Neuroscience 145 (2007) 1233–1248
REVIEW
OXIDATIVE STRESS AND MITOCHONDRIAL DYSFUNCTION
IN NEURODEGENERATIVE DISEASES
E. TRUSHINAa AND C. T. MCMURRAYa,b,c*
a
Department of Molecular Pharmacology and Experimental Therapeu-
tics, Mayo Clinic, 200 First Street SW, Rochester, MN 55905, USA
b
Department of Biochemistry and Molecular Biology, Mayo Clinic, 200
First Street SW, Rochester, MN 55905, USA
c
Molecular Neuroscience Program, Mayo Clinic, 200 First Street SW,
Rochester, MN 55905, USA
Abstract—In recent years, it has become increasingly clear
that mitochondrial dysfunction and oxidative damage are
major contributors to neuronal loss. Free radicals, typically
generated from mitochondrial respiration, cause oxidative
damage of nucleic acids, lipids, carbohydrates and proteins.
Despite enormous amount of effort, however, the mechanism
by which oxidative damage causes neuronal death is not well
understood. Emerging data from a number of neurodegen-
erative diseases suggest that there may be common features
of toxicity that are related to oxidative damage. In this review,
while focusing on Huntington’s disease (HD), we discuss
similarities among HD, Friedreich ataxia and xeroderma pig-
mentosum, which provide insight into shared mechanisms of
neuronal death. © 2006 Published by Elsevier Ltd on behalf of
IBRO.
Key words: neurodegeneration, mitochondria, oxidative dam-
age, Huntington’s disease, Friedreich ataxia, xeroderma pig-
mentosum.
Contents
HD 1233
Features of HD pathophysiology 1234
Mhtt alters the functions of its interacting partners 1234
MT are dysfunctional in HD patients and in mouse models
for disease 1235
Possible mechanisms of MT dysfunction
and toxicity in HD 1238
Mutations in mtDNA and nuclear DNA 1238
Defects in MT functions 1238
Lipid/membrane damage 1239
*Corresponding author. Tel: ⫹1-507-284-1597; fax: ⫹1-507-284-9111.
E-mail address: mcmurray.cynthia@mayo.edu (C. T. McMurray).
Abbreviations: BDNF, brain-derived neurotrophic factor; CREB, cyclic
AMP response element-binding protein; cybrid, cytoplasmic hybrid;
FRDA, Friedreich ataxia; HAP1, huntingtin-associated protein 1; HD,
Huntington’s disease; HIP1, huntingtin interacting protein 1; Hippi,
huntingtin interacting protein-1 protein interactor; htt, huntingtin;
InsP3R1, inositol (1,4,5)-triphosphate receptor; mhtt, mutant hunting-
tin; MnSOD, manganese superoxide dismutase; MSN, medium spiny
neurons; MT, mitochondria; mtDNA, mitochondrial DNA; MT-PTP,
mitochondrial permeability transition pore; NER, nucleotide excision
repair; NPC, Niemann-Pick type C1; polyQ, polyglutamine; ROS, re-
active oxygen species; SDH, succinate dehydrogenase; SOD, super-
oxide dismutase; TCR, transcription-coupled repair; XP, xeroderma
pigmentosum; YAC, yeast artificial chromosome.
0306-4522/07$30.00⫹0.00 © 2006 Published by Elsevier Ltd on behalf of IBRO.
doi:10.1016/j.neuroscience.2006.10.056
1233
Mutations with direct effects on MT Function: FRDA 1240
Features of disease 1240
Abnormal regulation of antioxidants in MT in FRDA 1241
Mutations with direct effect on oxidative DNA damage 1242
DNA damage response as a common link 1242
Therapeutic implications of oxidative damage 1242
Acknowledgment 1243
References 1243
In humans, oxidative DNA damage is considered an im-
portant promoter of neurological diseases and aging
(Singh, 2006; Poulsen, 2005). In the cell, reactive oxygen
species (ROS) are generated primarily by mitochondria
(MT) (Reddy, 2006). To ensure efficient removal of MT-
generated ROS, the inner mitochondrial membrane incor-
porates a number of free radical scavengers such as vita-
min E, ascorbate, and glutathione. Additionally, there is an
enzymatic removal of free radicals by superoxide dis-
mutase (SODs). These lines of defense insure that forma-
tion of ROS as a by-product of respiration does not dam-
age the cell. The defense can be compromised by either
genetic mutations leading to decreased activity of the an-
tioxidants as in some familial cases of amyotrophic lateral
sclerosis (ALS) with mutation in CuZn SOD (Mariani et al.,
2005), or by increased radical production. In either case,
oxidative damage can impart harmful consequences to the
cell (Harman, 1956).
The mechanism by which oxidative damage causes
neuronal death is poorly understood. Therefore, an unre-
solved issue is whether mitochondrial defects are the pri-
mary cause of toxicity or a secondary response to the
damage. In this review, we consider characteristics of
three classes of neurodegenerative diseases with differ-
ent, but overlapping links to oxidative damage: Hunting-
ton’s disease (HD), Friedreich ataxia (FRDA), and xero-
derma pigmentosum (XP) (Fig. 1). XP arises directly from
inability to remove some forms of oxidative DNA damage,
FRDA arises from defects in MT and promiscuous Fenton
chemistry, and HD is representative of an “aggregation”
disorder where oxidative damage and MT defects are
thought to cause and/or contribute to toxicity. The goal of
the review is to gain insight into whether oxidative damage
is a common denominator for neurodegeneration.
HD
HD is a hereditary autosomal dominant disorder in which
the underlying mutation is an expanded CAG repeat in
exon 1 of the coding region of the HD gene (Fig. 2A). The
1234 E. Trushina and C. T. McMurray / Neuroscience 145 (2007) 1233–1248

Fig. 1. Oxidative DNA damage in neurodegenerative disorders can be caused directly and/or indirectly through altered mitochondrial function. Three
hereditary diseases mediate oxidative DNA damage and neuronal death through different pathways. Expression of mhtt in HD can directly increase
production of ROS and subsequent oxidative DNA damage through defects in MT. In FRDA, diminished levels of frataxin, a mitochondrial iron-binding
protein, directly affect MT function and ROS production. Lack of frataxin allows unbound iron to participate in Fenton chemistry with subsequent
generation of toxic hydroxyl radicals. In XP, neuronal death is caused by mutations in nucleotide excision repair proteins that interfere with the ability
to remove and repair oxidative DNA damage.

CAG triplet encodes the amino acid glutamine in the gene
product called huntingtin (htt) (The Huntington’s Disease
Collaborative Group, 1993). The mutant huntingtin (mhtt)
has elongated polyglutamine (polyQ) stretch in the N-ter-
minal region (⬎37 units) that affects its interaction with
htt-binding proteins and makes them susceptible to aggre-
gation (Harjes and Wanker, 2003). The onset of HD usu-
ally happens in the fourth and fifth decades of life. The
phenotype is characterized by loss of medium spiny neu-
rons (MSN) in the striatum, cognitive deterioration and
motor dysfunction (van Dellen et al., 2005) (Fig. 2B). The
CAG expansion process is dynamic, and growth of the
CAG tract can occur in the germ cells and in somatic cells
with age. The elongated polyQ tract generates mhtt with
increasingly aberrant properties (Lee and Kim, 2006;
Michalik and Van Broeckhoven, 2003). In patients, the
progressive toxicity of HD is manifested by a phenomenon
called anticipation where the onset of the disease inversely
correlates with the length of the polyQ tract (The Hunting-
ton’s Disease Collaborative Group, 1993).
While mutation analysis and transgenic animal models
for HD have unequivocally identified the expanded polyQ
tract as key for toxicity (Lee and Kim, 2006; Michalik and
Van Broeckhoven, 2003), the mechanism by which mhtt
progressively kills brain cells is poorly understood. PolyQ
regions self-associate to form polar zippers, the ␤-strands
capable to assemble into sheets or barrels using hydrogen
bonds and promote aggregation (Ross and Poirier, 2004;
Perutz, 1999). In HD-affected areas of the caudate and
cortex, immunochemical detection reveals that mhtt forms
high molecular weight complexes and inclusion bodies.
Thus, despite an incomplete understanding of toxic mech-
anism, it is generally accepted that in HD alteration and/or
sequestration of cellular targets are likely to be early caus-
ative events of the neuronal dysfunction and death (Lee
and Kim, 2006; Michalik and Van Broeckhoven, 2003;
Ross and Poirier, 2004).
Features of HD pathophysiology
Mhtt alters the functions of its interacting partners.
Proteins targeted by mhtt are presumably inactivated by it
or assume an alternate conformation (Harjes and Wanker,
2003; Li and Li, 2004) (Fig. 3). Indeed, mhtt modifies the
functions of many cytoplasmic and nuclear proteins often
found in aggregates (Harjes and Wanker, 2003; Li and Li,
2004). The N-terminal, truncated form of mhtt containing
polyQ tract can bind to and interfere with nuclear transcrip-
tion factors such as cyclic AMP response element-binding
protein (CREB) (Sugars et al., 2004), CREB binding pro-
tein (CBP) (Sugars et al., 2004; Rao and Finkbeiner, 2003;
Steffan et al., 2000), p53 (Steffan et al., 2000; Sawa, 2001;
Bae et al., 2005; Feng et al., 2006), co-repressor (Kegel et
al., 2002), transcriptional activator Sp1 and TAFII130 (Du-
nah et al., 2002; Li et al., 2002). Disruption of CREB in the
MT matrix in neurons decreases the expression of a sub-
set of mitochondrial genes, including the ND5 subunit of
complex I, and down-regulates complex I-dependent mito-
chondrial respiration. Thus, mhtt can sequester CREB or
any of its interacting partners by direct binding in either
 E. Trushina and C. T. McMurray / Neuroscience 145 (2007) 1233–1248 1235

Fig. 2. Expression of mhtt causes selective neuronal loss in striatum. (A) The HD gene has a polymorphic region consisting of the CAG repeats in
exon 1. In normal individuals, the number of CAG repeats is typically below 35. Growth of the CAG repeats in the coding region of HD gene produces
mutant protein with elongated polyQ region and aberrant functions. CAG expansion is progressive; the length of polyQ directly correlates with the
severity of HD and inversely with the onset of the disease. (B) Magnetic resonance image of normal (left) and HD brain (right). Expression of mhtt
causes neuronal loss in the striatum (caudate (C) and putamen (P)) and cortex as the disease progresses. Relative to the normal brain, substantial
neuronal loss is observed in the C and P, typical of an advanced stage (grade IV) of HD. Additionally, diseased brains are characterized by large
invaginations in the cortex and general brain atrophy. GP, globus pallidus: V, ventricle; hip, hippocampus.

nuclear or MT compartments (Shimohata et al., 2000;
Wyttenbach et al., 2001; Nucifora et al., 2001; Gines et al.,
2003).
Accumulating data suggest that a particularly important
effect of mhtt expression is disruption of microtubules
(Trushina et al., 2003) and inhibition of vesicular trafficking.
Htt normally interacts with trafficking motors and clathrin-
interacting proteins including Huntingtin interacting protein
1 (HIP1), Huntingtin interacting protein-related (HIP1R),
and Huntingtin-associated protein 1 (HAP1) (Harjes and
Wanker, 2003; Li et al., 1998; Engelender et al., 1997;
Tukamoto et al., 1997; Waelter et al., 2001). Consistent
with these protein interactions, htt appears to be involved
in both axonal trafficking and endocytosis, and inhibitory
effects of mhtt on axonal trafficking have been directly
demonstrated in squid axoplasm (Szebenyi et al., 2003), in
Drosophila (Gunawardena et al., 2003) and in mice
(Trushina et al., 2004) (Fig. 4). For example, htt together
with HAP1 enhances microtubule-based trafficking of an
important neuronal growth factor, brain-derived neurotro-
phic factor (BDNF), and expression of mhtt interferes with
that process (Gauthier et al., 2004). Mhtt binds with higher
affinity to HAP1 relative to htt, and interferes with BDNF
trafficking by competing with HAP1 for microtubule binding
(Gauthier et al., 2004). HAP1 can also serve as an adaptor
between htt and inositol (1,4,5)-triphosphate receptor
(InsP3R1) in lipid bilayers of MSN (Tang et al., 2003). In
the presence of mhtt, HAP1 sensitizes calcium release by
the InsP3R1 leading to the increase in the intracellular
calcium load, which, in turn, renders striatal neurons more
susceptible to toxicity (Tang et al., 2004).
