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The Journal of Neuroscience, January 1, 2000, 20(1):259-265
Proteasome Inhibitors Induce Cytochrome c-Caspase-3-Like Protease-Mediated Apoptosis in Cultured Cortical Neurons
Jian Hua Qiu1, Akio Asai1, 3, Shunji Chi1, 3, Nobuhito Saito1, Hirofumi Hamada2, and Takaaki Kirino1, 3 1 Laboratory for Neuroscience and Neurooncology, Faculty of Medicine, University of Tokyo, Tokyo, 113-8655 Japan, 2 Department of Molecular Biotherapy Research, Cancer Chemotherapy Center, Cancer Institute, Tokyo, 170-8455 Japan, and 3 CREST (Core Research for Evolutional Science and Technology), Japan Science and Technology Corporation, Kawaguchi, 332-0012 Japan
The ubiquitin-proteasome protein degradation pathway is crucial in controlling intracellular levels of a variety of short-lived proteins and maintaining cellular growth and metabolism. In a previous study, we showed the accumulation of conjugated ubiquitin in CA1 neurons of the gerbil after 5 min of forebrain ischemia (Morimoto et al., 1996; Ide et al., 1999). The accumulation of conjugated ubiquitin may reflect proteasome malfunction. In the present study, we investigated the effects of proteasome inhibitors on primary neuronal cultures to determine whether proteasomal malfunction induces neuronal death. When carbobenzoxy-Leu-Leu-Leu-aldehyde or lactacystin, two different types of proteasome inhibitors, were separately used to suppress proteasome activity, we observed induction of apoptotic neuronal cell death in both cases. During the apoptotic process, mitochondrial membrane potential was disrupted, cytochrome-c was released from mitochondria into the cytosol, and caspase-3-like proteases were activated. Apoptosis was inhibited by pretreatment with acetyl-aspartyl-glutamyl-valyl-aspart-1-aldehyde or overexpression of Bcl-x/L. These results demonstrated that suppression of proteasome function induces neuronal apoptosis via the release of cytochrome c from mitochondria and activation of caspase-3-like proteases.
Key words: neuron; apoptosis; proteasome; cytochrome c; caspase-3-like proteases; ubiquitin
Copyright © 2000 Society for Neuroscience 0270-6474/0/201259-07$05.00/0
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