9084441

pubmed:Citation

4

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.

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MEDLINE

0022-3042

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NLM

1679-85

pubmed-meshheading:9084441-Animals, pubmed-meshheading:9084441-Apoptosis, pubmed-meshheading:9084441-Calcium, pubmed-meshheading:9084441-Calcium Channels, pubmed-meshheading:9084441-Calcium-Binding Protein, Vitamin D-Dependent, pubmed-meshheading:9084441-Cells, Cultured, pubmed-meshheading:9084441-Enzyme Inhibitors, pubmed-meshheading:9084441-Free Radicals, pubmed-meshheading:9084441-Gene Expression, pubmed-meshheading:9084441-Hippocampus, pubmed-meshheading:9084441-Ion Channel Gating, pubmed-meshheading:9084441-Nerve Tissue Proteins, pubmed-meshheading:9084441-Neurons, pubmed-meshheading:9084441-Neurotoxins, pubmed-meshheading:9084441-Rats, pubmed-meshheading:9084441-Rats, Sprague-Dawley, pubmed-meshheading:9084441-Reactive Oxygen Species, pubmed-meshheading:9084441-Receptors, N-Methyl-D-Aspartate, pubmed-meshheading:9084441-Staurosporine, pubmed-meshheading:9084441-Superoxide Dismutase

Ca2+ and reactive oxygen species in staurosporine-induced neuronal apoptosis.

Journal Article, Research Support, U.S. Gov't, P.H.S., Research Support, Non-U.S. Gov't