Mhtt also has altered binding affinity to HIP1. In this
case, binding of mhtt is less strong relative to its normal
counterpart. The weaker interaction of mhtt with HIP1
shifts the equilibrium toward free HIP1, which can interact
with associated factors such as Hippi (huntingtin interact-
ing protein-1 protein interactor) (Tang et al., 2004; Majum-
der et al., 2006). Hippi forms a heterodimeric complex with
HIP1 and, under conditions of mhtt over-expression, the
HIP1/Hippi complex activates caspase-8 and initiates a
cascade of events leading to cell death. Steps include
cleavage of death agonist protein Bid, release of cyto-
chrome c and apoptosis inducing factor from MT, activa-
tion of caspase-9/caspase-6 and nuclear fragmentation
(Majumder et al., 2006). The fact that Hippi activates
caspase-6 is significant since proteolytic cleavage of mhtt
at the caspase-6 site appears to be a critical intermediate
of cell death (Graham et al., 2006).
MT are dysfunctional in HD patients and in mouse
models for disease. Perhaps, most relevant to oxidative
damage is the fact that mhtt directly impairs motility of MT.
Real-time imaging of primary neurons from animals ex-
pressing mhtt with 72 polyQ reveals that the movement of
elongated particles that by size and morphology resemble
MT (1.9 – 4.5 ␮m) is inhibited. The identity of the elongated
particles as MT has been confirmed by tetramethylrho-
damine, methyl ester staining (Fig. 4). Indeed, the motility
1236 E. Trushina and C. T. McMurray / Neuroscience 145 (2007) 1233–1248

Fig. 3. Cellular dysfunction caused by mhtt expression. Multiple protein–protein interactions and biological processes are altered in cells expressing
mhtt. Wild-type htt interacts with a large number of proteins that directly or indirectly regulate MT function and biogenesis. Mhtt has altered binding
affinity with interacting partners commonly affecting their functions. Diagram represents proteins and cellular functions that are affected by expression
of mhtt and could have an impact on MT dysfunction. Defective MT respond by over-production of ROS, and additional damage to MT and DNA would
be predicted. A vicious cycle eventually lead to neuronal damage and cell death.

of MT was strikingly impaired, and MT slowed down and
stopped frequently in primary striatal neurons from mice
expressing mhtt. Remarkably, MT in neurons from HD
animals travel distances 70% shorter than their normal
counterparts in the retrograde direction (Trushina et al.,
2004). It is not yet clear whether immobilization of MT
leads to their eventual dysfunction. However, MT that lose
their capacity to traverse the neurite heighten glutamate
excitotoxicity and alter calcium handling (Chang et al.,
2006), and inability to move could result in insufficient local
delivery of ATP.
In postmortem tissue from HD patients, severe defi-
ciencies in mitochondrial complex II and III and, some-
times, complex IV were found in the caudate (Mann et al.,
1990; Gu et al., 1996). Defects in complex II may be most
important for toxicity. Expression of both full-length and
N-terminal fragments of mhtt causes increased sensitivity
to complex II inhibitor 3-nitropropionic acid, while the ef-
fects of complex I inhibition do not induce cell death (Ruan
et al., 2004; Benchoua et al., 2006). Inhibition of MT com-
plex II appears to impart dysfunction by reducing the levels
of complex II/succinate dehydrogenase (SDH) subunits Ip
and Fp. As might be expected in this circumstance, the
SDH activity of the complex is diminished. Thus, in vitro,
treatment with 3-nitropropionic acid recapitulates MT de-
fects observed in human striatum and associated with
human disease. No major changes in the expression of
SDH subunits are found in the cerebral cortex and cere-
bellum, two regions less vulnerable to degeneration in HD
patients (Benchoua et al., 2006).
In transgenic animal models for HD, MT trafficking
defects are observed early in the embryo and appear
before other signs of toxicity (Trushina et al., 2004). How-
ever, MT dysfunction progresses with development and is
evident in older tissues by at least two measures: (1) the
diminished ATP production, and (2) the increased lactate
levels (a signature of MT dysfunction). Both have been
associated with HD in humans and in mouse models. A
decrease in succinate oxidation ranging from 39 to 59%
also occurs in the striatum of HD patients (Browne et al.,
1997; Tabrizi et al., 1999), and elevated lactate levels
correlate with CAG repeat length in HD occipital cortex and
striatum (Koroshetz et al., 1997; Jenkins et al., 1998).
These energetic defects have been observed even in
asymptomatic HD gene carriers (Jenkins et al., 1998).
Perhaps not surprising, HD-related MT dysfunction is also
observed in other tissues with higher ATP consumption
such as skeletal muscles. Myoblasts from presymptomatic
and symptomatic HD subjects are characterized by cellular
abnormalities in vitro such as mitochondrial depolarization,
cytochrome c release, and increased caspase-3, -8, and -9
activities. Although initially thought to exist exclusively in
the brain, mhtt-containing inclusion bodies are present in
differentiated myotubes (Ciammola et al., 2006). In HD
 E. Trushina and C. T. McMurray / Neuroscience 145 (2007) 1233–1248 1237

Fig. 4. Expression of mhtt inhibits trafficking of vesicles and MT in primary striatal neurons. (Top) Schematic diagram of axonal trafficking in neurons.
MT and vesicles are transported toward (retrograde) or away (anterograde) from the cells body. These trafficking functions influence secretion, Golgi
processing, MT biology, and energetic. (Bottom) Real time imaging of MT in axons in striatal neurons from mice expressing full-length mhtt. For
experiments with MT trafficking in living neurons, cells were treated for 5 min with tetramethylrhodamine dye (TMRM) (Invitrogen Corporation,
Carlsbad, CA, USA). The experiments were performed using confocal microscope LSM 510 (Carl Zeiss Inc., Oberkochen, Germany) with a
Plan-Apochromat 100⫻ (1.4 na) oil objective. Ar/Kr laser was set up to 568 nm for excitation; emission was set up to 585 nm and greater. TMRM was
washed away with fresh F-12K medium prior to imaging. Images were taken every 1 s at highest scan speed (0.9 s) for 10 min. A total of 600 images
was recorded per cell. Trafficking was analyzed using LSM 510 software that allowed animation of 600 images into a “movie.” Representative striatal
neuron (transmitted image t⫽0 s) and MT movement is shown on fluorescent images (t⫽0, 12, and 21 s). Reduced MT trafficking could cause
insufficient energy supply to the axonal growing cones and synaptic sites contributing to the neuronal dysfunction and altered synaptic transmission.
Aggregation observed later with disease progression can also significantly contribute to the defective MT trafficking.

pre-symptomatic and symptomatic patients, the effects in
muscles are accompanied by a decrease in complex I
activity, a decrease in phosphocreatine concentrations at
rest, and a reduction in ATP synthesis during exercise
(Lodi et al., 2000; Arenas et al., 1998; Saft et al., 2005).
The energetic defects observed in humans are repli-
cated in rodent models for disease. Similar to humans,
systemic administration of the mitochondrial complex II
inhibitor, 3-nitropropionic acid in rats, causes neurobehav-
ioral and pathological abnormalities consistent with HD
(Beal et al., 1991). In muscles of the R6/2 animal model of
HD, cytochrome c oxidase activity is reduced and abnor-
mal complex I-dependent respiration of MT is observed
(Gizatullina et al., 2006). Lactate is significantly increased
in the striatum from adult yeast artificial chromosome
(YAC) 72 mice relative to embryo (Trushina et al., 2004).
These in vitro data are consistent with those observed in
patients, and together implicate MT damage as relevant to
the disease.
If such a mechanism is pertinent in vivo, then an in-
crease in free radical production from defective MT might
initiate DNA damage in mhtt expressing animals. Indeed,
recent data indicate that this is the case. Over-expression
of free radical scavenger, Cu/Zn SOD1 reverses the oxi-
dative stress-induced proteasomal malfunction, aggrega-
tion, and cell death observed in HD (Goswami et al., 2006).
Increases in DNA lesions such as 8-hydroxy-2=-deox-
yguanosine (8-OHdG) and the extent of lipid peroxidation
have been useful markers of oxidative damage (Floyd et
al., 1990). In the R6/1 transgenic model for HD, progres-
sion of the neurological phenotype correlates with a pro-
gressive increase in striatal lipid peroxidation (Perez-
Severiano et al., 2000). Similarly, oxidative damage in
mitochondrial DNA (mtDNA) occurs in the parietal region
of human HD brain, but not frontal cortex or cerebellum
consistent with the regional specificity of neuronal vulner-
ability in HD (Polidori et al., 1999). Dysfunction in HD is
accompanied by a rise of p53 expression in brain cells
(Bae et al., 2005). P53 is a surveillance protein and tran-
scription factor that is “turned on” in response to DNA
damage. Thus, the up-regulation of p53 in animals and
cells strongly suggests that neuronal death may in part
involve an oxidative DNA damage response (Kruman et
al., 2004; Hazra et al., 2001; Bjelland and Seeberg, 2003).
Consistent with that notion, treatment of HD animals and
humans with anti-oxidants such as creatine (Ferrante et
al., 2000; Andreassen et al., 2001) and coenzyme Q (Fer-
rante et al., 2002; Schilling et al., 2001) has been benefi-
cial, although not curative, in preliminary clinical trials.
It seems clear that MT damage is an intermediate of
toxicity and promotes the release of destructive hydroxyl
radicals and loss of ATP production. Unfortunately, the
plethora of cellular effects due to mhtt expression leaves
unresolved the issue of whether MT damage is a primary
or secondary event in toxicity. Ironically, MT is both the
major source of ROS production, but also the major target
1238 E. Trushina and C. T. McMurray / Neuroscience 145 (2007) 1233–1248
of ROS damage. Therefore, oxidative damage to the MT
may destroy the special defense mechanisms that main-
tain functional integrity and protection against ROS. In MT,
superoxide is converted to the less reactive hydrogen per-
oxide by the mitochondrial matrix enzyme manganese su-
peroxide dismutase (MnSOD) (van Loon et al., 1986;
Guidot et al., 1993). Hydrogen peroxide levels are in turn
regulated by glutathione peroxidase (Takahashi et al.,
1987; Esposito et al., 2000) and catalase (Michiels et al.,
1994) activity, although the latter is not normally found in
the MT (Zhou and Kang, 2000). Any remaining hydrogen
peroxide is free to react with available iron to produce
hydroxyl radicals. The small amount of superoxide that is
not removed by MnSOD remains dangerous and can lead
to cell damage in two ways. One is to react with nitric oxide
to form reactive peroxynitrite (Huie and Padmaja, 1993), a
toxic species capable of inhibiting the respiratory complex
(Radi et al., 1994), as well as inactivating aconitase (Cas-
tro et al., 1994) and MnSOD by nitration of tyrosine resi-
dues (MacMillan-Crow et al., 1998). Alternatively, super-
oxide can directly inactivate certain Fe-S proteins (Kuo et
al., 1987; Gardner and Fridovich, 1991), particularly acon-
itase (Gardner et al., 1995), by stripping iron from the
prosthetic group. The released iron becomes available and
represents a secondary path by which superoxide can
influence rates of Fenton chemistry (Keyer and Imlay,
1996; McCormick et al., 1998). Mitochondrial impairment
and oxidative stress are detected even in asymptomatic
HD carriers early in HD progression (Saft et al., 2005), and
htt itself may be an iron-binding protein (Hilditch-Maguire
et al., 2000). Thus, it is difficult to determine what process
is responsible for initiation of the pathological cascade in
HD. Direct damage of DNA, aberrant MT functions, altered
calcium responses, and damage to lipids are all potentially
toxic. There is evidence to indicate that all contribute to
some extent to HD pathophysiology.
