Document Detail


Synergism of nitric oxide and iron in killing the transformed murine oligodendrocyte cell line N20.1.
MedLine Citation:
PMID:  10037476     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Nitric oxide (NO) produced in inflammatory lesions may play a major role in the destruction of oligodendrocytes in multiple sclerosis and experimental allergic encephalomyelitis. The transformed murine oligodendroglial line N20.1 is much more resistant than primary oligodendrocytes to killing by the NO generator S-nitroso-N-acetyl-DL-penicillamine (SNAP). This observation prompted investigation of the mechanisms leading to cell death in the N20.1 cells and comparison of SNAP with another NO donor, sodium nitroprusside (SNP). We observed that N20.1 cells were 30 times more sensitive to SNP than to SNAP. The specific NO scavenger 2-phenyl-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide (PTIO) protected against SNP only, not against SNAP. However, dithiothreitol protected against both SNAP and SNP, indicating that S-nitrosylation of cysteines plays a major role in the cytotoxicity of both NO donors. We did not observe any formation of peroxynitrite or increase of Ca2+ concentration with either SNAP or SNP, thus excluding their involvement in the mechanisms leading to N20.1 cell death. Based on two observations, (a) potentiation of the cytotoxic effect of SNP when coincubated with ferricyanide or ferrocyanide, but not sodium cyanide, and (b) protection by deferoxamine, an iron cyanide chelator, we conclude that the greater sensitivity of N20.1 cells to SNP compared with SNAP is due to synergism between NO released and the iron cyanide portion of SNP, with the cyanide accounting for very little of the cytotoxicity. Finally, SNP but not SNAP induces some apoptosis, as shown by DNA laddering and protection by a caspase-3 inhibitor. These results suggest that low levels of NO in combination with increased iron content lead to apoptotic cell death rather than the necrotic cell death seen with higher levels of NO generated by SNAP.
Authors:
A I Boullerne; L Nedelkoska; J A Benjamins
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  Journal of neurochemistry     Volume:  72     ISSN:  0022-3042     ISO Abbreviation:  J. Neurochem.     Publication Date:  1999 Mar 
Date Detail:
Created Date:  1999-03-18     Completed Date:  1999-03-18     Revised Date:  2007-11-14    
Medline Journal Info:
Nlm Unique ID:  2985190R     Medline TA:  J Neurochem     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  1050-60     Citation Subset:  IM    
Affiliation:
Department of Neurology, Wayne State University, School of Medicine, Detroit, Michigan, USA.
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MeSH Terms
Descriptor/Qualifier:
Animals
Cell Death / drug effects
Cell Line
Coloring Agents
Culture Media, Serum-Free
DNA Fragmentation / drug effects
Drug Synergism
Immunohistochemistry
Iron / antagonists & inhibitors,  toxicity*
Kinetics
Mice
Nitrates / metabolism
Nitric Oxide / antagonists & inhibitors,  physiology*,  toxicity
Nitric Oxide Donors / antagonists & inhibitors,  toxicity
Nitroprusside / toxicity
Oligodendroglia / drug effects*
Oxidants / metabolism
Penicillamine / analogs & derivatives,  antagonists & inhibitors,  toxicity
S-Nitroso-N-Acetylpenicillamine
Trypan Blue
Grant Support
ID/Acronym/Agency:
NS13143/NS/NINDS NIH HHS
Chemical
Reg. No./Substance:
0/Coloring Agents; 0/Culture Media, Serum-Free; 0/Nitrates; 0/Nitric Oxide Donors; 0/Oxidants; 10102-43-9/Nitric Oxide; 15078-28-1/Nitroprusside; 26404-66-0/peroxynitric acid; 52-67-5/Penicillamine; 72-57-1/Trypan Blue; 7439-89-6/Iron; 79032-48-7/S-Nitroso-N-Acetylpenicillamine

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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