Abstract
The relationship is investigated between mitochondrial membrane potential (ΔΨM), respiration and cytochrome c (cyt c) release in single neural bcl-2 transfected cells (GT1-7bcl-2) or GT1-7puro cells during apoptosis induced by staurosporine (STS). Bcl-2 inhibited the mitochondrial release of cyt c and apoptosis. Three different cell responses to STS were identified in GT1-7puro cells: (i) neither ΔΨM nor cyt c were significantly affected; (ii) a decrease in ΔΨM was accompanied by a complete release of cyt c; or (iii) cyt c release occurred independently of a loss of ΔΨM. The endogenous inner membrane proton leak of the in situ mitochondria, monitored by respiration in the presence of oligomycin, was increased by STS by 92% in puro cells, but by only 23% in bcl-2 cells. STS decreased respiratory capacity, in the presence of protonophore, by 31% in puro cells and by 20% in bcl-2 cells. In the absence of STS, oligomycin hyperpolarized mitochondria within both puro and bcl-2-transfected cells, indicating that the organelles were net generators of ATP. However after 15 h exposure to STS oligomycin rapidly collapsed residual mitochondrial polarization in the puro cells, indicating that ΔΨM had been maintained by ATP synthase reversal. bcl-2 cells in contrast, maintained ΔΨM until protonophore was added. These results indicate that the maintenance of ΔΨM following release of cyt c may be a consequence of ATP synthase reversal and cytoplasmic ATP hydrolysis in STS-treated GT1-7 cells. Cell Death and Differentiation (2001) 8, 995–1003
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Abbreviations
- FCCP:
-
carbonylcyanide-p-(trifluoromethoxy) phenylhydrazone
- STS:
-
staurosporine
- ΔΨM:
-
mitochondrial membrane potential
- TMRM+:
-
tetramethylrhodamine methyl ester
- PS:
-
phosphatidylserine
- cyt c:
-
cytochrome c
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Rego, A., Vesce, S. & Nicholls, D. The mechanism of mitochondrial membrane potential retention following release of cytochrome c in apoptotic GT1-7 neural cells. Cell Death Differ 8, 995–1003 (2001). https://doi.org/10.1038/sj.cdd.4400916
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DOI: https://doi.org/10.1038/sj.cdd.4400916
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