Involvement of cystatin C in oxidative stress-induced apoptosis of cultured rat CNS neurons.
Oxidative stress is involved in neuronal degeneration in cerebrovascular injury, neuropathology and aging. When rat CNS neurons were cultured in a high (50%) oxygen atmosphere, the neurons died. This high oxygen-induced cell death showed features of apoptotic cell death, characterized by DNA fragmentation, and was blocked by inhibitor of protein synthesis. We found that cystatin C and HuC mRNA, the products of which are an inhibitor of cysteine proteases and an RNA binding protein, respectively, were up-regulated in neurons cultured in the high oxygen atmosphere. In the present study, we focused on cystatin C. Cystatin C protein levels were also increased in neurons cultured in the high oxygen atmosphere. In situ hybridization with an RNA probe for rat cystatin C and immunocytochemistry with anti-human cystatin C antibody showed that microtubule-associated protein 2 (MAP2)-positive neurons expressed cystatin C mRNA and protein, respectively, in the high oxygen atmosphere. These results indicated that oxidative stress stimulates an increase in cystatin C expression in cultured neurons, and that cystatin C might have important roles in regulation of apoptosis elicited by oxidative stress.
Institute for Protein Research, Osaka University, 3-2 Yamadaoka, Suita, 565-0871, Osaka, Japan.
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Nishio
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Yoshida
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Nishiyama
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Hatanaka
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Yamada
D000818:Animals; D017209:Apoptosis / Q000502:physiology*; D002478:Cells, Cultured; D002490:Central Nervous System / Q000201:enzymology* / Q000473:pathology / Q000503:physiopathology; D055316:Cystatin C; D015891:Cystatins / Q000235:genetics* / Q000378:metabolism*; D003546:Cysteine Endopeptidases / Q000378:metabolism; D005260:Female; D005333:Fetus; D005796:Genes / Q000502:physiology; D008297:Male; D008869:Microtubule-Associated Proteins / Q000378:metabolism; D009474:Neurons / Q000201:enzymology* / Q000473:pathology; D018384:Oxidative Stress / Q000502:physiology*; D010100:Oxygen / Q000009:adverse effects*; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D015854:Up-Regulation / Q000502:physiology
C525030:CST3 protein, human; C525032:Cst3 protein, rat; D055316:Cystatin C; D015891:Cystatins; D008869:Microtubule-Associated Proteins; D012333:RNA, Messenger; D003546:Cysteine Endopeptidases; D010100:Oxygen