ALS-Linked Cu/Zn–SOD Mutation Increases Vulnerability of Motor Neurons to Excitotoxicity by a Mechanism Involving Increased Oxidative Stress and Perturbed Calcium Homeostasis

doi:10.1006/exnr.1999.7190

Experimental Neurology

Volume 160, Issue 1, November 1999, Pages 28-39

https://doi.org/10.1006/exnr.1999.7190 Get rights and content

Abstract

We employed a mouse model of ALS, in which overexpression of a familial ALS-linked Cu/Zn–SOD mutation leads to progressive MN loss and a clinical phenotype remarkably similar to that of human ALS patients, to directly test the excitotoxicity hypothesis of ALS. Under basal culture conditions, MNs in mixed spinal cord cultures from the Cu/Zn–SOD mutant mice exhibited enhanced oxyradical production, lipid peroxidation, increased intracellular calcium levels, decreased intramitochondrial calcium levels, and mitochondrial dysfunction. MNs from the Cu/Zn–SOD mutant mice exhibited greatly increased vulnerability to glutamate toxicity mediated by α-amino-3-hydroxy-5-methylisoxazole-4-propionate receptors. The increased vulnerability of MNs from Cu/Zn–SOD mutant mice to glutamate toxicity was associated with enhanced oxyradical production, sustained elevations of intracellular calcium levels, and mitochondrial dysfunction. Pretreatment of cultures with vitamin E, nitric oxide-suppressing agents, peroxynitrite scavengers, and estrogen protected MNs from Cu/Zn–SOD mutant mice against excitotoxicity. Excitotoxin-induced degeneration of spinal cord MNs in adult mice was more extensive in Cu/Zn–SOD mutant mice than in wild-type mice. The mitochondrial dysfunction associated with Cu/Zn–SOD mutations may play an important role in disturbing calcium homeostasis and increasing oxyradical production, thereby increasing the vulnerability of MNs to excitotoxicity.

References (64)

M.F. Beal
Mitochondrial dysfunction in neurodegenerative diseases
Biochim. Biophys. Acta
(1998)
P. Couratier et al.
Cell culture evidence for neuronal degeneration in amyotrophic lateral sclerosis being linked to glutamate AMPA/kainate receptors
Lancet
(1993)
L. Greensmith et al.
Blockade of N-methyl-d-aspartate receptors by MK-801 (dizocilpine maleate) rescues motoneurones in developing rats
Dev. Brain Res.
(1994)
M.P. Mattson et al.
Evidence for calcium-reducing and excitoprotective roles for the calcium binding protein (calbindin-D28k) in cultured hippocampal neurons
Neuron
(1991)
M.P. Mattson et al.
Growth factors prevent mitochondrial dysfunction, loss of calcium homeostasis, and cell injury, but not ATP depletion in hippocampal neurons deprived of glucose
Exp. Neurol.
(1993)
M.P. Mattson
Modification of ion homeostasis by lipid peroxidation: Roles in neuronal degeneration and adaptive plasticity
Trends Neurosci.
(1998)
J.M. McCord et al.
Superoxide dismutase: The first twenty years (1968–1988)
Free Rad. Biol. Med.
(1988)
S. Samarasinghe et al.
Distribution of the N-methyl-d-aspartate glutamate receptor subunit NR2A in control and amyotrophic lateral sclerosis spinal cord
Brain Res.
(1996)
S. Sasaki et al.
Ultrastructural study of synapses in the anterior horn neurons of patients with amyotrophic lateral sclerosis
Neurosci. Lett.
(1996)
P.J. Shaw et al.
The quantitative autoradiographic distribution of [³H]MK-801 binding sites in the normal human spinal cord
Brain Res.
(1991)

P.J. Shaw et al.

Non-NMDA receptors in motor neuron disease (MND): A quantitative autoradiographic study in spinal cord and motor cortex using [³H]CNQX and [³H]kainate

Brain Res.

(1994)

G.L. Squadrito et al.

Oxidative chemistry of nitric oxide: The roles of superoxide, peroxynitrite, and carbon dioxide

Free Rad. Biol. Med.

(1998)

S.A. Susin et al.

Mitochondria as regulators of apoptosis: Doubt no more

Biochim. Biophys. Acta

(1998)

R. Temkin et al.

Expression of glutamate receptor subunits in α-motoneurons

Mol. Brain Res.

(1997)

T.L. Williams et al.

An immunocytochemical study of the distribution of AMPA selective glutamate receptor subunits in the normal human motor system

Neuroscience

(1996)

P.C. Wong et al.

An adverse property of a familial ALS-linked SOD1 mutation causes motor neuron disease characterized by vacuolar degeneration of mitochondria

Neuron

(1995)

H.S. Yim et al.

A familial amyotrophic lateral sclerosis-associated A4V Cu,Zn-superoxide dismutase mutant has a lower Km for hydrogen peroxide: Correlation between clinical severity and the Km value

J. Biol. Chem.

(1997)

M. Yoshida et al.

Therapeutic effects of imidazolineoxyl N-oxide against endotoxin shock through its direct nitric oxide-scavenging activity

Biochem. Biophys. Res. Commun.

