Quantitation of mitochondrial alterations associated with apoptosis

Maria Castedo, Karine Ferri, Thomas Roumier, Didier Métivier, Naoufal Zamzami, Guido Kroemer

Résultats de recherche: Contribution à un journalArticleRevue par des pairs

251 Citations (Scopus)

Résumé

Mitochondria undergo two major changes during early apoptosis. On the one hand, the outer mitochondrial membrane becomes permeable to proteins, resulting in the release of soluble intermembrane proteins (SIMPs) from the mitochondrion. On the other hand, the inner mitochondrial membrane transmembrane potential (ΔΨm) is reduced. These changes occur in most, if not all, models of cell death and can be taken advantage of to detect apoptosis at an early stage. Here, we delineate methods for the detection of alterations in the ΔΨm, based on the incubation of cells with cationic lipophilic fluorochromes, the uptake of which is driven by the ΔΨm. Certain ΔΨm-sensitive dyes can be combined with other fluorochromes to detect simultaneously cellular viability, plasma membrane exposure of phosphatidylserine residues, or the mitochondrial production of reactive oxygen species (ROS). In addition, we describe an immunofluorescence method for the detection of two functionally important proteins translocating from mitochondria, namely, the caspase co-activator cytochrome c and the caspase-independent death effector apoptosis inducing factor (AIF).

langue originaleAnglais
Pages (de - à)39-47
Nombre de pages9
journalJournal of Immunological Methods
Volume265
Numéro de publication1-2
Les DOIs
étatPublié - 1 juil. 2002
Modification externeOui

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