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The Journal of Neuroscience, May 15, 2001, 21(10):3369-3374
Neuron-Specific Expression of Mutant Superoxide Dismutase 1 in
Transgenic Mice Does Not Lead to Motor Impairment
Albéna
Pramatarova,
Janet
Laganière,
Julie
Roussel,
Katéri
Brisebois, and
Guy A.
Rouleau
Centre for Research in Neuroscience, McGill University, and the
Montreal General Hospital Research Institute, Montreal, Quebec, H3G
1A4, Canada
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ABSTRACT |
Mutations were identified in the Cu/Zn superoxide dismutase gene
(SOD1) in ~15% of patients with familial amyotrophic lateral sclerosis. Transgenic animals expressing mutant SOD1 in all tissues develop an ALS-like phenotype. To determine whether neuron-specific expression of mutant SOD1 is sufficient to produce such a phenotype, we
generated transgenic animals carrying the G37R mutation that is
associated with the familial form of ALS (FALS), which is driven by the
neurofilament light chain promoter. The transgenic animals express high
levels of the human SOD1 protein in neuronal tissues, especially in the
large motor neurons of the spinal cord, but they show no apparent motor
deficit at up to 1.5 years of age. Our animal model suggests that
neuron-specific expression of ALS-associated mutant human SOD1 may not
be sufficient for the development of the disease in mice.
Key words:
amyotrophic lateral sclerosis; superoxide dismutase; mutant SOD1; transgenic mice; neuronal expression; cell death; neurofilament promoter
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INTRODUCTION |
Amyotrophic lateral sclerosis (ALS)
is a late onset neurodegenerative disorder characterized by the death
of large motor neurons in the cerebral cortex and spinal cord (Tandan
and Bradley, 1985 ). The familial form of ALS (FALS) accounts for
~10% of cases and usually is transmitted as an autosomal
dominant trait (Mulder et al., 1986). Mutations in Cu/Zn superoxide
dismutase gene (SOD1), a ubiquitously expressed and highly conserved
metalloenzyme involved in the detoxification of free radicals, are
responsible for ~15% of FALS (Rosen et al., 1993; Fridovich, 1995;
Pramatarova et al., 1995).
In vitro studies showed that some mutants had a reduced
specific activity whereas others retained full activity (Borchelt et
al., 1994). The wide variety of enzymatic activities and the absence of
motor dysfunction in transgenic SOD1 null mice make loss of function an
unlikely pathogenic mechanism of FALS and suggest that mutant SOD1
toxicity arises from a gain rather than from a loss of function (Reaume
et al., 1996).
Consistent with this model, transgenic mice overexpressing
FALS-associated SOD1 mutants (G93A, G37R, and G85R) developed a rapidly
progressive disease with lower motor neuron degeneration that is
reminiscent of human ALS (Gurney et al., 1994; Dal Canto and Gurney,
1995; Wong et al., 1995; Mourelatos et al., 1996; Bruijn et al., 1997).
In G37R or G93A animals high levels of mutant SOD1 expression were
required to develop the disease, and the total SOD1 enzyme activity was
increased 4- to 14-fold. In these models the age of onset of disease
and neuronal populations that were affected depended on the levels of
mutant enzyme. On the other hand, transgenic animals expressing the
G85R mutation developed a rapidly progressing fatal disease, astrocytic
inclusions that stained positive for SOD1, and ubiquitin as well
as SOD1-positive aggregates in motor neurons, despite modest levels of
expression (Bruijn et al., 1997).
Although these animal models implicate mutant SOD1 in the development
of motor neuron degeneration, many questions remain, particularly with
respect to the mechanism of disease pathogenesis. One critical question
is whether cellular damage arises from direct motor neuron toxicity or
whether indirect toxic effects, for example via the astrocytes as
suggested by the G85R transgenics, contribute to neuronal cell death. A
recent study showed that expression of mutant mouse G86R SOD1 cDNA in
the astrocytes of transgenic mice was not sufficient to cause an
ALS-like motor deficit (Gong et al., 2000).
To test whether motor neuron expression of mutant SOD1 is sufficient
for disease development, we generated transgenic animals carrying a
human SOD1 cDNA with the G37R FALS-associated mutation that is driven
by the neurofilament light chain promoter. We found that, in contrast
to observations in transgenic lines overexpressing FALS-associated SOD1
mutations in a constitutive manner, neuron-specific expression does not
cause significant motor neuron cell death.
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MATERIALS AND METHODS |
Construction of transgenic mice expressing wild-type and
mutant G37R human SOD1. Total human RNA was extracted from
lymphoblasts by homogenization in guanidinium thiocyanate and
ultracentrifugation through a cesium chloride cushion for 20 hr at
35,000 rpm. Then the RNA pellet was washed, resuspended in water, and
kept at  20°C (Chirgwin et al., 1979). RT-PCR was performed on 2 µg of RNA, using the primers SOD1A, 5'-AAG TCG ACA AGC TTT GCG TCG
TAG TCT CCT GCA-3', and SOD1B, 5'-GCC CTC GAG AAG CTT TTT TTT AAG ATT ACA GTG-3', which introduce a HinDIII restriction site. The
cDNA was cloned and sequenced with a Sequenase kit (Amersham Life
Science, Arlington Heights, IL). We introduced the mutation G37R by PCR site-directed mutagenesis (Tomic et al., 1990). The vector carrying the
neurofilament light chain promoter (pGCHNFL) was generously provided by
Dr. J. P. Julien (McGill University, Montreal, Canada).
