 4 place Jussieu, F-75005 Paris, France,
‡INSERM U131, IPSC, 32 rue des Carnets, 92140 Clamart, France,
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Nicolas SCHRANTZ;
Nicolas SCHRANTZ
*Institut Cochin de Génétique Moléculaire, INSERM U129, CHU Cochin Port-Royal, 24 rue du Faubourg Saint-Jacques, F-75014 Paris, France,
‡INSERM U131, IPSC, 32 rue des Carnets, 92140 Clamart, France,
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Didier MÉTIVIER;
Didier MÉTIVIER
§Centre National de la Recherche Scientifique, UPR 420, 19 rue Guy Môquet, F-94801 Villejuif, France
‡INSERM U131, IPSC, 32 rue des Carnets, 92140 Clamart, France,
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Guido KROEMER;
Guido KROEMER
§Centre National de la Recherche Scientifique, UPR 420, 19 rue Guy Môquet, F-94801 Villejuif, France
‡INSERM U131, IPSC, 32 rue des Carnets, 92140 Clamart, France,
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Zofia MACIOROWSKA;
Zofia MACIOROWSKA
ǁInstitut Curie, Département de Pathologie, 26 rue d'Ulm, 75248 Paris Cedex 05, France
‡INSERM U131, IPSC, 32 rue des Carnets, 92140 Clamart, France,
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Franck SUREAU;
Franck SUREAU
*Institut Cochin de Génétique Moléculaire, INSERM U129, CHU Cochin Port-Royal, 24 rue du Faubourg Saint-Jacques, F-75014 Paris, France,
‡INSERM U131, IPSC, 32 rue des Carnets, 92140 Clamart, France,
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Steven KOESTER
Steven KOESTER
¶Beckman Coulter, Inc., Advanced Technology, 11800 S.W. 147th Avenue, Miami, FL 33196-2500, U.S.A.
‡INSERM U131, IPSC, 32 rue des Carnets, 92140 Clamart, France,
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Publisher: Portland Press Ltd
Received:
April 26 2000
Revision Received:
September 28 2000
Accepted:
October 25 2000
Online ISSN: 1470-8728
Print ISSN: 0264-6021
The Biochemical Society, London © 2001
2001
Biochem J (2001) 353 (2): 357–367.
Article history
Received:
April 26 2000
Revision Received:
September 28 2000
Accepted:
October 25 2000
Citation
Patrice Xavier PETIT, Marie-Claude GENDRON, Nicolas SCHRANTZ, Didier MÉTIVIER, Guido KROEMER, Zofia MACIOROWSKA, Franck SUREAU, Steven KOESTER; Oxidation of pyridine nucleotides during Fas- and ceramide-induced apoptosis in Jurkat cells: correlation with changes in mitochondria, glutathione depletion, intracellular acidification and caspase 3 activation. Biochem J 15 January 2001; 353 (2): 357–367. doi: https://doi.org/10.1042/bj3530357
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