Production of reactive oxygen species and release of L-glutamate during superoxide anion-induced cell death of cerebellar granule neurons.
Enhanced production of superoxide anion (O2-) is considered to play a pivotal role in the pathogenesis of CNS neurons. Here, we report that O2- generated by xanthine (XA) + xanthine oxidase (XO) triggered cell death associated with nuclear condensation and DNA fragmentation in cerebellar granule neuron. XA + XO induced significant increases in amounts of intracellular reactive oxygen species (ROS) before initiating loss of cell viability, as determined by measurement of 6-carboxy-2',7'-dichlorodihydrofluorescein diacetate, di(acetoxymethyl ester) (C-DCDHF-DA) for O2- and other ROS and hydroethidine (HEt) specifically for O2- by using fluorescence microscopy and flow cytometry. Catalase, but not superoxide dismutase (SOD), significantly protected granule neurons from the XA + XO-induced cell death. Catalase effectively reduced C-DCDHF-DA but not HEt fluorescence, whereas SOD reduced HEt but not C-DCDHF-DA fluorescence, indicating that HEt and C-DCDHF-DA fluorescence correlated with O2- and hydrogen peroxide, respectively. The NMDA antagonist MK-801 prevented the death. XA + XO induced an increase in L-glutamate release from cerebellar granule neurons. These results indicate that elevation of O2- induces cell death associated with increasing ROS production in cerebellar granule neurons and that XA + XO enhanced release of L-glutamate.
Satoh
T
T
Division of Protein Biosynthesis, Institute for Protein Research, Osaka University, Japan.
Numakawa
T
T
Abiru
Y
Y
Yamagata
T
T
Ishikawa
Y
Y
Enokido
Y
Y
Hatanaka
H
H
D000818:Animals; D000838:Anions / Q000494:pharmacology*; D016923:Cell Death / Q000502:physiology; D002478:Cells, Cultured; D002531:Cerebellum / Q000166:cytology / Q000378:metabolism*; D004338:Drug Combinations; D018698:Glutamic Acid / Q000378:metabolism*; D009474:Neurons / Q000502:physiology*; D051381:Rats; D017382:Reactive Oxygen Species / Q000378:metabolism*; D013481:Superoxides / Q000494:pharmacology*; D019820:Xanthine / Q000494:pharmacology; D014969:Xanthine Oxidase / Q000494:pharmacology
D000838:Anions; D004338:Drug Combinations; D017382:Reactive Oxygen Species; D013481:Superoxides; D019820:Xanthine; D018698:Glutamic Acid; D014969:Xanthine Oxidase