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Calcium and reactive oxygen species mediate staurosporine‐induced mitochondrial dysfunction and apoptosis in PC12 cells
Author(s) -
Kruman Inna,
Guo Qing,
Mattson Mark P.
Publication year - 1998
Publication title -
journal of neuroscience research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.72
H-Index - 160
eISSN - 1097-4547
pISSN - 0360-4012
DOI - 10.1002/(sici)1097-4547(19980201)51:3<293::aid-jnr3>3.0.co;2-b
Subject(s) - staurosporine , apoptosis , reactive oxygen species , microbiology and biotechnology , bapta , calcium in biology , programmed cell death , mitochondrion , oxidative stress , intracellular , biology , mitochondrial ros , chemistry , biochemistry , protein kinase c , signal transduction
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 [Ca 2+ ] 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 [Ca 2+ ] i and ROS levels, and rescued the cells from apoptosis. The increased [Ca 2+ ] 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 [Ca 2+ ] i increase and ROS accumulation induced by staurosporine indicating that caspases act very early in the apoptotic process. Our data indicate that a [Ca 2+ ] 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. J. Neurosci. Res. 51:293–308, 1998. © 1998 Wiley‐Liss, Inc.