Possible mechanisms of MT dysfunction
and toxicity in HD
Mutations in mtDNA and nuclear DNA. MT are con-
trolled by two genomes: nuclear and mtDNA. Indeed, it has
been definitively shown that mutations in either nuclear or
mtDNA can contribute to MT dysfunction and neurodegen-
eration (Beal, 2005). The trinucleotide expansion, which
underlies HD, does not directly affect the expression of
proteins involved in oxidative phosphorylation, and, thus,
does not directly alter MT function. However, mtDNA is
predicted to be particularly vulnerable to ROS-induced
damage since it is located relatively close to the sites of
ROS generation. Several experimental studies have ad-
dressed the relationship between mtDNA damage and
dysfunction in HD, yet, to date, there is little evidence to
suggest that inherited mutations in mtDNA are a primary
cause of disease.
Scanning of mtDNA from members of the large Vene-
zuelan family with 43 restriction endonucleases failed to
reveal any differences in the mitochondrial genotype that
could account for the differences in age of onset between
the affected father and his progeny. In addition, analysis of
MT proteins from lymphoblast cell lines of these family
members revealed no detectable alterations. These data
suggest that the maternally inherited mitochondrial ge-
nome does not play a crucial role in determining age of
onset in HD (Irwin et al., 1989). Similar conclusions were
reached using cytoplasmic hybrid (cybrid) cells. Cybrid
cells lack their own MT. Thus, mtDNA from a human donor
can be reintroduced into these cells to test for functional
defects coded for by MT genome (King and Attardi, 1989).
However, cybrid cell lines containing mtDNA from HD pa-
tients do not differ from control cybrid lines in electron
transport chain, oxidative stress or calcium homeostasis
assays (Swerdlow et al., 1999). While mitochondrial dys-
function in HD does not appear to be due to inherited
mutations in mtDNA, somatic mutations in mtDNA may
play the important role. A common mtDNA4977 deletion is
markedly increased in the cortex of HD patients during
progression of disease (Horton et al., 1995) likely due to
oxidative damage mediated by faulty MT. Somatic mtDNA
mutations are particularly devastating to the striatal neu-
rons (Cantuti-Castelvetri et al., 2005), which accumulate
more mtDNA point mutations than any other brain cells.
These data suggest that oxidative damage and accumu-
lation of mtDNA mutations could contribute to toxicity dur-
ing progression of disease.
In fact, the importance of the DNA damage response in
toxicity is supported by the fact that loss of p53 in mhtt
expressing animals suppresses neurodegeneration (Bae
et al., 2005; Sawa, 2001; Feng et al., 2006). The response
to DNA damage must, at some level, be transmitted to MT
since p53 is a regulator of many MT genes. Interestingly,
the increase in nuclear DNA damage and up-regulation of
p53 activity in neurons is accompanied by an abnormal
flow of calcium ions across the MT membrane and in-
creased aggregate formation (Chang et al., 2006). Thus,
DNA damage and its detection by p53 appear to create a
feed-forward loop of toxicity that exacerbates the initial
effects of oxidative damage.
Defects in MT functions. Perhaps, the connection
between oxidative DNA damage and mhtt arises directly
from binding to MT. In cells expressing mhtt, the mitochon-
drial permeability transition pores (MT-PTP) respond to a
lower Ca2⫹ threshold (Panov et al., 2002; Choo et al.,
2004) (Fig. 5). In both humans and mouse models, the
observed dysfunction occurs in purified MT indicating that,
at least in part, the MT themselves are defective. Isolated
MT from lymphoblasts of HD patients, transgenic animals,
and immortalized striatal cells derived from HD knock-in
mice, consistently show more sensitivity in opening of the
MT-PTP in response to calcium. Moreover, HD cells are
also more sensitive to cyanide, a complex IV inhibitor, and
susceptible to apoptosis upon stress relative to normal
cells (Sawa et al., 1999; Panov et al., 2002). These defects
correlate directly with gene dosage. Lymphoblasts from
HD patients heterozygous or homozygous for the mutation
have the same age of disease onset, however, homozy-
gous patients display a more aggressive phenotype with
enlarged MT with calcium-sensitized membrane potential.
 E. Trushina and C. T. McMurray / Neuroscience 145 (2007) 1233–1248 1239

Fig. 5. Potential targets for ROS-generated oxidative damage in MT. MT-mediated neuronal dysfunction could be caused by disruption of the
mitochondrial membrane potential, sensitized MT-PTP, excitotoxin-induced Ca2⫹ influx, and diminished ATP production. In experimental models,
mitochondrial toxins (3-nitropropionic acid and malonate) impair respiratory enzyme activities and ATP generation and increase ROS. This is
consistent with impaired electron transport chain activities in HD. Superoxide radical (O2⫺) interacts with nitric oxide (NO) and produces peroxynitrite
(ONOO⫺). Both ROS and ONOO⫺ may then damage cellular macromolecules through nitrosylation, oxidation, and peroxidation, which can directly
contribute to neuronal injury. In addition, the release of cytochrome c from damaged/dysfunctional MT triggers the activation of apoptotic cascade and
release of initiator and executioner caspases (caspase-9 and -3), resulting in neuronal cell death. Calcium channels and ATP synthesis are also
affected.

Thus, early MT impairment at the basal level may affect the
severity of HD progression (Squitieri et al., 2006). Ultra-
structural examination of MT in cortical biopsies from the
patients with juvenile and adult-onset HD also display mor-
phological abnormalities (Browne and Beal, 2004). Due to
the sensitivity of the MT-PTP to calcium, it has been spec-
ulated that MT from HD patients might be less well
equipped to buffer calcium influxes into cells. Indeed, mhtt-
expressing cells are sensitive to glutamate receptor stim-
ulation, and import of extracellular calcium ions into the
cytoplasm. Glutamate receptors are abundantly expressed
in the MSN, the most affected cell type in HD. In striatal
MSN from YAC transgenic mouse model for HDg20 (Zeron
et al., 2002; Tang et al., 2005), glutamate released from
corticostriatal projection neurons stimulates NR1A/NR2B
N-methyl-D-aspartate receptor (NMDAR) and mGluR5 re-
ceptors. However, HD MSN are sensitized by InsP3R1 to
activation by InsP3, and once the mitochondrial Ca2⫹ stor-
age capacity is exceeded, the MT-PTP opens, leading to
the release of cytochrome c into the cytosol and activation
of caspases-9 and -3. In support of this model, glutamate
blockers reduce glutamate-induced apoptosis in the
YAC128 mouse model for HD (Leavitt et al., 2006) and
over-expression of full-length wild-type htt, which is known
to be neuroprotective (Cattaneo et al., 2001; Rigamonti et
al., 2000; Bezprozvanny and Hayden, 2004; Leavitt et al.,
2001), reduces excitotoxicity in response to glutamate
stimulation. Therefore, mhtt may promote excitotoxic death
by raising the calcium level to a concentration that exceeds
the buffering capacity of the MT (Beal, 1992).
There are, however, uncertainties with respect to both
calcium sensitivity and ATP production in mhtt-expressing
models (Fig. 5). Careful analyses of several mouse models
using sensitive electrodes indicate that the sensitivity to
calcium production is tissue-specific (Brustovetsky et al.,
2005). MT in striatum appear to be more sensitive to
calcium than are those in cortex, but there is no change
in calcium sensitivity with age (Brustovetsky et al., 2005).
In fact, the sensitivity to calcium load recovers with time in
transgenic animals expressing mhtt. It has also been re-
ported that expression of mhtt with 111 polyQ in clonal
striatal cells from knock-in HD mice causes significant
reduction in ATP production without directly affecting MT
complexes I–IV (Milakovic and Johnson, 2005). These
experiments appear to be in agreement with the data
obtained in the pre-symptomatic or grade I HD patients
(Guidetti et al., 2001), and findings in patient fibroblasts,
which demonstrate that ATP production declines in a linear
proportion with increasing polyQ length (Seong et al.,
2005). Thus, ATP production and the sensitivity to calcium
load may be separable. The discrepancy in the results
could be attributed either to the differences in the method-
ological approaches, cell or tissue types, and/or ages of
animals used. A careful and systematic analysis may be
necessary to resolve these issues.
Lipid/membrane damage. Incubation of MT with
GST-fused polyQ tracts of pathological length results in a
reduction in membrane potential and an increase in depo-
larization (Panov et al., 2003). No inhibition of any electron
transfer chain (ETC) complexes, mitochondrial ATP syn-
thase or adenine nucleotide translocase activities has
been observed in purified MT (Puranam et al., 2006). At
the same time, addition of antioxidants or cytochrome c
1240 E. Trushina and C. T. McMurray / Neuroscience 145 (2007) 1233–1248
prevents defects in isolated MT respiration caused by
polyQ proteins. Thus, toxic effect of polyQ proteins could
be in facilitating the significant increase in the amount of
ROS in isolated MT. The detailed mechanism of the inter-
action behind these processes remains to be elucidated.
However, one of the possible mechanisms could include
the increased ROS production facilitated by membrane-
associated oxidative stress (MAOS).
Brain MT have a higher concentration of lipids with
polyunsaturated acyls, which are more sensitive to free
radical oxidation than any other lipids (Bolanos et al.,
1997). Membrane lipid oxidation and oxidative modification
of membrane proteins in MT can cause diminished ATP
production with subsequent neuronal dysfunction (Matt-
son, 2004). Moreover, peroxidative damage to membrane
lipids is known to affect membrane viscosity and barrier
function (Mattson, 2004). Recent data suggest that both htt
and mhtt interact with lipid bilayers and MT membranes
(Kegel et al., 2000, 2005; Suopanki et al., 2006; Panov et
al., 2002). There is little evidence that lipid peroxidation of
MT by mhtt is a major feature of disease. However, the
potential effects could be severe. PolyQ expansion in mhtt
can alter its membrane localization (Kegel et al., 2005) and
can possibly interfere with the distribution and localization
of antioxidants present at the MT membrane. Mhtt might
also interfere with other membrane-associated events, e.g.
vesicle fission/fusion or endocytosis. Recently, association
of both htt and mhtt with brain lipids has been shown to
promote spontaneous aggregate formation in vivo (Suo-
panki et al., 2006). A similar phenomenon was recently
demonstrated for ␣-synuclein where binding to the plasma
membrane changed its structure, primed oligomerization
and accelerated fibrillogenesis (Cole et al., 2002; Jo et al.,
2004). Thus, the interaction of proteins with membrane
lipids could be of general importance in neurodegenerative
disorders.
Consistent with lipid abnormalities, a defect in choles-
terol metabolism has been linked to the MT dysfunction
and neurodegeneration in Niemann-Pick type C1 (NPC)
(Huang et al., 2006; Yu et al., 2005). Impaired cholesterol
transport in NPC neurons causes elevated cholesterol lev-
els in MT membranes leading to decreases in MT mem-
brane potential, ATP synthase activity, and markedly de-
creases the levels of ATP. Changes in the lipid composi-
tion of the MT membrane affect oxidative phosphorylation,
cell survival, by increasing the passive proton permeability
and membrane fluidity (Daum, 1985; Hoch, 1992). In con-
trast to other neurodegenerative disorders, in postmortem
tissues of HD patients and in fixed tissues from the R6/2
HD mice, the levels and synthesis of cholesterol are mark-
edly reduced (Valenza et al., 2005). Given, the similarities
among the degenerative diseases with respect to MT dys-
function, it is perhaps unusual that cholesterol content in
HD differs from that of the others, which is typically ele-
vated (Sjogren et al., 2006). It is possible that lipid metab-
olism is differentially affected in HD. Alternatively, reduc-
tion of cholesterol in HD may depend on the progression of
disease and/or the mouse model. Cholesterol reductions
have been measured only in postmortem human tissue or
animals with a very short lifespan. Therefore, it is possible
that loss of cholesterol is a feature of dying cells. Further
analysis will be required to distinguish among the
possibilities.
