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

J. H M Prehn*, J. Jordán, G. D. Ghadge, E. Preis, M. F. Galindo, R. P. Roos, J. Krieglstein, R. J. Miller

*Corresponding author for this work

Research output: Contribution to journalArticle

115 Scopus citations

Abstract

Staurosporine (0.03-0.5 μM) 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 μM) or the cell cycle inhibitor mimosine (100 μM). To investigate the role of Ca2+ and reactive oxygen species in staurosporine-induced neuronal apoptosis, we overexpressed calbindin D(28K), 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 D(28K) and Cu/Zn superoxide dismutase significantly reduced staurosporine neurotoxicity compared with control cultures infected with a β-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 μM) nor the NMDA receptor antagonist dizocilpine (MK-801; 1 μM) was able to reduce staurosporine neurotoxicity. Cultures treated with the antioxidants U- 74500A (1-10 μM) and N-acetylcysteine (100 μM) also demonstrated reduced staurosporine neurotoxicity. These results suggest a fundamental role for both Ca2+ and reactive oxygen species in staurosporine-induced neuronal apoptosis.

Original languageEnglish (US)
Pages (from-to)1679-1685
Number of pages7
JournalJournal of neurochemistry
Volume68
Issue number4
StatePublished - Apr 1 1997

Keywords

  • Adenovirus
  • Antioxidants
  • Calbindin D(28K)
  • NMDA
  • Programmed cell death
  • Superoxide dismutase

ASJC Scopus subject areas

  • Biochemistry
  • Cellular and Molecular Neuroscience

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    Prehn, J. H. M., Jordán, J., Ghadge, G. D., Preis, E., Galindo, M. F., Roos, R. P., Krieglstein, J., & Miller, R. J. (1997). Ca2+ and reactive oxygen species in staurosporine-induced neuronal apoptosis. Journal of neurochemistry, 68(4), 1679-1685.