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Detection of Superoxide Production by Activated Microglia Using a Sensitive and Specific Chemiluminescence Assay and Microglia‐Mediated PC12h Cell Death
Author(s) -
Tanaka M.,
Sotomatsu A.,
Yoshida T.,
Hirai S.,
Nishida A.
Publication year - 1994
Publication title -
journal of neurochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.75
H-Index - 229
eISSN - 1471-4159
pISSN - 0022-3042
DOI - 10.1046/j.1471-4159.1994.63010266.x
Subject(s) - microglia , superoxide , superoxide dismutase , chemiluminescence , reactive oxygen species , chemistry , programmed cell death , cytotoxicity , neuroglia , phorbol , microbiology and biotechnology , biochemistry , biology , immunology , oxidative stress , endocrinology , in vitro , central nervous system , inflammation , apoptosis , signal transduction , enzyme , organic chemistry , protein kinase c
Superoxide production by cultured microglia derived from neonatal rat brains and the cytotoxicity of these cells were evaluated. The chemiluminescence (photon counts) detected in the presence of MCLA, a new chemiluminescence probe, was strongly correlated with the microglial cell count. Chemiluminescence observed in this system was confirmed to originate specifically from superoxide produced by activated microglia. Phorbol myristate acetate‐stimulated microglia caused a pronounced reduction of PC12h cell numbers in coculture. The addition of superoxide dismutase with catalase or the addition of deferoxamine mesylate inhibited PC12h cell death, suggesting that active oxygen species derived from superoxide generated by the microglia or iron‐oxygen complex formation were responsible for the cytotoxicity. These results imply that activated microglia may participate in the progression of the pathologic process in some neurodegenerative disorders.