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Originally published In Press as doi:10.1074/jbc.M204176200 on August 22, 2002
J. Biol. Chem., Vol. 277, Issue 44, 41455-41462, November 1, 2002
A Newly Established Neuronal  -0 Cell Line Highly Susceptible
to Oxidative Stress Accumulates Iron and Other Metals
RELEVANCE TO THE ORIGIN OF METAL ION DEPOSITS IN BRAINS WITH
NEURODEGENERATIVE DISORDERS*
Ryuichi
Fukuyama §,
Akihiko
Nakayama¶,
Taizen
Nakase,
Hiroe
Toba ,
Teruo
Mukainaka**,
Hirofumi
Sakaguchi,
Takuya
Saiwaki§§,
Hiromu
Sakurai¶,
Mikio
Wada, and
Shinji
Fushiki
From the Department of Pathology and Applied
Neurobiology, Research Institute for Neurological Diseases and
Geriatrics, the ** Central Laboratory, and the
Department of Otolaryngology, Kyoto
Prefectural University of Medicine, 465 Kawaramachi Hirokoji,
Kamigyo-ku, Kyoto 602-8566 and the Departments of ¶ Analytical and
Bioinorganic Chemistry and Clinical Pharmacology, Kyoto
Pharmaceutical University, 5 Misasagi Nakauchi-cho,
Yamashina-ku, Kyoto 607-8414, Japan
From human neuroblastoma-derived SILA
cells we have established a  -0 cell line that is deficient in both
respiration and mitochondrial DNA. Lactate dehydrogenase activity,
lactate production, and growth in the medium without glucose indicate
that these cells shift from aerobic to anaerobic metabolism. Electron
microscopic observations revealed abnormal mitochondria with unique
cristae structures. Staining with MitoTracker dye showed that the
mitochondrial transmembrane potential was reduced by 30-40% from the
parent cell levels. These cells were markedly susceptible to
H2O2 and died apparently by a necrotic
mechanism, a process blocked by deferoxamine in the parent cells but
not -0 cells. Analysis by inductively coupled plasma-mass
spectrometry revealed an approximately 3-fold accumulation of iron in
the -0 cells at confluence (n = 4-6, three clones,
*p < 0.05). Iron and four other metals were all
elevated in the cells of one of the -0 clones and were similar to
control levels in the control cybrid cells, which were replenished with
normal mitochondrial DNA. Their sensitivity to
H2O2 was also similar to that of the parent
cells. These results indicate that a newly established neuronal related
-0 cell line is highly susceptible to active oxygen species and that
these toxicity effects appear to be related to an accumulation of
transition metals, which probably occurs through the respiratory impairment.
*
This research was supported in part by Grant-in-aid
H11-Chojyu-019 from the Japanese Foundation for Aging and Health and by the Ministry of Education, Culture, Sports, Science, and Technology of
Japan.The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
§
To whom correspondence should be addressed: Alzheimer Research
Laboratory, Dept. of Neurosciences, Case Western Reserve University School of Medicine, E504, 10900 Euclid Ave., Cleveland, OH
44106-4928. Tel.: 216-368-3435; Fax: 216-368-3079; E-mail:
rxf33@po.cwru.edu.
§§
Present address: Dept. of Cell Biology and Neuroscience, Graduate
School of Medicine, Osaka University, Yamadaoka 2-2, Suita, Osaka
565-0871, Japan.
Copyright © 2002 by The American Society for Biochemistry and Molecular Biology, Inc.
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Copyright © 2002 by the American Society for Biochemistry and Molecular Biology.
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