Whether MT defects are the primary cause of toxicity
or a secondary response in HD remains an unresolved
issue. The number of protein interactions altered by mhtt
association makes it difficult to cleanly separate effects of
altered function from the effects of oxidative damage (Fig.
3). However, neurodegeneration in several genetic dis-
eases arises more directly from MT dysfunction and oxi-
dative DNA damage, and may shed light on the relative
importance of oxidative damage in initiating neuronal
death.
MUTATIONS WITH DIRECT EFFECTS ON MT
FUNCTION: FRDA
Features of disease
FRDA is a hereditary recessive disorder directly associ-
ated with MT dysfunction and oxidative damage that arises
from inappropriate Fenton chemistry. The disease affects
roughly one in 50,000 people, and is characterized by
cardiac failure due to impaired ATP synthesis in cardiac
muscles (Lodi et al., 2006; Vorgerd et al., 2000). Atrophy of
sensory and cerebellar pathways causes ataxia, deep sen-
sory loss and loss of tendon reflexes. Corticospinal degen-
eration leads to muscular weakness, hypertrophic car-
diomyopathy may contribute to disability and cause pre-
mature death. The first symptoms usually appear in
childhood, but age of onset may vary from infancy to
adulthood. Individuals suffering from FRDA have life
expectancy of 38 years on average (Gatchel and
Zoghbi, 2005).
The FRDA gene encodes a protein of 210 amino acids
called frataxin (Campuzano et al., 1996). In humans,
frataxin mRNA is most abundant in the heart, spinal cord
and developing brain. Increasing evidence suggests that
this protein directs the intra-mitochondrial synthesis of
iron-sulfur (Fe/S) clusters (Lill and Muhlenhoff, 2005; Rötig
et al., 1997; Puccio et al., 2001; Muhlenhoff et al., 2002)
(Fig. 5). MT generate readily available energy equivalents
converting nutrients into ATP by oxidative phosphorylation.
This process is dependent on Fe/S clusters, which are
essential parts of MT enzymes including aconitase and
complexes I, II and III (Lill and Muhlenhoff, 2005). Although
the exact function of frataxin is only emerging, it is thought
to be a mitochondrial iron-binding protein that prevents this
metal from participating in Fenton chemistry and inappro-
priate generation of hydroxyl radicals (Fig. 6). Since FRDA
is caused by frataxin deficiency, it follows that loss of
frataxin is likely to enhance these events.
FRDA deficiency begins at the DNA level. Frataxin
contains a GAA triplet repeat expansion in the first intron of
the gene (Wilson, 2003). The disease is dominant reces-
sive, and most FRDA patients are homozygous for the
GAA expansion. Repeats in normal chromosomes contain
up to 40 triplets, while the disease allele can contain from
90 to thousands of GAA repeats (Gacy et al., 1998). In
 E. Trushina and C. T. McMurray / Neuroscience 145 (2007) 1233–1248
Fig. 6. Proposed consequences of frataxin deficiency in FRDA. A reduction in the levels of frataxin is predicted to affect mitochondrial iron metabolism,
with a decrease in the biosyntheses of heme and iron-sulfur clusters and a simultaneous increase in the labile iron pool. Labile Fe2⫹ no longer
detoxified by frataxin participates in Fenton chemistry leading to oxidative damage and progressive accumulation of ferric iron oxides.
about 2% of FRDA cases, however, one allele is expanded
and the other contains point mutations that alter the
frataxin coding sequence (Cossee et al., 1999). In either
case, affected individuals express little to no normal
frataxin, which is essential for survival. Like HD, FRDA
shows the anticipation phenotype (Montermini et al.,
1997a,b), and triplet expansion in the disease alleles both
arise from DNA secondary structure intermediates. In HD,
the CAG repeats form an aberrant hairpin structure, which
lies in the coding sequence of the gene, and results in the
HD protein with aberrant functions. In FRDA, the long GAA
tract resides in an intron, and does not code for a protein.
Rather, the GAA disease allele adopts a triple helical struc-
ture (Gacy et al., 1998; Ohshima et al., 1998), which acts
as a roadblock to transcription, translation, or splicing of
the mRNA, and creates a functional knockout of the protein
(Sakamoto et al., 2001).
As in HD, ROS-mediated damage and MT dysfunction
are associated with FRDA. MT in these patients progres-
sively lose respiratory competence, and cells become hy-
persensitive to hydrogen peroxide (Wong et al., 1999).
Consistent with this feature, FRDA patients have in-
creased plasma levels of lipid peroxidation (Emond et al.,
2000), oxidative DNA damage products, and hydroxyl rad-
ical production (Schulz et al., 2000) (Fig. 5). The increases
in oxidation products in FRDA appear to be related to im-
paired efflux of iron out of MT (Radisky et al., 1999). MT in
FRDA patients accumulates iron more then 10-fold in ex-
cess of wild type. Loss of respiratory competence requires
the presence of iron in the culture medium, and occurs
more rapidly as the iron concentration is increased. These
data suggest that permanent MT damage is the conse-
quence of iron toxicity and the elevation of oxidative dam-
age is due to inappropriate Fenton chemistry.
Neither Yfh1, the yeast homolog of frataxin, nor human
frataxin is regulated by iron levels, and their mRNAs do not
contain an iron responsive element. However, human
frataxin (Cavadini et al., 2002) and Yfh1p (Adamec et al.,
2000) are iron-binding proteins. Yfh1 appears to form a
high molecular weight complex in the presence of ferrous
ions (Adamec et al., 2000; Park et al., 2003). Thus, Yfh1p
and frataxin may protect iron in MT from contacts with free
radicals, and act as a mitochondrial iron chaperone (Bul-
teau et al., 2004). In the absence of frataxin, impaired
biosyntheses and transport processes may lead to iron
accumulation in a toxic, redox-active form (Fig. 6). Consis-
tent with this possibility, disruption of the frataxin homo-
logue in yeast increases oxidative stress (Gakh et al.,
2006; Wong et al., 1999; Piemonte et al., 2001; Chantrel-
Groussard et al., 2001) and sensitivity against oxidants, as
1241
well as promotes oxidative damage to both nuclear and
mtDNA (Karthikeyan et al., 2002, 2003). Similar to HD,
ATP synthesis in skeletal muscles from FRDA patients is
reduced after exercise, as measured by magnetic reso-
nance spectroscopy (Lodi et al., 1999), and ATP synthesis
inversely correlates with the length of the GAA tract.
Abnormal regulation of antioxidants in MT in FRDA
As for MT dysfunction, the same Fe-S defects found in
tissues from FRDA patients are present in yeast with def-
icits in respiratory complexes I, II and III, and aconitase
(Rötig et al., 1997) (Fig. 5). Although oxidative damage is
up regulated in FRDA, there is no induction of the detoxi-
fying enzyme MnSOD. This is surprising since MnSOD is
induced by a variety of factors that generate oxidative
stress, including proinflammatory cytokines such as tumor
necrosis factor (TNF) (Visner et al., 1990; Ono et al.,
1992). These data suggest that part of the toxic mecha-
nism in FRDA involves disruption of a regulatory pathway
for MnSOD. Under normal conditions, hydrogen peroxide
generation appears to be the rate-limiting factor in Fenton
chemistry, and prevents stripping of Fe-S proteins. Thus,
MnSOD activity is inversely correlated with hydroxyl radi-
cal production (Borrello et al., 1996; Manna et al., 1998;
Shi et al., 1999). Since free, soluble iron and iron bound in
Fe-S clusters account for the majority of MT iron, loss of
frataxin in FRDA cells is expected to increase the level
of free iron as is observed (Tan et al., 2001). A number of
recent studies indicate that frataxin has a role in heme
synthesis, not just iron–sulfur cluster synthesis/repair (Al-
Karadaghi et al., 2006). Frataxin appears to be a bifunc-
tional protein with the ability to bind ferrous iron and either
transfer it to other iron-binding proteins or detoxify it by
converting it to a stable ferrihydrite mineral. The latter is
inactive in ROS-generating reactions.
There are parallels between HD and FRDA. Mhtt is
also reported to be an iron binding protein, and iron de-
posits have been detected in the brain of HD patients
(Hilditch-Maguire et al., 2000). MnSOD induction in FRDA
is controlled by AP-1 and ATF-1/CREB-1 activation (Bor-
rello et al., 1992). CREB itself is a target of mhtt binding,
and if inactivated by mhtt, then MnSOD pathways may be
affected. The subsequent damage to the respiratory com-
plexes could lead to increased leakage of the superoxide
radicals that would account for the inactivation of Fe-S
proteins and DNA damage. Thus, MT dysfunction and
overlap in some properties of HD and FRDA have raised
the possibility that oxidative DNA damage might be a
common feature in neurodegenerative diseases and may
1242 E. Trushina and C. T. McMurray / Neuroscience 145 (2007) 1233–1248
directly contribute to neuronal toxicity. For FRDA, this hy-
pothesis is consistent with the fact that over-expression of
frataxin reduces intracellular accumulation of ROS, and
prevents menadione-induced malignant transformation of
fibroblasts (Shoichet et al., 2002). The loss of frataxin
function in FRDA also elevates ROS in cells, and mito-
chondrial dysfunction and a rise in oxidation is associated
with HD.
MUTATIONS WITH DIRECT EFFECT ON
OXIDATIVE DNA DAMAGE
A rise in oxidative DNA bases is observed in both HD and
FRDA. However, in both of these disorders a range of
cellular processes is affected. In the former, mhtt can
interact and inactivate key proteins associated with cyto-
plasmic and nuclear events, while in the latter iron metab-
olism appears to be altered. Thus, effects of faulty protein
function cannot be cleanly separated from oxidative DNA
damage, which, on its own, may be sufficient to elicit cell
death. The extent to which oxidative DNA damage contrib-
utes to cell death of neurons is not known. However,
lessons can be learned from a group of disorders referred
to as XP. XP arises from mutations in nucleotide excision
repair (NER) proteins, which are essential for removal of
some forms of oxidative DNA damage. NER is carried out
by a number of proteins that operate in discrete steps:
lesion recognition, single strand excision, lesion removal,
and, with the help of polymerase, gap-filling synthesis
(McMurray, 2005; Cleaver, 2005; Dip et al., 2004). NER is
characterized by two types of repair pathways: removal of
oxidative DNA damage found globally in the genome
(GGR), and oxidative DNA damage found in transcribed
genes (transcription coupled repair (TCR)) (Cleaver, 2005;
Dip et al., 2004). GGR and TCR share most steps in the
repair pathway, but differ in their recognition step. GGR is
initiated by recognition by XPC/HR23B or XPE/DDB2. In
contrast, TCR is initiated by two factors that are defective
in a form of XP called Cockayne’s syndrome (CS). CSA
and CSB signal TCR by recognizing a stalled RNA poly-
merase itself.
