Despite the internucleosomal cleavage of DNA, reactive oxygen species do not produce other markers of apoptosis in cultured neurons.
The cell death induced by hydroxyl radicals generated by Cu-phenanthroline and peroxynitrite generated by 3-morpholinosydnonimine hydrochloride (SIN-1) in rat primary cortical neuronal cultures was compared with the apoptotic death induced by staurosporine and the necrotic death induced by glutamate. Both SIN-1 and Cu-phenanthroline were capable of generating internucleosomal cleavage of DNA-a hallmark of apoptosis. Other characteristics of this cell death, such as nuclear morphology by light microscopy; DNA breaks by single-cell gel electrophoresis; the effects of the apoptotic inhibitors cycloheximide, aurintricarboxylic acid, and tosyl-l-lysine chloromethyl ketone; the measurement of caspase activity; and the effects of antioxidants, were then analyzed. The conclusion from these hallmarks of apoptosis is that the cell death induced by these reactive oxygen species is not apoptosis.
Hill
I E
IE
Institute for Biological Sciences, National Research Council of Canada, Ottawa, Ontario, K1A 0R6, Canada.
Murray
C
C
Richard
J
J
Rasquinha
I
I
MacManus
J P
JP
D000818:Animals; D000975:Antioxidants / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects / Q000502:physiology*; D015415:Biomarkers; D020169:Caspases / Q000378:metabolism; D002467:Cell Nucleus / Q000473:pathology; D002800:Cholinesterase Inhibitors / Q000494:pharmacology; D020552:Comet Assay; D003513:Cycloheximide / Q000494:pharmacology; D053938:DNA Fragmentation / Q000187:drug effects*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005260:Female; D016166:Free Radical Scavengers / Q000494:pharmacology; D018698:Glutamic Acid / Q000494:pharmacology; D008665:Metalloporphyrins / Q000494:pharmacology; D008981:Molsidomine / Q000031:analogs & derivatives / Q000494:pharmacology; D009410:Nerve Degeneration / Q000139:chemically induced / Q000378:metabolism; D009419:Nerve Tissue Proteins / Q000378:metabolism; D009474:Neurons / Q000166:cytology* / Q000201:enzymology; D009707:Nucleosomes / Q000378:metabolism*; D010084:Oxidation-Reduction; D010618:Phenanthrolines / Q000494:pharmacology; D011247:Pregnancy; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology; D051381:Rats; D017207:Rats, Sprague-Dawley; D017382:Reactive Oxygen Species / Q000378:metabolism*; D019311:Staurosporine / Q000494:pharmacology; D014527:Uric Acid / Q000494:pharmacology
D000975:Antioxidants; D015415:Biomarkers; D002800:Cholinesterase Inhibitors; D004791:Enzyme Inhibitors; D016166:Free Radical Scavengers; D008665:Metalloporphyrins; D009419:Nerve Tissue Proteins; D009707:Nucleosomes; D010618:Phenanthrolines; D011500:Protein Synthesis Inhibitors; D017382:Reactive Oxygen Species; C097284:manganese(III)-tetrakis(4-benzoic acid)porphyrin; D014527:Uric Acid; D018698:Glutamic Acid; C002385:linsidomine; C050950:tetrakis(N-methyl-4-pyridiniumyl)porphine manganese(III) complex; D003513:Cycloheximide; D008981:Molsidomine; D020169:Caspases; D019311:Staurosporine; C025205:1,10-phenanthroline