Transgenic animals were generated via standard microinjection
techniques that used hybrid mouse embryos (C57BL/6J × C3H/HeJ) (Brinster et al., 1981). To test for transgene transmission, we isolated DNA from mouse tail; we performed Southern blotting by using the human SOD1 cDNA as probe. The animals were housed in a
pathogen-free environment, were checked on a regular basis for microbial infections, and also were examined for motor dysfunction (general motility and activity, grooming, holding by the tail, hanging
to a rod; Collard and Julien, 1995).
Expression analysis. Mice (6-7 months of age) were
killed by cervical dislocation; the dissected tissues (brain,
spinal cord, heart, liver, kidney, spleen, muscle, and testis) were
frozen rapidly in liquid nitrogen and then stored at 80°C until
used for RNA or protein extraction.
Multiple tissues RT-PCR was performed on 1 µg of total DNase-treated
RNA, with an oligo-dT primer, using the kit from PerkinElmer Life
Sciences (Norwalk, CT). PCR was performed for 35 cycles at 94°C for 1 min, 60°C for 30 sec, and 72°C for 45 sec, using
primers SOD1A and SOD1B, and amplifying specifically the human SOD1
cDNA. RT-PCR was performed also, in parallel, for 35 cycles of 94°C for 1 min, 62°C for 30 sec, and 72°C for 45 sec, using primers amplifying the GAPDH cDNA (5'-CCT TTC ATT GAC CTC AAC TAC ATG G-3' and
5'-AGT CTT CTG GGT GGC AGT GAT GG-3') (Hyman Friedman et al.,
1996).
Protein was extracted by homogenization of the tissues in lysis buffer
(1% NP-40 and 1 mM PMSF in PBS), and centrifugation for 20 min at 13,000 rpm at 10°C. The soluble protein was quantified by the
Lowry method and diluted in loading buffer (15% glycerol, 5% SDS, 80 mM Tris-HCl, pH 6.8, 5%  -mercapto-ethanol, and 0.01% bromophenol blue). Each sample (100 µg) was run on a 12% SDS/glycine polyacrylamide gel according to Laemmli and then transferred
electrophoretically on a nitrocellulose membrane (Schleicher & Schuell,
Dassel, Germany). Human SOD1 (20-40 ng) was used as a positive control
(Sigma, St. Louis, MO). The blot was blocked in 2% nonfat milk/0.1%
Tween 20 in PBS and probed with a sheep anti-human SOD1 antibody
diluted 1:500 in the blocking buffer (Biodesign, Kennebunkport, ME).
The immunodetection was performed with an HRP-labeled secondary
antibody, and the detection was done with the Renaissance kit
(PerkinElmer Life Sciences). For the protein quantification serial
dilutions of each sample (10-150 µg) as well as diluted hSOD1
(0.6-40 ng) were run on 12% SDS/glycine polyacrylamide gels,
transferred on nitrocellulose, probed, and developed as previously
described; densitometry analysis was performed with the Is-1000 Digital
Imaging System from Alpha Innotech (San Leandro, CA).
Generation and characterization of the LAP2-10AP anti-human SOD1
antibody. Antibodies were raised against a region of the human
SOD1 protein (hSOD1) spanning amino acids 18-37, which displays low
homology with the mouse SOD1 protein (mSOD1; 50% identity). A
synthetic peptide (IINFEQKESNGPVKVWGSIK+C) was cross-linked to keyhole
limpet hemocyanin (KLH), emulsified in Freund's adjuvant, and used to
immunize rabbits (1 mg in complete Freund's adjuvant for the first
injection, followed every 4-6 weeks by 0.5 mg in incomplete Freund's
adjuvant). Rabbit serum was affinity-purified on a column, and aliquots
were kept frozen at 20°C. The affinity-purified hSOD1 antibody
(LAP2-10AP) was characterized by immunoblotting. Human SOD1 (Sigma) and
protein extracts from spinal cord of transgenic animals overexpressing
the human SOD1 as well as of nontransgenic animals were run on a 12%
SDS/glycine polyacrylamide gel and electrotransferred on
nitrocellulose. The blots were blocked in 2% nonfat milk/0.1% Tween
20 in PBS and then incubated with either preimmune serum (LAP2-1) or
hSOD1 antibody or with peptide-preabsorbed hSOD1 antibody diluted 1:500
in the blocking buffer. The immunodetection was done by incubating the
blots with a biotinylated goat anti-rabbit antibody, followed by
incubation with streptavidin-HRP according to the instructions from
the manufacturer; final detection was made with the Renaissance kit
(PerkinElmer Life Sciences).
Immunocytochemical analysis and morphometry. Mice (8 months
old) were anesthetized with an intraperitoneal injection of 4% chloral
hydrate and perfused intracardially with PBS, followed by 4%
paraformaldehyde, pH 7.4, in 0.1 M sodium phosphate buffer. Spinal cords were removed, post-fixed for 1 hr, and then frozen in
Tissue-Tek OCT embedding compound (Sakura Finetek, Torrance, CA).
Immunocytochemistry was performed on 30-µm-thick floating sections.
Sections were quenched first in 0.3%
H2O2 for 30 min, permeabilized in 1% Tween 20 for 10 min, blocked in 3% normal goat
serum for 3 hr, and then incubated in the primary hSOD1 antibody (LAP2-10AP) for 16 hr at 4°C. The immunoreactivity was visualized with diaminobenzidine after amplification of the signal with the TSA-indirect reagent (PerkinElmer Life Sciences) and Vectastain ABC
Elite (Vector Laboratories, Burlingame, CA).
To determine the percentage of hSOD1-positive cells in the spinal cords
that were analyzed, we photographed multiple sections, circled the
hSOD1-positive cells, and determined their relative surface area with
an Image 1 Analyzer (Universal Imaging, West Chester, PA).