The NER pathway is associated with at least seven
complementary groups, XP (A–G), which represent muta-
tions in specific NER proteins. The incidence of neurolog-
ical abnormalities varies considerably. Neurodegeneration
is prominent in many XP patients but extremely rare in
patients with XPC or XPE. Since XPC and XPE are in-
volved in the initiation of GGR, these data indicate that
defects in TCR rather than GGR are connected to the
neurodegeneration phenotype. These data point to the
importance of keeping expressed genes lesion free. Oxi-
dative damage within expressed genes is more likely to be
associated with neuronal toxicity, not unexpectedly, given
the fact that neurons cannot divide and may accumulate
unrepaired DNA damage over time. In XP patients, faulty
NER and unrepaired oxidative DNA damage, on its own,
can result in death of neurons in the absence of additional
insults from defective proteins. The spectrum of ROS-
mediated lesions repaired by NER includes intrastrand
cross-links, bulky chemical lesions, cyclodeoxyadenosine,
thymine glycol, cyclopurines, and infrequently, 8-oxo-G
(Bjelland and Seeberg, 2003). Other DNA repair pathways
can remove oxidative damage to DNA due to ROS, notably
base excision repair. While some of these lesions can also
be removed by NER, in vivo, the major defect in neurons of
XP patients may be the inability to repair cyclopurines
(Hanawalt et al., 2003; Friedberg, 2003; Bootsma et al.,
2005). In any case, it appears that loss of repair of oxida-
tive DNA lesions alone can result in severe neurodegen-
erative disease with early onset in the absence of addi-
tional protein defects. Thus, the degree to which oxidative
DNA damage accumulates may be linked to the onset of
disease.
DNA DAMAGE RESPONSE AS A COMMON
LINK
The consequences of oxidative damage among neurode-
generative diseases are complex, and dysfunction can
occur at many levels. However, a common denominator is
likely to be an oxidative DNA damage response, which
appears to be sufficient by itself to promote neurodegen-
eration. In the case of HD, mhtt can rescue the develop-
mental defects of htt deletion (White et al., 1997). Thus,
mhtt must retain some of the functions of the normal pro-
tein. Moreover, HD displays later onset than does XP.
Thus, failure to remove oxidative damage in XP may be
more toxic than the effects of a faulty HD protein in other-
wise repair-competent neurons. Since repair of oxidative
lesions by either BER or NER occurs through a single
strand break intermediate, oxidative stress may cause tox-
icity through the accumulation of unrepaired single strand
breaks (El-Khamisy and Caldecott, 2006) and the possible
generation of double strand breaks. These data, as a
whole, raise the testable hypothesis that DNA damage,
when unchecked in neurons, is a common factor that
promotes toxicity.
THERAPEUTIC IMPLICATIONS OF OXIDATIVE
DAMAGE
Based on the evidence, it might be predicted that anti-
oxidants and/or inhibitors of excitotoxic damage might be
therapeutic. However, to date, the majority of HD preclin-
ical mouse trials designed to test the effect of different
neuroprotective agents demonstrated limited success
(Handley et al., 2006). Processes of free radical damage,
excitotoxicity and membrane integrity have been consid-
ered. Beneficial agents include creatine (Ferrante et al.,
2000; Andreassen et al., 2001), coenzyme Q (Ferrante et
al., 2002; Schilling et al., 2001), remacemide (Ferrante et
al., 2002; Schilling et al., 2001), riluzole (Schiefer et al.,
2002), a-lipoic acid (Andreassen et al., 2001) and essential
fatty acids (Clifford et al., 2002). Creatine treatment, for
example, is neuroprotective in R6/2 mice (Ferrante et al.,
2000). Similarly, administration of creatine to HD patients
for 2 years (10 g/day) prevented weight loss and, in some
patients, caused an improvement in the scores for neuro-
logical testing (Tabrizi et al., 2005). In an additional study,
 E. Trushina and C. T. McMurray / Neuroscience 145 (2007) 1233–1248
administration of creatine (8 g/day or 16 weeks) markedly
reduced serum levels of 8-hydroxy-2=-deoxyguanosine, an
indicator of oxidative injury to DNA (Hersch et al., 2006).
Small molecules that inhibit oxidative damage, such as
cystamine and cysteamine, have improved survival of
mhtt-expressing cells (Mao et al., 2006) and animal mod-
els for HD (Borrell-Pages et al., 2006) by preventing MT
depolarization and increasing BDNF levels in the brain. For
FRDA, Bayer 11–7082, a compound known to regulate
activity of the mitogen-activated protein kinases, JNK-1
and p38 (Read et al., 1997), is capable of inhibiting path-
ways of iron-mediated MnSOD induction. The existence of
a mechanism linking MnSOD regulation remains to be
proven but represents a logical link by which cellular ROS
defenses respond to changes in MT iron states. Although
none are curative, recent data continue to support benefi-
cial effects of antioxidants. As judged by dominant hered-
itary disorders, the data indicate that ROS are major con-
tributors to neuronal toxicity, and there is continued hope
that blocking ROS might be used to offset disease.
Acknowledgment—This work was supported by the Mayo Foun-
dation, the National Institutes of Health grants NS40738 (C.T.M.),
and GM 066359 (C.T.M.). We thank Kurt Johnson and Grazia
Isaya for reading the manuscript, and Grazia Isaya for provision of
Fig. 6.
REFERENCES
Adamec J, Rusnak F, Owen WG, Naylor S, Benson LM, Gacy AM,
Isaya G (2000) Iron-dependent self-assembly of recombinant
yeast frataxin: implications for Friedreich ataxia. Am J Hum Genet
67:549 –562.
Al-Karadaghi S, Franco R, Hansson M, Shelnutt JA, Isaya G, Ferreira
GC (2006) Chelatases: distort to select? Trends Biochem Sci
31:135–142.
Andreassen OA, Dedeoglu A, Ferrante RJ, Jenkins BG, Ferrante KL,
Thomas M, Friedlich A, Browne SE, Schilling G, Borchelt DR,
Hersch SM, Ross CA, Beal MF (2001) Creatine increase survival
and delays motor symptoms in a transgenic animal model of
Huntington’s disease. Neurobiol Dis 8:479 – 491.
Arenas J, Campos Y, Ribacoba R, Martin MA, Rubio JC, Ablanedo P,
Cabello A (1998) Complex I defect in muscle from patients with
Huntington’s disease. Ann Neurol 43:397– 400.
Bae BI, Xu H, Igarashi S, Fujimuro M, Agrawal N, Taya Y, Hayward
SD, Moran TH, Montell C, Ross CA, Snyder SH, Sawa A (2005)
p53 Mediates cellular dysfunction and behavioral abnormalities in
Huntington’s disease. Neuron 47:29 – 41.
Beal MF (1992) Does impairment of energy metabolism result in
excitotoxic neuronal death in neurodegenerative illnesses? Ann
Neurol 31:119 –130.
Beal MF (2005) Mitochondria take center stage in aging and neuro-
degeneration. Ann Neurol 58:495–505.
Beal MF, Ferrante RJ, Swartz KJ, Kowall NW (1991) Chronic quino-
linic acid lesions in rats closely resemble Huntington’s disease.
J Neurosci 11:1649 –1659.
Benchoua A, Trioulier Y, Zala D, Gaillard MC, Lefort N, Dufour N,
Saudou F, Elalouf JM, Hirsch E, Hantraye P, Deglon N, Brouillet E
(2006) Involvement of mitochondrial complex II defects in neuronal
death produced by N-terminus fragment of mutated huntingtin. Mol
Biol Cell 17:1652–1663.
Bezprozvanny I, Hayden MR (2004) Deranged neuronal calcium sig-
naling and Huntington disease. Biochem Biophys Res Commun
322:1310 –1317.
1243
Bjelland S, Seeberg E (2003) Mutagenicity, toxicity and repair of DNA
base damage induced by oxidation. Mutat Res 531:37– 80.
Bolanos JP, Almeida A, Fernandez E, Medina JM, Land JM, Clark JB,
Heales SJ (1997) Potential mechanisms for nitric oxide-mediated
impairment of brain mitochondrial energy metabolism. Biochem
Soc Trans 25:944 –949.
Bootsma D, Kraemer KH, Cleaver JE, Hoeijmakers JHJ (2005) Nucle-
otide excision repair syndromes: xeroderma pigmentosum, cock-
ayne syndrome, and trichothiodystrophy. In: The metabolic and
molecular bases of inherited disease, Vol. 3 (Scriver C, Beaudet
AL, Sly WS, eds), pp 137–150. McGraw-Hill Companies.
Borrell-Pages M, Canals JM, Cordelieres FP, Parker JA, Pineda JR,
Grange G, Bryson EA, Guillermier M, Hirsch E, Hantraye P,
Cheetham ME, Neri C, Alberch J, Brouillet E, Saudou F, Humbert
S (2006) Cystamine and cysteamine increase brain levels of BDNF
in Huntington disease via HSJ1b and transglutaminase. J Clin
Invest 116:1410 –1424.
Borrello S, De Leo ME, Landriscina M, Palazzotti B, Galeotti T (1996)
Diethyldithiocarbamate treatment up regulates manganese super-
oxide dismutase gene expression in rat liver. Biochem Biophys
Res Commun 220:546 –552.
Borrello S, De Leo ME, Wohlrab H, Galeotti T (1992) Manganese
deficiency and transcriptional regulation of mitochondrial superox-
ide dismutase in hepatomas. FEBS Lett 310:249 –254.
Browne SE, Beal MF (2004) The energetics of Huntington’s disease.
Neurochem Res 29:531–546.
Browne SE, Bowling AC, MacGarvey U, Baik MJ, Berger SC, Muqit
MM, Bird ED, Beal MF (1997) Oxidative damage and metabolic
dysfunction in Huntington’s disease: selective vulnerability of the
basal ganglia. Ann Neurol 41:646 – 653.
Brustovetsky N, LaFrance R, Purl KJ, Brustovetsky T, Keene CD, Low
WC, Dubinsky JM (2005) Age-dependent changes in the calcium
sensitivity of striatal mitochondria in mouse models of Huntington’s
disease. J Neurochem 93:1361–1370.
Bulteau AL, O’Neill HA, Kennedy MC, Ikeda-Saito M, Isaya G, Szweda
N (2004) Frataxin acts as an iron chaperone protein to modulate
mitochondrial aconitase activity. Science 305:242–245.
Campuzano V, Montermini L, Molto MD, Pianese L, Cossee M, Cav-
alcanti F, Monros E, Rodius F, Duclos F, Monticelli A, Zara F,
Canizares J, Koutnikova H, Bidichandani SI, Gellera C, Brice A,
Trouillas P, De Michele G, Filla A, De Frutos R, Palau F, Patel PI,
Di Donato S, Mandel JL, Cocozza S, Koenig M, Pandolfo M (1996)
Friedreich’s ataxia: autosomal recessive disease caused by an
intronic GAA triplet repeat expansion. Science 271:1423–1427.
Cantuti-Castelvetri I, Lin MT, Zheng K, Keller-McGandy CE, Betensky
RA, Johns DR, Beal MF, Standaert DG, Simon DK (2005) Somatic
mitochondrial DNA mutations in single neurons and glia. Neurobiol
Aging 26:1343–1355.
Castro L, Rodriguez M, Radi R (1994) Aconitase is readily inactivated
by peroxynitrite, but not by its precursor, nitric oxide. J Biol Chem
269:29409 –29415.
Cattaneo E, Rigamonti D, Goffredo D, Zuccato C, Squitieri F, Sipione
S (2001) Loss of normal huntingtin function: new developments in
Huntington’s disease research. Trends Neurosci 24:182–188.
Cavadini P, O’Neill HA, Benada O, Isaya G (2002) Assembly and
iron-binding properties of human frataxin, the protein deficient in
Friedreich ataxia. Hum Mol Genet 11:217–227.
Chang DT, Rintoul GL, Pandipati S, Reynolds IJ (2006) Mutant hun-
tingtin aggregates impair mitochondrial movement and trafficking
in cortical neurons. Neurobiol Dis 22:388 – 400.
Chantrel-Groussard K, Geromel V, Puccio H, Koenig M, Munnich A,
Rötig A, Rustin P (2001) Disabled early recruitment of antioxidant
defenses in Friedreich’s ataxia. Hum Mol Genet 10:2061–2067.
Choo YS, Johnson GV, MacDonald M, Detloff PJ, Lesort M (2004)
Mutant huntingtin directly increases susceptibility of mitochondria
to the calcium-induced permeability transition and cytochrome c
release. Hum Mol Genet 13:1407–1420.
