Calcium and reactive oxygen species mediate staurosporine-induced mitochondrial dysfunction and apoptosis in PC12 cells

Inna Kruman, Qing Guo, Mark P. Mattson

Research output: Contribution to journalArticlepeer-review

Abstract

The bacterial alkaloid staurosporine is widely employed as an inducer of apoptosis in many cell types including neurons. The intracellular cascades that mediate staurosporine-induced apoptosis are largely unknown. Exposure of cultured PC12 cells to staurosporine resulted in a rapid (min) and prolonged (1-6 hr) elevation of intracellular free calcium levels [Ca2+](i), accumulation of mitochondrial reactive oxygen species (ROS), and decreased mitochondrial 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) reduction (1-4 hr). These early events were followed by membrane lipid peroxidation, loss of mitochondrial transmembrane potential, and nuclear apoptotic changes. Treatment of cells with serum or nerve growth factor within 1-2 hr of staurosporine exposure resulted in recovery of [Ca2+](i) and ROS levels, and rescued the cells from apoptosis. The increased [Ca2+](i) and ROS production were required for staurosporine-induced apoptosis because the intracellular calcium chelator BAPTA and uric acid (an agent that scavenges peroxynitrite) each protected cells against apoptosis. The caspase inhibitor zVAD-fmk and the anti-apoptotic gene product Bcl-2 prevented the sustained [Ca2+](i) increase and ROS accumulation induced by staurosporine indicating that caspases act very early in the apoptotic process. Our data indicate that a [Ca2+](i) increase is an early and critical event in staurosporine-induced apoptosis that engages a cell death pathway involving ROS production, oxidative stress, and mitochondrial dysfunction.

Original languageEnglish (US)
Pages (from-to)293-308
Number of pages16
JournalJournal of Neuroscience Research
Volume51
Issue number3
DOIs
StatePublished - Feb 1 1998
Externally publishedYes

Keywords

  • Caspase inhibitor
  • Fura-2
  • Lipid peroxidation
  • Nerve growth factor
  • Nitric oxide
  • Permeability transition
  • Peroxynitrite
  • Uric acid

ASJC Scopus subject areas

  • Neuroscience(all)

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