Motor function measurements. Motor capacity was assessed by
measuring the grip strength of the animals. The data were
analyzed by ANOVA.
Microscopic analysis and quantification of motor neuron
degeneration. Mice (11-12 months old) were anesthetized with an
intraperitoneal injection of 4% chloral hydrate and perfused with PBS,
followed by 3% glutaraldehyde, pH 7.4, in 0.1 M sodium
phosphate buffer. Ventral roots from levels L4 and L5 were removed,
post-fixed for 16 hr, incubated in 1% osmium tetroxide, dehydrated in
a graded series of ethanol, and processed for embedding in Epon. Thick sections (1 µm) were stained with 1% toluidine blue and
photographed. The axons in cross sections of the ventral roots were
circled on the photographs, and the sizes were determined with an Image 1 Analyzer (Universal Imaging). The measurements were grouped by size
and analyzed by an ANOVA test.
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RESULTS |
Transgenic lines expressing wild-type and FALS-associated G37R
mutant human SOD1
Transgenic mice were created by using a 10 kb construct carrying a
human SOD1 cDNA that is driven by a neurofilament light chain promoter
system, which contains some genomic regulatory elements destined to
ensure adequate levels and localization of expression (Fig.
1,1) (Charron et al., 1995).
Two different constructs were used, one containing a normal human SOD1
cDNA and one carrying a FALS-associated mutation G37R (GGA AGA)
(Pramatarova et al., 1995). Founder animals were identified by Southern
blotting of tail DNA; three lines were established, one carrying the
normal cDNA (SOD1-4305) and two lines carrying the mutant cDNA
(G37R-3156 and G37R-4012).
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Figure 1.
1, Diagram of the construct used to
generate the transgenic animals. The pGCHNFL vector contains the
promoter region, some enhancing intronic sequences, and the
3'-untranslated region (3'-UTR) of the neurofilament
light chain gene. 2, RT-PCR and Western blots show
expression of the human transgene in nervous tissue both at the mRNA
and protein levels. 2A, Multiple tissue RT-PCR showing
expression of the human SOD1 cDNA. 2B, Same as
2A but using primers amplifying the GAPDH cDNA.
2C, Western blots probed with an anti-SOD1 antibody
recognizing both human and mouse proteins, showing expression of the
hSOD1 in nervous tissue exclusively. mSOD1, Mouse SOD1;
hSOD1, human SOD1.
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For each line, RNA was extracted from brain, spinal cord, heart, liver,
kidney, spleen, muscle, and testis and treated with DNase to avoid DNA
contamination and false positives. RT-PCR showed expression of the
transgene in brain and spinal cord (Fig. 1,2A), as expected.
The signal detected in certain other tissues like the heart, kidney,
and spleen may result from transcription caused by the integration site
of the transgene.
Expression of the transgene at the protein level was assessed by
Western blotting of total soluble protein from brain, spinal cord,
heart, liver, kidney, spleen, muscle, and testis. The blots were probed
with a polyclonal antibody (Biodesign) recognizing both human (hSOD1)
and mouse (mSOD1) proteins. All three lines show expression of the
hSOD1 exclusively in brain and spinal cord, but not in any other tissue
(Fig. 1,2C). The hSOD1 concentration is highest in the
spinal cord where the large neurons are concentrated.
To quantify the amount of hSOD1 in the spinal cords, we ran serial
dilutions of each brain and spinal cord extract as well as known
amounts of commercial hSOD1 protein. The amount of human protein was
estimated by densitometry (Table 1). For
the SOD1-4305 line (normal SOD1 control) the amount of protein in the
total brain extract was too small to be quantified reliably by our
method.
Localization and quantification of transgene expression: high
levels of human SOD1 expressed in large motor neurons
To determine the cell-specific expression of hSOD1 in the
transgenic mice, we raised antisera to the human SOD1 protein by using
a peptide displaying low homology with the mouse SOD1 protein; i.e., 10 of 20 amino acids are human-specific. Serum was harvested and
affinity-purified against the peptide that was used for the immunization. The preimmune serum and the affinity-purified hSOD1 antibody were tested on immunoblots. The hSOD1 antibody detects the
hSOD1 (from total cell extracts and from the commercially purified) and
does not cross-react with the mouse SOD1 (Fig.
2). Preabsorption of the hSOD1 antibody
with the peptide abolishes the hSOD1 reactivity.
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Figure 2.
Characterization of the human SOD1 antibody.
A, Western blot probed with preimmune serum.
B, Western blot probed with the human SOD1 antibody,
which specifically recognizes the hSOD1, but not the mSOD1, protein.
C, Western blot probed with peptide-preabsorbed human
SOD1 antibody. h/mSOD1, Protein extract from spinal cord
of transgenic animal overexpressing hSOD1; mSOD1,
protein extract from spinal cord of nontransgenic animal;
hSOD1, human SOD1 (Sigma).
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Immunocytochemistry was performed on samples from each transgenic line
as well as on normal nontransgenic animals. Using the human-specific
SOD1 antibody, we were able to show that the hSOD1 protein is expressed
specifically in a subset of large neurons concentrated in the ventral
horns of the spinal cords of transgenic animals (Fig.
3).
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Figure 3.
Human SOD1 is expressed in large neurons in the
ventral horn of the spinal cord in transgenic animals. A,
B, Normal nontransgenic animal. C, D, Transgenic
line G37R-3156. E, F, Transgenic line G37R-4012.
G, H, Transgenic line SOD1-4305. Scale bars: A,
C, E, G, 200 µm; B, 100 µm; D, F,
H, 50 µm.
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Because only a proportion of cells in the spinal cord expresses hSOD1,
we undertook to determine specifically the amount of hSOD1 per motor
neuron. We morphometrically estimated what percentage of the total
surface area of the spinal cord is occupied by hSOD1-positive cells.