1244 E. Trushina and C. T. McMurray / Neuroscience 145 (2007) 1233–1248
Ciammola A, Sassone J, Alberti L, Meola G, Mancinelli E, Russo MA,
Squitieri F, Silani V (2006) Increased apoptosis, huntingtin inclu-
sions and altered differentiation in muscle cell cultures from Hun-
tington’s disease subjects. Cell Death Differ 13:2068 –2078.
Cleaver JE (2005) Cancer in xeroderma pigmentosum and related
disorders of DNA repair. Nat Rev Cancer 5:564 –573.
Clifford JJ, Drago J, Natoli AL, Wong JY, Kinsella A, Waddington JL,
Vaddadi KS (2002) Essential fatty acids given from conception
prevent topographies of motor deficit in a transgenic model of
Huntington’s disease. Neuroscience 109:81– 88.
Cole NB, Murphy DD, Grider T, Rueter S, Brasaemle D, Nussbaum RL
(2002) Lipid droplet binding and oligomerization properties of the
Parkinson’s disease protein alpha-synuclein. J Biol Chem 277:
6344 – 6352.
Cossee M, Durr A, Schmitt M, Dahl N, Trouillas P, Allinson P, Kostr-
zewa M, Nivelon-Chevallier A, Gustavson KH, Kohlschutter A,
Muller U, Mandel JL, Brice A, Koenig M, Cavalcanti F, Tammaro A,
De Michele G, Filla A, Cocozza S, Labuda M, Montermini L, Poirier
J, Pandolfo M (1999) Friedreich’s ataxia: point mutations and
clinical presentation of compound heterozygotes. Ann Neurol 45:
200 –206.
Daum G (1985) Lipids of mitochondria. Biochim Biophys Acta
822:1– 42.
Dip R, Camenisch U, Naegeli H (2004) Mechanisms of DNA damage
recognition and strand discrimination in human nucleotide excision
repair. DNA Repair (Amst) 3:1409 –1423.
Dunah AW, Jeong H, Griffin A, Kim YM, Standaert DG, Hersch SM,
Mouradian MM, Young AB, Tanese N, Krainc D (2002) Sp1 and
TAFII130 transcriptional activity disrupted in early Huntington’s
disease. Science 296:2238 –2243.
El-Khamisy SF, Caldecott KW (2006) TDP1-dependent DNA single-
strand break repair and neurodegeneration. Mutagenesis 21(4):
219 –224.
Emond M, Lepage G, Vanasse M, Pandolfo M (2000) Increased levels
of plasma malondialdehyde in Friedreich ataxia. Neurology 55:
1752–1753.
Engelender S, Sharp AH, Colomer V, Tokito MK, Lanahan A, Worley
P, Holzbaur EL, Ross CA (1997) Huntingtin-associated protein 1
(HAP1) interacts with the p150Glued subunit of dynactin. Hum Mol
Genet 6:2205–2212.
Esposito LA, Kokoszka JE, Waymire KG, Cottrell B, MacGregor GR,
Wallace DC (2000) Mitochondrial oxidative stress in mice lacking
the glutathione peroxidase-1 gene. Free Radic Biol Med 28:754 –
766.
Feng Z, Jin S, Zupnick A, Hoh J, de Stanchina E, Lowe S, Prives C,
Levine AJ (2006) p53 Tumor suppressor protein regulates the
levels of huntingtin gene expression. Oncogene 25:1–7.
Ferrante RJ, Andreassen OA, Dedeoglu A, Ferrante KL, Jenkins BG,
Hersch SM, Beal MF (2002) Therapeutic effects of coenzyme Q10
and remacemide in transgenic mouse models of Huntington’s dis-
ease. J Neurosci 22:1592–1599.
Ferrante RJ, Andreassen OA, Jenkins BG, Dedeoglu A, Kuemmerle S,
Kubilus JK, Kaddurah-Daouk R, Hersch SM, Beal MF (2000) Neu-
roprotective effects of creatine in a transgenic mouse model of
Huntington’s disease. J Neurosci 20:4389 – 4397.
Floyd RA, West MS, Eneff KL, Schneider JE, Wong PK, Tingey DT,
Hogsett WE (1990) Conditions influencing yield and analysis of
8-hydroxy-2=-deoxyguanosine in oxidatively damaged DNA. Anal
Biochem 188:155–158.
Friedberg EC (2003) DNA damage and repair. Nature 421:436 – 440.
Gacy AM, Goellner GM, Spiro C, Chen X, Gupta G, Bradbury EM,
Dyer RB, Mikesell MJ, Yao JZ, Johnson AJ, Richter A, Melancon
SB, McMurray CT (1998) GAA instability in Friedreich’s ataxia
shares a common, DNA-directed and intraallelic mechanism with
other trinucleotide diseases. Mol Cell 1:583–593.
Gakh O, Park S, Liu G, Macomber L, Imlay JA, Ferreira GC, Isaya G
(2006) Mitochondrial iron detoxification is a primary function of
frataxin that limits oxidative damage and preserves cell longevity.
Hum Mol Genet 15:467– 479.
Gardner PR, Fridovich I (1991) Superoxide sensitivity of the Esche-
richia coli 6-phosphogluconate dehydratase. J Biol Chem 266:
1478 –1483.
Gardner PR, Raineri I, Epstein LB, White CW (1995) Superoxide
radical and iron modulate aconitase activity in mammalian cells.
J Biol Chem 270:13399 –13405.
Gatchel JR, Zoghbi HY (2005) Diseases of unstable repeat expansion:
mechanisms and common principles. Nat Rev Genet 6:743–
755.
Gauthier LR, Charrin BC, Borrell-Pages M, Dompierre JP, Rangone H,
Cordelieres FP, De Mey J, MacDonald ME, Lessmann V, Humbert
S, Saudou F (2004) Huntingtin controls neurotrophic support and
survival of neurons by enhancing BDNF vesicular transport along
microtubules. Cell 118:127–138.
Gines S, Seong IS, Fossale E, Ivanova E, Trettel F, Gusella JF,
Wheeler VC, Persichetti F, MacDonald ME (2003) Specific pro-
gressive cAMP reduction implicates energy deficit in presymptom-
atic Huntington’s disease knock-in mice. Hum Mol Genet 12:
497–508.
Gizatullina ZZ, Lindenberg KS, Harjes P, Chen Y, Kosinski CM, Land-
wehrmeyer BG, Ludolph AC, Striggow F, Zierz S, Gellerich FN
(2006) Low stability of Huntington muscle mitochondria against
Ca2⫹ in R6/2 mice. Ann Neurol 59:407– 411.
Goswami A, Dikshit P, Mishra A, Mulherkar S, Nukina N, Jana NR
(2006) Oxidative stress promotes mutant huntingtin aggregation
and mutant huntingtin-dependent cell death by mimicking pro-
teasomal malfunction. Biochem Biophys Res Commun 342:
184 –190.
Graham RK, Deng Y, Slow EJ, Haigh B, Bissada N, Lu G, Pearson J,
Shehadeh J, Bertram L, Murphy Z, Warby SC, Doty CN, Roy S,
Wellington CL, Leavitt BR, Raymond LA, Nicholson DW, Hayden
MR (2006) Cleavage at the caspase-6 site is required for neuronal
dysfunction and degeneration due to mutant huntingtin. Cell
125:1179 –1191.
Gu M, Gash MT, Mann VM, Javoy-Agid F, Cooper JM, Schapira AH
(1996) Mitochondrial defect in Huntington’s disease caudate nu-
cleus. Ann Neurol 39:385–389.
Guidetti P, Charles V, Chen EY, Reddy PH, Kordower JH, Whetsell
WO Jr, Schwarcz R, Tagle DA (2001) Early degenerative changes
in transgenic mice expressing mutant huntingtin involve dendritic
abnormalities but no impairment of mitochondrial energy produc-
tion. Exp Neurol 169:340 –350.
Guidot DM, McCord JM, Wright RM, Repine JE (1993) Absence of
electron transport (Rho 0 state) restores growth of a manga-
nese-superoxide dismutase-deficient Saccharomyces cerevi-
siae in hyperoxia. Evidence for electron transport as a major
source of superoxide generation in vivo. J Biol Chem 268:
26699 –26703.
Gunawardena S, Her LS, Brusch RG, Laymon RA, Niesman IR,
Gordesky-Gold B, Sintasath L, Bonini NM, Goldstein LS (2003)
Disruption of axonal transport by loss of huntingtin or expression of
pathogenic polyQ proteins in Drosophila. Neuron 40:25– 40.
Hanawalt PC, Ford JM, Lloyd DR (2003) Functional characterization of
global genomic DNA repair and its implications for cancer. Mutat
Res 544:107–114.
Handley OJ, Naji JJ, Dunnett SB, Rosser AE (2006) Pharmaceutical,
cellular and genetic therapies for Huntington’s disease. Clin Sci
(Lond) 110:73– 88.
Harjes P, Wanker EE (2003) The hunt for huntingtin function: interac-
tion partners tell many different stories. Trends Biochem Sci 28:
425– 433.
Harman D (1956) Aging: a theory based on free radical and radiation
chemistry. J Gerontol 11:298 –300.
Hazra TK, Hill JW, Izumi T, Mitra S (2001) Multiple DNA glycosylases
for repair of 8-oxoguanine and their potential in vivo functions. Prog
Nucleic Acid Res Mol Biol 68:193–205.
 E. Trushina and C. T. McMurray / Neuroscience 145 (2007) 1233–1248
Hersch SM, Gevorkian S, Marder K, Moskowitz C, Feigin A, Cox M,
Como P, Zimmerman C, Lin M, Zhang L, Ulug AM, Beal MF,
Matson W, Bogdanov M, Ebbel E, Zaleta A, Kaneko Y, Jenkins B,
Hevelone N, Zhang H, Yu H, Schoenfeld D, Ferrante R, Rosas HD
(2006) Creatine in Huntington disease is safe, tolerable, bioavail-
able in brain and reduces serum 8OH2=dG. Neurology 66:
250 –252.
Hilditch-Maguire P, Trettel F, Passani LA, Auerbach A, Persichetti F,
MacDonald ME (2000) Huntingtin: an iron-regulated protein essen-
tial for normal nuclear and perinuclear organelles. Hum Mol Genet
9:2789 –2797.
Hoch FL (1992) Cardiolipins and biomembrane function. Biochim Bio-
phys Acta 1113:71–133.
Horton TM, Graham BH, Corral-Debrinski M, Shoffner JM, Kaufman
AE, Beal MF, Wallace DC (1995) Marked increase in mitochondrial
DNA deletion levels in the cerebral cortex of Huntington’s disease
patients. Neurology 45:1879 –1883.
Huang Z, Hou Q, Cheung NS, Li QT (2006) Neuronal cell death
caused by inhibition of intracellular cholesterol trafficking is
caspase dependent and associated with activation of the mito-
chondrial apoptosis pathway. J Neurochem 97:280 –291.
Huie RE, Padmaja S (1993) The reaction of no with superoxide. Free
Radic Res Commun 18:195–199.
Irwin CC, Wexler NS, Young AB, Ozelius LJ, Penney JB, Shoulson I,
Snodgrass SR, Ramos-Arroyo MA, Sanchez-Ramos J, Pen-
chaszadeh GK et al. (1989) The role of mitochondrial DNA in
Huntington’s disease. J Mol Neurosci 1:129 –136.
Jenkins BG, Rosas HD, Chen YC, Makabe T, Myers R, MacDonald M,
Rosen BR, Beal MF, Koroshetz WJ (1998) 1H NMR spectroscopy
studies of Huntington’s disease: correlations with CAG repeat
numbers. Neurology 50:1357–1365.
Jo E, Darabie AA, Han K, Tandon A, Fraser PE, McLaurin J (2004)
alpha-Synuclein-synaptosomal membrane interactions: implica-
tions for fibrillogenesis. Eur J Biochem 271:3180 –3189.
