title: Confocal microscopy of the mitochondrial permeability transition in necrotic and apoptotic cell death.
abstract: Opening of a high-conductance pore in the mitochondrial inner membrane induces onset of the mitochondrial permeability transition (mPT). Cyclosporin A and trifluoperazine inhibit this pore and block necrotic cell death in oxidative stress, Ca2+ ionophore toxicity, Reye-related drug toxicity, pH-dependent ischaemia/reperfusion injury and other models of cell injury. Confocal fluorescence microscopy directly visualizes the increased mitochondrial membrane permeability of the mPT from the movement of calcein from the cytosol into the matrix space. Pyridine nucleotide oxidation, increased mitochondrial Ca2+ and mitochondrial generation of reactive oxygen species (ROS) all contribute to the onset of the mPT in situ. Confocal microscopy also shows directly that the mPT is a critical link in apoptotic signalling by tumour necrosis factor-alpha at a point downstream of caspase 8 and upstream of caspase 3. Cyclosporin A blocks this mPT, preventing release of pro-apoptotic cytochrome c from mitochondria and subsequent apoptotic cell killing. Progression to necrosis or apoptosis after the mPT depends on the availability of ATP, which blocks necrosis but promotes the apoptotic programme. Given the pathophysiological importance of the mPT, development of agents to modulate the mPT represents an important new goal for pharmaceutical drug discovery.
authors.affiliation: Department of Cell Biology and Anatomy, University of North Carolina at Chapel Hill 27599-7090, USA.
authors.forename: J J
authors.initials: JJ
authors.lastname: Lemasters
authors.forename: T
authors.initials: T
authors.lastname: Qian
authors.forename: L C
authors.initials: LC
authors.lastname: Trost
authors.forename: B
authors.initials: B
authors.lastname: Herman
authors.forename: W E
authors.initials: WE
authors.lastname: Cascio
authors.forename: C A
authors.initials: CA
authors.lastname: Bradham
authors.forename: D A
authors.initials: DA
authors.lastname: Brenner
authors.forename: A L
authors.initials: AL
authors.lastname: Nieminen
mesh_terms: D000818:Animals; D017209:Apoptosis; D002463:Cell Membrane Permeability; D018613:Microscopy, Confocal; D008928:Mitochondria / Q000502:physiology*; D009336:Necrosis