Depending on the different lines, between ~2 and 5% of the surface
area of the spinal cord stained positively for hSOD1 (Table
2).
Total hSOD1 in spinal cords was quantified by Western blotting (see
Table 1), and a "corrected" concentration of hSOD1 in those
specific hSOD1-positive cells in the spinal cords of the transgenic
animals was determined (Table 2), ranging from 2.3 to 8.1 ng/µg of
total protein. These amounts of human SOD1 are equivalent to a ratio of
4.3 of human-to-mouse SOD1 in the highest expressing line. This level
is comparable with the values reported by other groups in transgenic
animals expressing the G37R or G93A mutations in a constitutive manner
and that show an ALS-like phenotype (Table
3) (Gurney et al., 1994; Wong et al.,
1995).
Absence of motor degeneration in G37R transgenic animals
The survival of the transgenic animals was similar in all
lines, with 97% of the mice from line G37R-3156 (n = 41) living for >1 year, 96% of line G37R-4012 (n = 46) and 82% of line SOD1-4305 (n = 11). Furthermore,
85% of lines G37R-3156 and G37R-4012 and 73% of the mice from line
SOD1-4305 survived 1.5 years or older.
The transgenic animals were examined on a weekly basis for
general health (grooming, general activity) but showed no apparent motor dysfunction at up to 1.5 years of age. More specifically, there
was no significant difference in performance between the transgenic and
nontransgenic animals subjected to grip strength tests (Fig.
4). We have compared toluidine
blue-stained spinal cord sections from 1-year-old animals and found no
significant differences among the different lines (data not shown).
Because our animals did not show any gross motor dysfunction, they also were tested for axonal degeneration, often an early sign of neuronal degeneration. One well documented approach is to quantify the size and
number of the motor neuron axons in the ventral root of the spinal cord
of the mice. Animals aged between 11 and 12 months ("middle" aged)
were used to avoid confusion with normal age-related neurodegeneration.
Both L4 and L5 ventral roots were examined and showed no apparent
degeneration in our transgenic animals (Fig.
5, 1). The size and number of
axons were determined, and no significant differences were seen
between the transgenic and nontransgenic animals (Fig.
5B).
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Figure 4.
Limb grip strength measurement. There is no
significant difference in the performance of transgenic versus
nontransgenic animals.
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Figure 5.
Size of motor neurons in L4 and L5 ventral roots.
1, Photographs of ventral root axons of 1-year-old
normal and transgenic animals. 1A, Nontransgenic animal.
1B, Line SOD1-4305. 1C, Line G37R-3156.
1D, Line G37R-4012. Scale bar, 100 µm.
2, Axon counts in L4 and L5 ventral roots of 1-year-old
normal and transgenic animals. 1, 2, No significant
differences are seen between the normal and transgenic animals.
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DISCUSSION |
The neurofilament light chain promoter allowed us to express high
levels of human SOD1 specifically in large motor neurons of the spinal
cord of transgenic animals. Surprisingly, these animals did not develop
a neurodegenerative disease. Other groups have shown previously that
the same G37R mutant, when expressed ubiquitously under the SOD1
promoter in mice, leads to a disorder characterized by weakness of the
limbs, muscle wasting, and paralysis, with pathological changes
reminiscent of human ALS (Wong et al., 1995). At the ultrastructural
level motor neurons in the ventral horn of the spinal cord as well as
in the brainstem were degenerating with cytoplasmic vacuoles in
dendrites, proximal axons, and perikarya, containing degenerating and
swollen mitochondria. The age at onset of disease in these mice
appeared to be dependent on the amount of mutant protein, often
expressed as the human-to-mouse SOD1 ratio, and occurred as early as
3.5 months of age for the high expressors, with ratios of 12:1 in the
spinal cord; most importantly, for lines expressing levels similar to
our transgenics (4-5:1), the disease developed at 6-8 months of age,
with a significant loss of motor neurons by 20 weeks of age. Our model,
which expressed similar high levels of human mutant protein (4.3:1
ratio) in motor neurons, the cells most affected in humans and mice,
appeared healthy at >18 months of age. The absence of phenotype in our animals expressing the mutant protein in only a subset of neurons suggests that the localization of the transgene expression is important
for neurodegeneration.
There are two possible explanations for the absence of phenotype in our
animals. The first is that insufficient levels of transgene expression
were achieved. It is well established that the levels of mutant
transgene expression are crucial for determining the age at onset of
the disease in transgenic animals. This raises the concern that our
animals might not be expressing high enough levels of mutant protein.
Although we estimated that the mutant human SOD1 levels in individual
motor neurons of the spinal cord of our animals were comparable with
the levels achieved in the other animal models, which did develop motor
dysfunction, our methods of dosage may have overestimated the human
SOD1 levels, or the published ratios may have been underestimated. When
the SOD1 gene is regulated by its own promoter, the protein is
expressed at high levels throughout the entire life of the animals,
whereas the NFL promoter is downregulated slightly later in life
(Kuchel et al., 1997). The damage caused by mutant SOD1 is thought to be cumulative, but it is probably also important to have high levels of
the defective protein later in life when the damage starts to be
visible. In our model this should not be the cause of the absence of
phenotype because the G37R mutant, when expressed at a 4-5:1 of
human-to-mouse ratio, induces the disease very early in the life of
transgenic animals, long before the NFL age-related downregulation
occurs. Therefore, our mice should have achieved the desired levels of
expression at the appropriate time for the disease to develop.