(1994)

M.F. Beal et al.

Increased 3-nitrotyrosine in both sporadic and familial amyotrophic lateral sclerosis

Ann. Neurol.

(1997)

G. Bensimon et al.

A controlled trial of riluzole in amyotrophic lateral sclerosis: ALS/Riluzole Study Group

N. Engl. J. Med.

(1994)

V.P. Bindokas et al.

Superoxide production in rat hippocampal neurons: Selective imaging with hydroethidine

J. Neurosci.

(1996)

S.G. Carriedo et al.

Motor neurons are selectively vulnerable to AMPA/kainate receptor-mediated injury in vitro

J. Neurosci.

(1996)

D.W. Choi

Calcium: Still center stage in hypoxic–ischemic neuronal death

Trends Neurosci.

(1996)

R.J. Ferrante et al.

Increased 3-nitrotyrosine and oxidative damage in mice with a human copper/zinc superoxide dismutase mutation

Ann. Neurol.

(1997)

R.J. Ferrante et al.

Evidence of increased oxidative damage in both sporadic and familial amyotrophic lateral sclerosis

J. Neurochem.

(1997)

Y. Goodman et al.

Ceramide protects hippocampal neurons against excitotoxic and oxidative insults, and amyloid β-peptide toxicity

J. Neurochem.

(1996)

Y. Goodman et al.

Estrogens attenuate and corticosterone exacerbates excitotoxicity, oxidative injury and amyloid β-peptide toxicity in hippocampal neurons

J. Neurochem.

(1996)

M.E. Gurney et al.

Motor neuron degeneration in mice that express a human Cu,Zn superoxide dismutase mutation

Science

(1994)

M.E. Gurney et al.

Pathogenic mechanisms in familial amyotrophic lateral sclerosis due to mutation of Cu, Zn superoxide dismutase

Pathol. Biol. (Paris)

(1996)

M.E. Gurney et al.

Benefit of vitamin E, riluzole, and gabapentin in a transgenic model of familial amyotrophic lateral sclerosis

Ann. Neurol.

(1996)

A. Hirano et al.

Fine structural observations of neurofilamentous changes in amyotrophic lateral sclerosis

J. Neuropathol. Exp. Neurol.

(1984)

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Regular ArticleALS-Linked Cu/Zn–SOD Mutation Increases Vulnerability of Motor Neurons to Excitotoxicity by a Mechanism Involving Increased Oxidative Stress and Perturbed Calcium Homeostasis

Abstract

Biochim. Biophys. Acta

Lancet

Dev. Brain Res.

Neuron

Exp. Neurol.

Trends Neurosci.

Free Rad. Biol. Med.

Brain Res.

Neurosci. Lett.

Brain Res.

Brain Res.

Free Rad. Biol. Med.

Biochim. Biophys. Acta

Mol. Brain Res.

Neuroscience

Neuron

J. Biol. Chem.

Biochem. Biophys. Res. Commun.

Increased 3-nitrotyrosine in both sporadic and familial amyotrophic lateral sclerosis

Ann. Neurol.

A controlled trial of riluzole in amyotrophic lateral sclerosis: ALS/Riluzole Study Group

N. Engl. J. Med.

Superoxide production in rat hippocampal neurons: Selective imaging with hydroethidine

J. Neurosci.

Motor neurons are selectively vulnerable to AMPA/kainate receptor-mediated injury in vitro

J. Neurosci.

Calcium: Still center stage in hypoxic–ischemic neuronal death

Trends Neurosci.

Increased 3-nitrotyrosine and oxidative damage in mice with a human copper/zinc superoxide dismutase mutation

Ann. Neurol.

Evidence of increased oxidative damage in both sporadic and familial amyotrophic lateral sclerosis

J. Neurochem.

Ceramide protects hippocampal neurons against excitotoxic and oxidative insults, and amyloid β-peptide toxicity

J. Neurochem.

Estrogens attenuate and corticosterone exacerbates excitotoxicity, oxidative injury and amyloid β-peptide toxicity in hippocampal neurons

J. Neurochem.

Motor neuron degeneration in mice that express a human Cu,Zn superoxide dismutase mutation

Science

Pathogenic mechanisms in familial amyotrophic lateral sclerosis due to mutation of Cu, Zn superoxide dismutase

Pathol. Biol. (Paris)

Benefit of vitamin E, riluzole, and gabapentin in a transgenic model of familial amyotrophic lateral sclerosis

Ann. Neurol.

Fine structural observations of neurofilamentous changes in amyotrophic lateral sclerosis

J. Neuropathol. Exp. Neurol.

Regular Article
ALS-Linked Cu/Zn–SOD Mutation Increases Vulnerability of Motor Neurons to Excitotoxicity by a Mechanism Involving Increased Oxidative Stress and Perturbed Calcium Homeostasis