Karthikeyan G, Lewis LK, Resnick MA (2002) The mitochondrial pro-
tein frataxin prevents nuclear damage. Hum Mol Genet 11:
1351–1362.
Karthikeyan G, Santos JH, Graziewicz MA, Copeland WC, Isaya G,
Van Houten B, Resnick MA (2003) Reduction in frataxin causes
progressive accumulation of mitochondrial damage. Hum Mol
Genet 12:3331–3342.
Kegel KB, Kim M, Sapp E, McIntyre C, Castano JG, Aronin N, DiFiglia
M (2000) Huntingtin expression stimulates endosomal-lysosomal
activity, endosome tubulation, and autophagy. J Neurosci 20:
7268 –7278.
Kegel KB, Meloni AR, Yi Y, Kim YJ, Doyle E, Cuiffo BG, Sapp E, Wang
Y, Qin ZH, Chen JD, Nevins JR, Aronin N, DiFiglia M (2002)
Huntingtin is present in the nucleus, interacts with the transcrip-
tional corepressor C-terminal binding protein, and represses tran-
scription. J Biol Chem 277:7466 –7476.
Kegel KB, Sapp E, Yoder J, Cuiffo B, Sobin L, Kim YJ, Qin ZH, Hayden
MR, Aronin N, Scott DL, Isenberg G, Goldmann WH, DiFiglia M
(2005) Huntingtin associates with acidic phospholipids at the
plasma membrane. J Biol Chem 280:36464 –36473.
Keyer K, Imlay JA (1996) Superoxide accelerates DNA damage by
elevating free-iron levels. Proc Natl Acad Sci U S A 93:13635–
13640.
King MP, Attardi G (1989) Human cells lacking mtDNA: repopulation
with exogenous mitochondria by complementation. Science 246:
500 –503.
Koroshetz WJ, Jenkins BG, Rosen BR, Beal MF (1997) Energy me-
tabolism defects in Huntington’s disease and effects of coenzyme
Q10. Ann Neurol 41:160 –165.
Kuo CF, Mashino T, Fridovich I (1987) alpha, beta-Dihydroxyisovaler-
ate dehydratase. A superoxide-sensitive enzyme. J Biol Chem
262:4724 – 4727.
Kruman II, Wersto RP, Cardozo-Pelaez F, Smilenov L, Chan SL,
Chrest FJ, Emokpae R Jr, Gorospe M, Mattson MP (2004) Cell
1245
cycle activation linked to neuronal cell death initiated by DNA
damage. Neuron 19:549 –561.
Leavitt BR, Guttman JA, Hodgson JG, Kimel GH, Singaraja R, Vogl
AW, Hayden MR (2001) Wild-type huntingtin reduces the cellu-
lar toxicity of mutant huntingtin in vivo. Am J Hum Genet
68:313–324.
Leavitt BR, Raamsdonk JM, Shehadeh J, Fernandes H, Murphy Z,
Graham RK, Wellington CL, Raymond LA, Hayden MR (2006)
Wild-type huntingtin protects neurons from excitotoxicity. J Neuro-
chem 96:1121–1129.
Lee ST, Kim M (2006) Aging and neurodegeneration. Molecular mech-
anisms of neuronal loss in Huntington’s disease. Mech Ageing Dev
127:432– 435.
Li SH, Cheng AL, Zhou H, Lam S, Rao M, Li H, Li XJ (2002) Interaction
of Huntington disease protein with transcriptional activator Sp1.
Mol Cell Biol 22:1277–1287.
Li SH, Gutekunst CA, Hersch SM, Li XJ (1998) Interaction of hunting-
tin-associated protein with dynactin P150Glued. J Neurosci 18:
1261–1269.
Li SH, Li XJ (2004) Huntingtin-protein interactions and the pathogen-
esis of Huntington’s disease. Trends Genet 20:146 –154.
Lill R, Muhlenhoff U (2005) Iron-sulfur-protein biogenesis in eu-
karyotes. Trends Biochem Sci 30:133–141.
Lodi R, Cooper JM, Bradley JL, Manners D, Styles P, Taylor DJ,
Schapira AH (1999) Deficit of in vivo mitochondrial ATP production
in patients with Friedreich ataxia. Proc Natl Acad Sci U S A 96:
11492–11495.
Lodi R, Schapira AH, Manners D, Styles P, Wood NW, Taylor DJ,
Warner TT (2000) Abnormal in vivo skeletal muscle energy me-
tabolism in Huntington’s disease and dentatorubropallidoluysian
atrophy. Ann Neurol 48:72–76.
Lodi R, Tonon C, Calabrese V, Schapira AH (2006) Friedreich’s ataxia:
From disease mechanisms to therapeutic interventions. Antioxid
Redox Signal 8:438 – 443.
MacMillan-Crow LA, Crow JP, Thompson JA (1998) Peroxynitrite-
mediated inactivation of manganese superoxide dismutase in-
volves nitration and oxidation of critical tyrosine residues. Bio-
chemistry 37:1613–1622.
Majumder P, Chattopadhyay B, Mazumder A, Das P, Bhattacharyya
NP (2006) Induction of apoptosis in cells expressing exogenous
Hippi, a molecular partner of huntingtin-interacting protein Hip1.
Neurobiol Dis 22:242–256.
Mann VM, Cooper JM, Javoy-Agid F, Agid Y, Jenner P, Schapira AH
(1990) Mitochondrial function and parental sex effect in Hunting-
ton’s disease. Lancet 336:749.
Manna SK, Zhang HJ, Yan T, Oberley LW, Aggarwal BB (1998)
Overexpression of manganese superoxide dismutase suppresses
tumor necrosis factor-induced apoptosis and activation of nuclear
transcription factor-kappaB and activated protein-1. J Biol Chem
273:13245–13254.
Mao Z, Choo YS, Lesort M (2006) Cystamine and cysteamine prevent
3-NP-induced mitochondrial depolarization of Huntington’s dis-
ease knock-in striatal cells. Eur J Neurosci 23:1701–1710.
Mariani E, Polidori MC, Cherubini A, Mecocci P (2005) Oxidative
stress in brain aging, neurodegenerative and vascular diseases:
an overview. J Chromatogr B Analyt Technol Biomed Life Sci
827:65–75.
Mattson MP (2004) Metal-catalyzed disruption of membrane protein
and lipid signaling in the pathogenesis of neurodegenerative dis-
orders. Ann N Y Acad Sci 1012:37–50.
McCormick ML, Buettner GR, Britigan BE (1998) Endogenous super-
oxide dismutase levels regulate iron-dependent hydroxyl radical
formation in Escherichia coli exposed to hydrogen peroxide.
J Bacteriol 180:622– 625.
McMurray CT (2005) To die or not to die: DNA repair in neurons.
Mutant Res 577:260 –274.
1246 E. Trushina and C. T. McMurray / Neuroscience 145 (2007) 1233–1248
Michalik A, Van Broeckhoven C (2003) Pathogenesis of polyglutamine
disorders: aggregation revisited. Hum Mol Genet 12 (Spec No 2):
R173–R186.
Michiels C, Raes M, Toussaint O, Remacle J (1994) Importance of
Se-glutathione peroxidase, catalase, and Cu/Zn-SOD for cell
survival against oxidative stress. Free Radic Biol Med 17:
235–248.
Milakovic T, Johnson GV (2005) Mitochondrial respiration and ATP
production are significantly impaired in striatal cells expressing
mutant huntingtin. J Biol Chem 280:30773–30782.
Montermini L, Andermann E, Labuda M, Richter A, Pandolfo M, Cav-
alcanti F, Pianese L, Iodice L, Farina G, Monticelli A, Turano M,
Filla A, De Michele G, Cocozza S (1997a) The Friedreich ataxia
GAA triplet repeat: premutation and normal alleles. Hum Mol Genet
6:1261–1266.
Montermini L, Kish SJ, Jiralerspong S, Lamarche JB, Pandolfo M
(1997b) Somatic mosaicism for Friedreich’s ataxia GAA triplet
repeat expansions in the central nervous system. Neurology 49:
606 – 610.
Muhlenhoff U, Richhardt N, Ristow M, Kispal G, Lill R (2002) The yeast
frataxin homolog Yfh1p plays a specific role in the maturation of
cellular Fe/S proteins. Hum Mol Genet 11:2025–2036.
Nucifora FC Jr, Sasaki M, Peters MF, Huang H, Cooper JK, Yamada
M, Takahashi H, Tsuji S, Troncoso J, Dawson VL, Dawson TM,
Ross CA (2001) Interference by huntingtin and atrophin-1 with
cbp-mediated transcription leading to cellular toxicity. Science
291:2423–2428.
Ohshima K, Montermini L, Wells RD, Pandolfo M (1998) Inhibitory
effects of expanded GAA. TTC triplet repeats from intron I of the
Friedreich ataxia gene on transcription and replication in vivo.
J Biol Chem 273:14588 –14595.
Ono M, Kohda H, Kawaguchi T, Ohhira M, Sekiya C, Namiki M,
Takeyasu A, Taniguchi N (1992) Induction of Mn-superoxide dis-
mutase by tumor necrosis factor, interleukin-1 and interleukin-6 in
human hepatoma cells. Biochem Biophys Res Commun 182:
1100 –1107.
Panov AV, Burke JR, Strittmatter WJ, Greenamyre JT (2003) In vitro
effects of polyglutamine tracts on Ca2⫹-dependent depolarization
of rat and human mitochondria: relevance to Huntington’s disease.
Arch Biochem Biophys 410:1– 6.
Panov AV, Gutekunst CA, Leavitt BR, Hayden MR, Burke JR, Stritt-
matter WJ, Greenamyre JT (2002) Early mitochondrial calcium
defects in Huntington’s disease are a direct effect of polyglu-
tamines. Nat Neurosci 5:731–736.
Park S, Gakh O, O’Neill HA, Mangravita A, Nichol H, Ferreira GC,
Isaya G (2003) Yeast frataxin sequentially chaperones and stores
iron by coupling protein assembly with iron oxidation. J Biol Chem
278:31340 –31351.
Perez-Severiano F, Rios C, Segovia J (2000) Striatal oxidative dam-
age parallels the expression of a neurological phenotype in mice
transgenic for the mutation of Huntington’s disease. Brain Res
862:234 –237.
Perutz MF (1999) Glutamine repeats and neurodegenerative
diseases: molecular aspects. Trends Biochem Sci 24:58 – 63.
Piemonte F, Pastore A, Tozzi G, Tagliacozzi D, Santorelli FM, Car-
rozzo R, Casali C, Damiano M, Federici G, Bertini E (2001) Glu-
tathione in blood of patients with Friedreich’s ataxia. Eur J Clin
Invest 31:1007–1011.
Polidori MC, Mecocci P, Browne SE, Senin U, Beal MF (1999) Oxida-
tive damage to mitochondrial DNA in Huntington’s disease parietal
cortex. Neurosci Lett 272:53–56.
Poulsen HE (2005) Oxidative DNA modifications. Exp Toxicol Pathol
57 (Suppl 1):161–169.
Puccio H, Simon D, Cossee M, Criqui-Filipe P, Tiziano F, Melki J,
Hindelang C, Matyas R, Rustin P, Koenig M (2001) Mouse models
for Friedreich ataxia exhibit cardiomyopathy, sensory nerve defect
and Fe-S enzyme deficiency followed by intramitochondrial iron
deposits. Nat Genet 27:181–186.
Puranam KL, Wu G, Strittmatter WJ, Burke JR (2006) Polyglutamine
expansion inhibits respiration by increasing reactive oxygen spe-
cies in isolated mitochondria. Biochem Biophys Res Commun
341:607– 613.
Radi R, Rodriguez M, Castro L, Telleri R (1994) Inhibition of mitochon-
drial electron transport by peroxynitrite. Arch Biochem Biophys
308:89 –95.