Another important point to consider is that there might be a threshold
of human mutant protein for the disease to appear, which varies
according to the mutation. For example, there seems to be a lower
threshold for the G85R mutant (0.2-1:1 human-to-mouse ratio) and a
higher one for G37R mutant (4-5:1 ratio). This threshold also could
differ according to the localization of the expression (ubiquitous vs
neuronal), which might result in the need to reach a higher threshold
for neuron-specific expression to provoke the disease in a similar time frame.
The second explanation for the absence of phenotype in our mice is that
the presence of the mutant protein in motor neurons might not be
sufficient to lead to the development of a neurodegenerative disease.
Widespread expression of the mutant SOD1 might be necessary for the
development of the disease. One possible pathogenic model that requires
extra-neuronal expression of the mutant protein is suggested by the
G85R mouse model. In these animals the first signs of damage were shown
in the astrocytes (Bruijn et al., 1997). In parallel with the neuronal
loss and astrocytic inclusions, at end stage the G85R mice showed a
50% decrease in the astrocyte-specific glutamate transporter GLT-1,
which is the major glutamate transporter in the spinal cord. This model
suggests that astrocyte expression with loss of astrocytic function and
perhaps loss of glutamate uptake may be important in disease
development. The recent report by Gong et al. (2000), who found that
astrocytic expression of mutant SOD1 leads to astrocytic changes,
supports the hypothesis that these cells may be involved in the motor
neuron cell death that is seen in this model of ALS.
The importance of glutamate toxicity was suggested by experiments of
nonselective blockage of glutamate transporters, which lead to
pathological abnormalities reminiscent of neurodegenerative diseases
with cellular swellings and vacuolation of mitochondria and endoplasmic
reticulum (Choi, 1988; Ikonomidou et al., 1996; Rothstein et al.,
1996). A selective decrease in glutamate transporters has been observed
in multiple animal models of motor neuron disease (Mnd, G85R). At the
ultrastructural level the pathology seen in the animal models of ALS
(G37R, G93A) is very similar to the neurodegeneration induced by
excitotoxins (agonists of glutamate transporters) (Ikonomidou et al.,
1996). More significantly, a decrease in glutamate transporter GLT-1
and an increase in extracellular glutamate in the CSF were shown in ALS
patients (Rothstein et al., 1990, 1992, 1995). These data suggest that
glutamate, via the astrocytes, might induce/propagate/aggravate the
motor neuron-specific degeneration seen in FALS and in the animal
models of the disease. This mechanism could explain our observation
that transgenic animals expressing high levels of mutant human SOD1 in
motor neurons do not develop the disease. Additional experiments need
to be performed to test this hypothesis.
We have generated transgenic animals carrying a human SOD1 gene with
the G37R mutation associated with FALS and an ALS-like phenotype in
transgenic mice. The neurofilament light chain promoter was used
successfully to express specifically the mutant protein in neurons, the
cells that are depleted in ALS patients and in animal models of ALS.
Our transgenic animals express high levels of the mutant human SOD1
protein in nervous tissue, especially in the large neurons of the
spinal cord, which degenerate in ALS but remain healthy at up to 1.5 years of age, suggesting that mutant SOD1 expression in other cell
types may contribute to motor neuron loss that is caused by mutant SOD1
toxicity in mice.
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FOOTNOTES |
Received Sept. 6, 2000; revised Feb. 26, 2001; accepted March 7, 2001.
This work was supported by the Muscular Dystrophy Association, the
Muscular Dystrophy Association of Canada, the Medical Research Council
of Canada, and the Fonds de Recherche en Santé du Québec. Scholarships to A.P. were provided by the Fonds pour la Formation de
Chercheurs et l'Aide à la Recherche and the McGill University Faculty of Medicine. The production of transgenic mice was supported by
the Canadian Network of Centres of Excellence. We thank Dr. Jean Pierre
Julien for providing the pGCHNFL vector and Dr. Philip Wong for the
G37R29 transgenic line that was used as a positive control in Figure 2.
We are especially grateful to Dr. Diane Merry for helping with the grip
strength measurements.
Correspondence should be addressed to Guy A. Rouleau, Centre for
Research in Neuroscience, Montreal General Hospital, 1650 Cedar Avenue,
Montreal, Quebec, H3G 1A4, Canada. E-mail:
mi32{at}musica.mcgill.ca.
A. Pramatarova's present address: Neurogenetics Branch,
National Institute of Neurological Disorders and Stroke, National Institutes of Health, Bethesda, MD 20892.
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REFERENCES |
-
Borchelt DR,
Lee MK,
Slunt HS,
Guarnieri M,
Xu ZS,
Wong PC,
Brown Jr RH,
Price DL,
Sisodia SS,
Cleveland DW
(1994)
Superoxide dismutase 1 with mutations linked to familial amyotrophic lateral sclerosis possesses significant activity.
Proc Natl Acad Sci USA
91:8292-8296[Abstract/Free Full Text].
-
Brinster RL,
Chen HY,
Trumbauer M,
Senear AW,
Warren R,
Palmiter RD
(1981)
Somatic expression of herpes thymidine kinase in mice following injection of a fusion gene into eggs.
Cell
27:223-231[Web of Science][Medline].
-
Bruijn LI,
Becher MW,
Lee MK,
Anderson KL,
Jenkins NA,
Copeland NG,
Sisodia SS,
Rothstein JD,
Borchelt DR,
Price DL,
Cleveland DW
(1997)
ALS-linked SOD1 mutant G85R mediates damage to astrocytes and promotes rapidly progressive disease with SOD1-containing inclusions.
Neuron
18:327-338[Web of Science][Medline].
