Document Detail

Mitochondrial membrane potential and nuclear changes in apoptosis caused by serum and nerve growth factor withdrawal: time course and modification by (-)-deprenyl.
MedLine Citation:
PMID:  9437015     Owner:  NLM     Status:  MEDLINE    
Studies in non-neural cells have suggested that a fall in mitochondrial membrane potential (DeltaPsiM) is one of the earliest events in apoptosis. It is not known whether neural apoptosis caused by nerve growth factor (NGF) and serum withdrawal involves a decrease in DeltaPsiM. We used epifluorescence and laser confocal microscopy with the mitochondrial potentiometric dyes chloromethyl-tetramethylrosamine methyl ester and 5,5',6, 6'-tetrachloro-1,1',3,3'-tetraethybenzimidazol carbocyanine iodide to estimate DeltaPsiM. PC12 cells were differentiated in media containing serum and NGF for 6 d before withdrawal of trophic support. After washing, the cells were incubated with media containing serum and NGF (M/S+N), media without serum and NGF, or media with the "trophic-like" monoamine oxidase B inhibitor, (-)-deprenyl. Mitochondria in cells without trophic support underwent a progressive shift to lower DeltaPsiM values that was significant by 3 hr after washing. The percentages of cells with nuclear chromatin condensation or nuclear DNA fragmentation were not significantly increased above those for cells in M/S+N until 6 hr after washing. Replacement of cells into M/S+N or treatment with (-)-deprenyl markedly reduced the proportion of mitochondria with decreased DeltaPsiM. Measurements of cytoplasmic peroxyl radical levels with 2',7'-dihydrodichlorofluorescein fluorescence and intramitochondrial Ca2+ with dihydro-rhodamine-2-acetylmethyl ester indicated that cytoplasmic peroxyl radical levels were not increased until after 6 hr, whereas increases in intramitochondrial Ca2+ paralleled the decreases in DeltaPsiM. (-)-Deprenyl appeared to alter the relationship between intramitochondrial Ca2+ levels and DeltaPsiM, possibly through its reported capacity to increase the synthesis of proteins such as BCL-2.
J S Wadia; R M Chalmers-Redman; W J Ju; G W Carlile; J L Phillips; A D Fraser; W G Tatton
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  The Journal of neuroscience : the official journal of the Society for Neuroscience     Volume:  18     ISSN:  0270-6474     ISO Abbreviation:  J. Neurosci.     Publication Date:  1998 Feb 
Date Detail:
Created Date:  1998-02-06     Completed Date:  1998-02-06     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  8102140     Medline TA:  J Neurosci     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  932-47     Citation Subset:  IM    
Department of Physiology, University of Toronto, Toronto, Ontario, Canada M55 1A8.
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MeSH Terms
Apoptosis / drug effects,  physiology*
Blood Proteins / pharmacology*
Calcium / metabolism
Cell Nucleus / drug effects,  physiology
Chromatin / physiology
Cytoplasm / metabolism
DNA Fragmentation
Free Radicals / metabolism
Image Processing, Computer-Assisted
Intracellular Membranes / physiology
Membrane Potentials / drug effects,  physiology
Microscopy, Fluorescence
Mitochondria / drug effects,  metabolism*
Nerve Growth Factors / pharmacology*
Neuroprotective Agents / pharmacology*
PC12 Cells
Peroxides / metabolism
Selegiline / pharmacology*
Reg. No./Substance:
0/Blood Proteins; 0/Chromatin; 0/Free Radicals; 0/Nerve Growth Factors; 0/Neuroprotective Agents; 0/Peroxides; 14611-51-9/Selegiline; 7440-70-2/Calcium

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