Ca2+ and reactive oxygen species in staurosporine-induced neuronal apoptosis.
Staurosporine (0.03-0.5 microM) induced a dose-dependent, apoptotic degeneration in cultured rat hippocampal neurons that was sensitive to 24-h pretreatments with the protein synthesis inhibitor cycloheximide (1 microM) or the cell cycle inhibitor mimosine (100 microM). To investigate the role of Ca2+ and reactive oxygen species in staurosporine-induced neuronal apoptosis, we overexpressed calbindin D28K, a Ca2+ binding protein, and Cu/ Zn superoxide dismutase, an antioxidative enzyme, in the hippocampal neurons using adenovirus-mediated gene transfer. Infection of the cultures with the recombinant adenoviruses (100 multiplicity of infection) resulted in a stable expression of the respective proteins assessed 48 h later. Overexpression of both calbindin D28K and Cu/Zn superoxide dismutase significantly reduced staurosporine neurotoxicity compared with control cultures infected with a beta-galactosidase overexpressing adenovirus. Staurosporine-induced neuronal apoptosis was also significantly reduced when the culture medium was supplemented with 10 or 30 mM K+, suggesting that Ca2+ influx via voltage-sensitive Ca2+ channels reduces this apoptotic cell death. In contrast, neither the glutamate receptor agonist NMDA (1-10 microM) nor the NMDA receptor antagonist dizocilpine (MK-801; 1 microM) was able to reduce staurosporine neurotoxicity. Cultures treated with the antioxidants U-74500A (1-10 microM) and N-acetylcysteine (100 microM) also demonstrated reduced staurosporine neurotoxicity. These results suggest a fundamental role for both Ca2+ and reactive oxygen species in staurosprine-induced neuronal apoptosis.
Prehn
J H
JH
Department of Pharmacology and Toxicology, Philipps-University, Marburg, Germany.
Jordán
J
J
Ghadge
G D
GD
Preis
E
E
Galindo
M F
MF
Roos
R P
RP
Krieglstein
J
J
Miller
R J
RJ
D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D064092:Calbindin 1; D064026:Calbindins; D002118:Calcium / Q000032:analysis*; D015220:Calcium Channels / Q000502:physiology; D002478:Cells, Cultured / Q000166:cytology / Q000187:drug effects / Q000201:enzymology; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D005609:Free Radicals / Q000378:metabolism; D015870:Gene Expression / Q000187:drug effects; D006624:Hippocampus / Q000166:cytology; D015640:Ion Channel Gating / Q000502:physiology; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000378:metabolism; D009474:Neurons / Q000737:chemistry / Q000166:cytology* / Q000201:enzymology; D009498:Neurotoxins / Q000494:pharmacology; D051381:Rats; D017207:Rats, Sprague-Dawley; D017382:Reactive Oxygen Species / Q000378:metabolism*; D016194:Receptors, N-Methyl-D-Aspartate / Q000502:physiology; D064030:S100 Calcium Binding Protein G / Q000235:genetics / Q000378:metabolism; D019311:Staurosporine / Q000494:pharmacology*; D013482:Superoxide Dismutase / Q000378:metabolism
C576500:Calb1 protein, rat; D064092:Calbindin 1; D064026:Calbindins; D015220:Calcium Channels; D004791:Enzyme Inhibitors; D005609:Free Radicals; D009419:Nerve Tissue Proteins; D009498:Neurotoxins; D017382:Reactive Oxygen Species; D016194:Receptors, N-Methyl-D-Aspartate; D064030:S100 Calcium Binding Protein G; D013482:Superoxide Dismutase; D019311:Staurosporine; D002118:Calcium