-
Ceballos-Picot I,
Nicole A,
Briand P,
Grimber G,
Delacourte A,
Defossez A,
Javoy-Agid F,
Lafon M,
Blouin JL,
Sinet PM
(1991)
Neuronal-specific expression of human copper/zinc superoxide dismutase gene in transgenic mice: animal model of gene dosage effects in Down's syndrome.
Brain Res
552:198-214[Web of Science][Medline].
-
Charron G,
Guy LG,
Bazinet M,
Julien JP
(1995)
Multiple neuron-specific enhancers in the gene coding for the human neurofilament light chain.
J Biol Chem
270:30604-30610[Abstract/Free Full Text].
-
Chirgwin JM,
Przybyla AE,
MacDonald RJ,
Rutter WJ
(1979)
Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease.
Biochemistry
18:5294-5299[Medline].
-
Choi DW
(1988)
Glutamate neurotoxicity and diseases of the nervous system.
Neuron
1:623-634[Web of Science][Medline].
-
Collard JF,
Julien JP
(1995)
A simple test to monitor the motor dysfunction in a transgenic mouse model of amyotrophic lateral sclerosis.
J Psychiatry Neurosci
20:80-86[Medline].
-
Dal Canto MC,
Gurney ME
(1995)
Neuropathological changes in two lines of mice carrying a transgene for mutant human Cu/Zn SOD, and in mice overexpressing wild-type human SOD: a model of familial amyotrophic lateral sclerosis (FALS).
Brain Res
676:25-40[Web of Science][Medline].
-
Epstein CJ,
Avraham KB,
Lovett M,
Smith S,
Elroy-Stein O,
Rotman G,
Bry C,
Groner Y
(1987)
Transgenic mice with increased Cu/Zn superoxide dismutase activity: animal model of dosage effects in Down syndrome.
Proc Natl Acad Sci USA
84:8044-8048[Abstract/Free Full Text].
-
Fridovich I
(1995)
Superoxide radical and superoxide dismutases.
Annu Rev Biochem
64:97-112[Web of Science][Medline].
-
Gong YH,
Parsadanian AS,
Andreeva A,
Snider WD,
Elliott JL
(2000)
Restricted expression of G86R Cu/Zn superoxide dismutase in astrocytes results in astrocytosis but does not cause motoneuron degeneration.
J Neurosci
20:660-665[Abstract/Free Full Text].
-
Gurney ME,
Pu H,
Chiu AY,
Dal Canto MC,
Polchow CY,
Alexander DD,
Caliendo J,
Hentati A,
Kwon YW,
Deng HX
(1994)
Motor neuron degeneration in mice that express a human Cu/Zn superoxide dismutase mutation.
Science
264:1772-1775[Abstract/Free Full Text].
-
Hyman Friedman HC,
Jelsma TN,
Bray GM,
Aguayo AJ
(1996)
A distinct pattern of trophic factor expression in myelin-deficient nerves of Trembler mice: implications for trophic support by Schwann cells.
J Neurosci
16:5344-5350[Abstract/Free Full Text].
-
Ikonomidou C,
Qin Qin Y,
Labruyere J,
Olney JW
(1996)
Motor neuron degeneration induced by excitotoxin agonists has features in common with those seen in the SOD-1 transgenic mouse model of amyotrophic lateral sclerosis.
J Neuropathol Exp Neurol
55:211-224[Web of Science][Medline].
-
Kuchel GA,
Poon T,
Irshad K,
Richard C,
Julien JP,
Cowen T
(1997)
Decreased neurofilament gene expression is an index of selective axonal hypotrophy in aging.
NeuroReport
8:799-805[Web of Science][Medline].
-
Mourelatos Z,
Gonatas NK,
Stieber A,
Gurney ME,
Dal Canto MC
(1996)
The Golgi apparatus of spinal cord motor neurons in transgenic mice expressing mutant Cu/Zn superoxide dismutase becomes fragmented in early, preclinical stages of the disease.
Proc Natl Acad Sci USA
93:5472-5477[Abstract/Free Full Text].
-
Mulder DW,
Kurland LT,
Offord KP,
Beard CM
(1986)
Familial adult motor neuron disease: amyotrophic lateral sclerosis.
Neurology
36:511-517[Abstract].
-
Pramatarova A,
Figlewicz DA,
Krizus A,
Han FY,
Ceballos-Picot I,
Nicole A,
Dib M,
Meininger V,
Brown RH,
Rouleau GA
(1995)
Identification of new mutations in the Cu/Zn superoxide dismutase gene of patients with familial amyotrophic lateral sclerosis.
Am J Hum Genet
56:592-596[Web of Science][Medline].
-
Reaume AG,
Elliott JL,
Hoffman EK,
Kowall NW,
Ferrante RJ,
Siwek DF,
Wilcox HM,
Flood DG,
Beal MF,
Brown Jr RH,
Scott RW,
Snider WD
(1996)
Motor neurons in Cu/Zn superoxide dismutase-deficient mice develop normally but exhibit enhanced cell death after axonal injury.
Nat Genet
13:43-47[Web of Science][Medline].
-
Ripps ME,
Huntley GW,
Hof PR,
Morrison JH,
Gordon JW
(1995)
Transgenic mice expressing an altered murine superoxide dismutase gene provide an animal model of amyotrophic lateral sclerosis.
Proc Natl Acad Sci USA
92:689-693[Abstract/Free Full Text].
-
Rosen DR,
Siddique T,
Patterson D,
Figlewicz DA,
Sapp P,
Hentati A,
Donaldson D,
Goto J,
O'Regan JP,
Deng HX,
Rahmani Z,
Krizus A,
McKenna-Yasek D,
Cayabyab A,
Gaston SM,
Berger R,
Tanzy RE,
Halperin JJ,
Herzfeldt B,
Van den Bergh R,
Hung WY,
Bird T,
Deng G,
Mulder DW,
Smyth C,
Laing NG,
Soriano E,
Pericak-Vance MA,
Hanes H,
Rouleau GA,
Gusella JS,
Horvitz HR,
Brown RH
(1993)
Mutations in Cu/Zn superoxide dismutase gene are associated with familial amyotrophic lateral sclerosis.
