Distinct alterations in mitochondrial mass and function characterize different models of apoptosis.
Recent studies have shown that reduction in mitochondrial membrane potential (delta psi m) and generation of reactive oxygen species are early events in apoptosis. In this study, we present two different models of apoptotic cell death, Chinese hamster ovary (CHO) cells treated with aphidicolin and dexamethasone-treated 2B4 T-cell hybridoma cells, which display opposing mitochondrial changes. CHO cells arrested at G1/S with aphidicolin have a progressive increase in mitochondria mass and number, assessed by flow cytometry and fluorescent microscopy with mitochondria-specific probes. The increase in mitochondrial mass was not accompanied by a gain in net cellular mitochondrial membrane potential, consistent with an accumulation of relatively depolarized mitochondria. Fluorescent microscopy demonstrated an increased content of low delta psi m mitochondria in aphidicolin-treated CHO cells, but high delta psi m mitochondria were also present and remained stable in number. Mitochondrial mass correlated with decreased clonogenicity of aphidicolin-treated CHO cells. Cycloheximide prevented both the proliferation of mitochondria and subsequent cell death. In contrast, dexamethasone treatment of 2B4 T-cell hybridoma cells caused a decrease in delta psi m without mitochondrial proliferation. Cycloheximide and Bcl-2 overexpression inhibited the loss of delta psi m, as well as apoptosis. In both models, cell death was associated with a decrease in mitochondrial potential relative to mitochondrial mass, suggesting that an accumulation of damaged or dysfunctional mitochondria had occurred.
Camilleri-Broët
S
S
Division of Clinical Research and Molecular Medicine, Fred Hutchinson Cancer Research Center, Seattle, Washington 98109, USA.
Vanderwerff
H
H
Caldwell
E
E
Hockenbery
D
D
D000818:Animals; D016590:Aphidicolin / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects / Q000502:physiology*; D016466:CHO Cells / Q000187:drug effects; D002470:Cell Survival; D003114:Colony-Forming Units Assay; D006224:Cricetinae; D003513:Cycloheximide / Q000494:pharmacology; D003907:Dexamethasone / Q000494:pharmacology; D005434:Flow Cytometry; D019254:Genes, bcl-2; D006825:Hybridomas / Q000187:drug effects; D008854:Microscopy, Electron; D008856:Microscopy, Fluorescence; D008928:Mitochondria / Q000378:metabolism*; D008954:Models, Biological; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology; D013601:T-Lymphocytes / Q000187:drug effects; D014162:Transfection
D011500:Protein Synthesis Inhibitors; D016590:Aphidicolin; D003907:Dexamethasone; D003513:Cycloheximide