Nature
362:59-62[Medline].
-
Rothstein JD,
Tsai G,
Kuncl RW,
Clawson L,
Cornblath DR,
Drachman DB,
Pestronk A,
Stauch BL,
Coyle JT
(1990)
Abnormal excitatory amino acid metabolism in amyotrophic lateral sclerosis.
Ann Neurol
28:18-25[Web of Science][Medline].
-
Rothstein JD,
Martin LJ,
Kuncl RW
(1992)
Decreased glutamate transport by the brain and spinal cord in amyotrophic lateral sclerosis.
N Engl J Med
326:1464-1468[Abstract].
-
Rothstein JD,
Van Kammen M,
Levey AI,
Martin LJ,
Kuncl RW
(1995)
Selective loss of glial glutamate transporter GLT-1 in amyotrophic lateral sclerosis.
Ann Neurol
38:73-84[Web of Science][Medline].
-
Rothstein JD,
Dykes-Hoberg M,
Pardo CA,
Bristol LA,
Jin L,
Kuncl RW,
Kanai Y,
Hediger MA,
Wang Y,
Schielke JP,
Welty DF
(1996)
Knock-out of glutamate transporters reveals a major role for astroglial transport in excitotoxicity and clearance of glutamate.
Neuron
16:675-686[Web of Science][Medline].
-
Tandan R,
Bradley WG
(1985)
Amyotrophic lateral sclerosis. Part 1. Clinical features, pathology, and ethical issues in management.
Ann Neurol
18:271-280[Web of Science][Medline].
-
Tomic M,
Sunjevaric I,
Savtchenko ES,
Blumenberg M
(1990)
A rapid and simple method for introducing specific mutations into any position of DNA leaving all other positions unaltered.
Nucleic Acids Res
18:1656[Free Full Text].
-
Wong PC,
Pardo CA,
Borchelt DR,
Lee MK,
Copeland NG,
Jenkins NA,
Sisodia SS,
Cleveland DW,
Price DL
(1995)
An adverse property of a familial ALS-linked SOD1 mutation causes motor neuron disease characterized by vacuolar degeneration of mitochondria.
Neuron
14:1105-1116[Web of Science][Medline].
Copyright © 2001 Society for Neuroscience 0270-6474/01/21103369-06$05.00/0
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|
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[PDF]
|
|
|
|
|
|
|
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106(7):
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[Full Text]
[PDF]
|
|
|
|
|
|
|
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|
|
|
|
|
|
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[Full Text]
[PDF]
|
|
|
|
|
|
|
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[Full Text]
[PDF]
|
|
|
|
|
|
|
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105(21):
7594 - 7599.
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[Full Text]
[PDF]
|
|
|
|
|
|
|
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[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
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2075 - 2088.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
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[PDF]
|
|
|
|
|
|
|
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179(6):
1219 - 1230.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
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J. Neurosci.,
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27(34):
9201 - 9219.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
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Therapeutic effects of immunization with mutant superoxide dismutase in mice models of amyotrophic lateral sclerosis
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February 13, 2007;
104(7):
2495 - 2500.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
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Pyrrolidine Dithiocarbamate Inhibits Induction of Immunoproteasome and Decreases Survival in a Rat Model of Amyotrophic Lateral Sclerosis
Mol. Pharmacol.,
January 1, 2007;
71(1):
30 - 37.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
D. R. Beers, J. S. Henkel, Q. Xiao, W. Zhao, J. Wang, A. A. Yen, L. Siklos, S. R. McKercher, and S. H. Appel
Wild-type microglia extend survival in PU.1 knockout mice with familial amyotrophic lateral sclerosis
PNAS,
October 24, 2006;
103(43):
16021 - 16026.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
T. Kabuta, Y. Suzuki, and K. Wada
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J. Biol. Chem.,
October 13, 2006;
281(41):
30524 - 30533.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
S. Kalra, C. C. Hanstock, W. R. W. Martin, P. S. Allen, and W. S. Johnston
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August 1, 2006;
63(8):
1144 - 1148.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
S. Boillee, K. Yamanaka, C. S. Lobsiger, N. G. Copeland, N. A. Jenkins, G. Kassiotis, G. Kollias, and D. W. Cleveland
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June 2, 2006;
312(5778):
1389 - 1392.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
C. M. Schonhoff, M. Matsuoka, H. Tummala, M. A. Johnson, A. G. Estevéz, R. Wu, A.és Kamaid, K. C. Ricart, Y. Hashimoto, B. Gaston, et al.
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PNAS,
February 14, 2006;
103(7):
2404 - 2409.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
P. A. Jonsson, K. S. Graffmo, P. M. Andersen, T. Brannstrom, M. Lindberg, M. Oliveberg, and S. L. Marklund
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Brain,
February 1, 2006;
129(2):
451 - 464.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
F. E. Perrin, G. Boisset, M. Docquier, O. Schaad, P. Descombes, and A. C. Kato
No widespread induction of cell death genes occurs in pure motoneurons in an amyotrophic lateral sclerosis mouse model
Hum. Mol. Genet.,
November 1, 2005;
14(21):
3309 - 3320.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
J. Wang, G. Xu, H. Li, V. Gonzales, D. Fromholt, C. Karch, N. G. Copeland, N. A. Jenkins, and D. R. Borchelt
Somatodendritic accumulation of misfolded SOD1-L126Z in motor neurons mediates degeneration: {alpha}B-crystallin modulates aggregation
Hum. Mol. Genet.,
August 15, 2005;
14(16):
2335 - 2347.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
P J Shaw
Molecular and cellular pathways of neurodegeneration in motor neurone disease
J. Neurol. Neurosurg. Psychiatry,
August 1, 2005;
76(8):
1046 - 1057.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
J. Kirby, E. Halligan, M. J. Baptista, S. Allen, P. R. Heath, H. Holden, S. C. Barber, C. A. Loynes, C. A. Wood-Allum, J. Lunec, et al.
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Brain,
July 1, 2005;
128(7):
1686 - 1706.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
C. W. Strey, D. Spellman, A. Stieber, J. O. Gonatas, X. Wang, J. D. Lambris, and N. K. Gonatas
Dysregulation of Stathmin, a Microtubule-Destabilizing Protein, and Up-Regulation of Hsp25, Hsp27, and the Antioxidant Peroxiredoxin 6 in a Mouse Model of Familial Amyotrophic Lateral Sclerosis
Am. J. Pathol.,
November 1, 2004;
165(5):
1701 - 1718.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
C. Vanoni, S. Massari, M. Losa, P. Carrega, C. Perego, L. Conforti, and G. Pietrini
Increased internalisation and degradation of GLT-1 glial glutamate transporter in a cell model for familial amyotrophic lateral sclerosis (ALS)
J. Cell Sci.,
October 15, 2004;
117(22):
5417 - 5426.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
L. Dupuis, H. Oudart, F. Rene, J.-L. G. de Aguilar, and J.-P. Loeffler
Evidence for defective energy homeostasis in amyotrophic lateral sclerosis: Benefit of a high-energy diet in a transgenic mouse model
PNAS,
July 27, 2004;
101(30):
11159 - 11164.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
S. P. Koushika, A. M. Schaefer, R. Vincent, J. H. Willis, B. Bowerman, and M. L. Nonet
Mutations in Caenorhabditis elegans Cytoplasmic Dynein Components Reveal Specificity of Neuronal Retrograde Cargo
J. Neurosci.,
April 21, 2004;
24(16):
3907 - 3916.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
M. D. Nguyen, T. D'Aigle, G. Gowing, J.-P. Julien, and S. Rivest
Exacerbation of Motor Neuron Disease by Chronic Stimulation of Innate Immunity in a Mouse Model of Amyotrophic Lateral Sclerosis
J. Neurosci.,
February 11, 2004;
24(6):
1340 - 1349.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
P. A. Jonsson, K. Ernhill, P. M. Andersen, D. Bergemalm, T. Brannstrom, O. Gredal, P. Nilsson, and S. L. Marklund
Minute quantities of misfolded mutant superoxide dismutase-1 cause amyotrophic lateral sclerosis
Brain,
January 1, 2004;
127(1):
73 - 88.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
A. M. Clement, M. D. Nguyen, E. A. Roberts, M. L. Garcia, S. Boillee, M. Rule, A. P. McMahon, W. Doucette, D. Siwek, R. J. Ferrante, et al.
Wild-Type Nonneuronal Cells Extend Survival of SOD1 Mutant Motor Neurons in ALS Mice
Science,
October 3, 2003;
302(5642):
113 - 117.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
S. D. Rao, H. Z. Yin, and J. H. Weiss
Disruption of Glial Glutamate Transport by Reactive Oxygen Species Produced in Motor Neurons
J. Neurosci.,
April 1, 2003;
23(7):
2627 - 2633.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
R. J. Mockett, S. N. Radyuk, J. J. Benes, W. C. Orr, and R. S. Sohal
Phenotypic effects of familial amyotrophic lateral sclerosis mutant Sod alleles in transgenic Drosophila
PNAS,
January 7, 2003;
100(1):
301 - 306.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
D. Z. Ellis, J. Rabe, and K. J. Sweadner
Global Loss of Na,K-ATPase and Its Nitric Oxide-Mediated Regulation in a Transgenic Mouse Model of Amyotrophic Lateral Sclerosis
J. Neurosci.,
January 1, 2003;
23(1):
43 - 51.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
K. Puttaparthi, W. L. Gitomer, U. Krishnan, M. Son, B. Rajendran, and J. L. Elliott
Disease Progression in a Transgenic Model of Familial Amyotrophic Lateral Sclerosis Is Dependent on Both Neuronal and Non-Neuronal Zinc Binding Proteins
J. Neurosci.,
October 15, 2002;
22(20):
8790 - 8796.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
W. Sun, H. Funakoshi, and T. Nakamura
Overexpression of HGF Retards Disease Progression and Prolongs Life Span in a Transgenic Mouse Model of ALS
J. Neurosci.,
August 1, 2002;
22(15):
6537 - 6548.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
D. S. Howland, J. Liu, Y. She, B. Goad, N. J. Maragakis, B. Kim, J. Erickson, J. Kulik, L. DeVito, G. Psaltis, et al.
Focal loss of the glutamate transporter EAAT2 in a transgenic rat model of SOD1 mutant-mediated amyotrophic lateral sclerosis (ALS)
PNAS,
January 24, 2002;
(2002)
32539299.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
D. S. Howland, J. Liu, Y. She, B. Goad, N. J. Maragakis, B. Kim, J. Erickson, J. Kulik, L. DeVito, G. Psaltis, et al.
Focal loss of the glutamate transporter EAAT2 in a transgenic rat model of SOD1 mutant-mediated amyotrophic lateral sclerosis (ALS)
PNAS,
February 5, 2002;
99(3):
1604 - 1609.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
TOP
ABSTRACT
INTRODUCTION