Radisky DC, Babcock MC, Kaplan J (1999) The yeast frataxin homo-
logue mediates mitochondrial iron efflux. Evidence for a mitochon-
drial iron cycle. J Biol Chem 274:4497– 4499.
Rao VR, Finkbeiner S (2003) Secrets of a secretase: N-cadherin
proteolysis regulates CBP function. Cell 114:533–535.
Read MA, Whitley MZ, Gupta S, Pierce JW, Best J, Davis RJ, Collins
T (1997) Tumor necrosis factor alpha-induced E-selectin expres-
sion is activated by the nuclear factor-kappaB and c-JUN N-termi-
nal kinase/p38 mitogen-activated protein kinase pathways. J Biol
Chem 272:2753–2761.
Reddy PH (2006) Amyloid precursor protein-mediated free radicals
and oxidative damage: implications for the development and pro-
gression of Alzheimer’s disease. J Neurochem 96:1–13.
Rigamonti D, Bauer JH, De-Fraja C, Conti L, Sipione S, Sciorati C,
Clementi E, Hackam A, Hayden MR, Li Y, Cooper JK, Ross CA,
Govoni S, Vincenz C, Cattaneo E (2000) Wild-type huntingtin
protects from apoptosis upstream of caspase-3. J Neurosci 20:
3705–3713.
Ross CA, Poirier MA (2004) Protein aggregation and neurodegenera-
tive disease. Nat Med 10 (Suppl):S10 –S17.
Rötig A, de Lonlay P, Chretien D, Foury F, Koenig M, Sidi D,
Munnich A, Rustin P (1997) Aconitase and mitochondrial iron-
sulphur protein deficiency in Friedreich ataxia. Nat Genet
17:215–217.
Ruan Q, Lesort M, MacDonald ME, Johnson GV (2004) Striatal cells
from mutant huntingtin knock-in mice are selectively vulnerable to
mitochondrial complex II inhibitor-induced cell death through a
non-apoptotic pathway. Hum Mol Genet 13:669 – 681.
Saft C, Zange J, Andrich J, Muller K, Lindenberg K, Landwehrmeyer B,
Vorgerd M, Kraus PH, Przuntek H, Schols L (2005) Mitochondrial
impairment in patients and asymptomatic mutation carriers of Hun-
tington’s disease. Mov Disord 20:674 – 679.
Sakamoto N, Ohshima K, Montermini L, Pandolfo M, Wells RD (2001)
Sticky DNA, a self-associated complex formed at long GAA*TTC
repeats in intron 1 of the frataxin gene, inhibits transcription. J Biol
Chem 276:27171–27177.
Sawa A (2001) Mechanisms for neuronal cell death and dysfunction in
Huntington’s disease: pathological cross-talk between the nucleus
and the mitochondria? J Mol Med 79:375–381.
Sawa A, Wiegand GW, Cooper J, Margolis RL, Sharp AH, Lawler JF
Jr, Greenamyre JT, Snyder SH, Ross CA (1999) Increased apo-
ptosis of Huntington disease lymphoblasts associated with repeat
length-dependent mitochondrial depolarization. Nat Med 5:1194 –
1198.
Seong IS, Ivanova E, Lee JM, Choo YS, Fossale E, Anderson M,
Gusella JF, Laramie JM, Myers RH, Lesort M, MacDonald ME
(2005) HD CAG repeat implicates a dominant property of hunting-
tin in mitochondrial energy metabolism. Hum Mol Genet 14:
2871–2880.
Schiefer J, Landwehrmeyer GB, Luesse HG, Sprunken A, Puls C,
Milkereit A, Milkereit E, Kosinski CM (2002) Riluzole prolongs
survival time and alters nuclear inclusion formation in a trans-
genic mouse model of Huntington’s disease. Mov Disord
17:748 –757.
Schilling G, Coonfield ML, Ross CA, Borchelt DR (2001) Coenzyme
Q10 and remacemide hydrochloride ameliorate motor deficits in a
Huntington’s disease transgenic mouse model. Neurosci Lett
315:149 –153.
Schulz JB, Dehmer T, Schols L, Mende H, Hardt C, Vorgerd M, Burk
K, Matson W, Dichgans J, Beal MF, Bogdanov MB (2000) Oxida-
 E. Trushina and C. T. McMurray / Neuroscience 145 (2007) 1233–1248
tive stress in patients with Friedreich ataxia. Neurology 55:
1719 –1721.
Shi X, Dong Z, Huang C, Ma W, Liu K, Ye J, Chen F, Leonard SS, Ding
M, Castranova V, Vallyathan V (1999) The role of hydroxyl radical
as a messenger in the activation of nuclear transcription factor
NF-kappaB. Mol Cell Biochem 194:63–70.
Shimohata T, Onodera O, Tsuji S (2000) Interaction of expanded
polyglutamine stretches with nuclear transcription factors leads to
aberrant transcriptional regulation in polyglutamine diseases. Neu-
ropathology 20:326 –333.
Shoichet SA, Baumer AT, Stamenkovic D, Sauer H, Pfeiffer AF, Kahn
CR, Muller-Wieland D, Richter C, Ristow M (2002) Frataxin pro-
motes antioxidant defense in a thiol-dependent manner resulting in
diminished malignant transformation in vitro. Hum Mol Genet
11:815– 821.
Singh KK (2006) Mitochondria damage checkpoint, aging, and cancer.
Ann N Y Acad Sci 1067:182–190.
Sjogren M, Mielke M, Gustafson D, Zandi P, Skoog I (2006) Choles-
terol and Alzheimer’s disease: is there a relation? Mech Ageing
Dev 127:138 –147.
Squitieri F, Cannella M, Sgarbi G, Maglione V, Falleni A, Lenzi P,
Baracca A, Cislaghi G, Saft C, Ragona G, Russo MA, Thompson
LM, Solaini G, Fornai F (2006) Severe ultrastructural mitochondrial
changes in lymphoblasts homozygous for Huntington disease mu-
tation. Mech Ageing Dev 127:217–220.
Steffan JS, Kazantsev A, Spasic-Boskovic O, Greenwald M, Zhu YZ,
Gohler H, Wanker EE, Bates GP, Housman DE, Thompson LM
(2000) The Huntington’s disease protein interacts with p53 and
CREB-binding protein and represses transcription. Proc Natl Acad
Sci U S A 97:6763– 6768.
Sugars KL, Brown R, Cook LJ, Swartz J, Rubinsztein DC (2004)
Decreased cAMP response element-mediated transcription: an
early event in exon 1 and full-length cell models of Huntington’s
disease that contributes to polyglutamine pathogenesis. J Biol
Chem 279:4988 – 4999.
Suopanki J, Gotz C, Lutsch G, Schiller J, Harjes P, Herrmann A,
Wanker EE (2006) Interaction of huntingtin fragments with brain
membranes: clues to early dysfunction in Huntington’s disease.
J Neurochem 96:870 – 884.
Swerdlow RH, Parks JK, Cassarino DS, Shilling AT, Bennett JP Jr,
Harrison MB, Parker WD Jr (1999) Characterization of cybrid cell
lines containing mtDNA from Huntington’s disease patients. Bio-
chem Biophys Res Commun 261:701–704.
Szebenyi G, Morfini GA, Babcock A, Gould M, Selkoe K, Stenoien DL,
Young M, Faber PW, MacDonald ME, McPhaul MJ, Brady ST
(2003) Neuropathogenic forms of huntingtin and androgen recep-
tor inhibit fast axonal transport. Neuron 40:41–52.
Tabrizi SJ, Blamire AM, Manners DN, Rajagopalan B, Styles P, Scha-
pira AH, Warner TT (2005) High-dose creatine therapy for Hun-
tington disease: a 2-year clinical and MRS study. Neurology
64:1655–1656.
Tabrizi SJ, Cleeter MW, Xuereb J, Taanman JW, Cooper JM, Schapira
AH (1999) Biochemical abnormalities and excitotoxicity in Hunting-
ton’s disease brain. Ann Neurol 45:25–32.
Takahashi K, Avissar N, Whitin J, Cohen H (1987) Purification and
characterization of human plasma glutathione peroxidase: a sel-
enoglycoprotein distinct from the known cellular enzyme. Arch
Biochem Biophys 256:677– 686.
Tan G, Chen LS, Lonnerdal B, Gellera C, Taroni FA, Cortopassi GA
(2001) Frataxin expression rescues mitochondrial dysfunctions in
FRDA cells. Hum Mol Genet 10:2099 –2107.
Tang TS, Slow E, Lupu V, Stavrovskaya IG, Sugimori M, Llinas R,
Kristal BS, Hayden MR, Bezprozvanny I (2005) Disturbed Ca2⫹
signaling and apoptosis of medium spiny neurons in Hunting-
ton’s disease. Proc Natl Acad Sci U S A 102:2602–
2607.
Tang TS, Tu H, Chan EY, Maximov A, Wang Z, Wellington CL,
Hayden MR, Bezprozvanny I (2003) Huntingtin and huntingtin-
1247
associated protein 1 influence neuronal calcium signaling
mediated by inositol-(1,4,5) triphosphate receptor type 1. Neu-
ron 39:227–239.
Tang TS, Tu H, Orban PC, Chan EY, Hayden MR, Bezprozvanny I
(2004) HAP1 facilitates effects of mutant huntingtin on inositol
1,4,5-trisphosphate-induced Ca release in primary culture of
striatal medium spiny neurons. Eur J Neurosci 20:1779 –
1787.
The Huntington’s Disease Collaborative Research Group (1993) A
novel gene containing a trinucleotide repeat that is expanded
and unstable on Huntington’s disease chromosomes. Cell 72:
971–983.
Trushina E, Dyer RB, Badger JD 2nd, Ure D, Eide L, Tran DD,
Vrieze BT, Legendre-Guillemin V, McPherson PS, Mandavilli
BS, Van Houten B, Zeitlin S, McNiven M, Aebersold R, Hayden
M, Parisi JE, Seeberg E, Dragatsis I, Doyle K, Bender A, Chacko
C, McMurray CT (2004) Mutant huntingtin impairs axonal traf-
ficking in mammalian neurons in vivo and in vitro. Mol Cell Biol
24:8195– 8209.
Trushina E, Heldebrant MP, Perez-Terzic CM, Bortolon R, Kovtun IV,
Badger JD 2nd, Terzic A, Estevez A, Windebank AJ, Dyer RB, Yao
J, McMurray CT (2003) Microtubule destabilization and nuclear
entry are sequential steps leading to toxicity in Huntington’s dis-
ease. Proc Natl Acad Sci U S A 100:12171–12176.
Tukamoto T, Nukina N, Ide K, Kanazawa I (1997) Huntington’s dis-
ease gene product, huntingtin, associates with microtubules in
vitro. Brain Res Mol Brain Res 51:8 –14.
Valenza M, Rigamonti D, Goffredo D, Zuccato C, Fenu S, Jamot L,
Strand A, Tarditi A, Woodman B, Racchi M, Mariotti C, Di
Donato S, Corsini A, Bates G, Pruss R, Olson JM, Sipione S,
Tartari M, Cattaneo E (2005) Dysfunction of the cholesterol
biosynthetic pathway in Huntington’s disease. J Neurosci
25:9932–9939.
van Dellen A, Grote HE, Hannan AJ (2005) Gene-environment
interactions, neuronal dysfunction and pathological plasticity in
Huntington’s disease. Clin Exp Pharmacol Physiol 32:1007–
1019.
van Loon AP, Pesold-Hurt B, Schatz G (1986) A yeast mutant lacking
mitochondrial manganese-superoxide dismutase is hypersensitive
to oxygen. Proc Natl Acad Sci U S A 83:3820 –3824.
Visner GA, Dougall WC, Wilson JM, Burr IA, Nick HS (1990) Regula-
tion of manganese superoxide dismutase by lipopolysaccharide,
interleukin-1, and tumor necrosis factor. Role in the acute inflam-
matory response. J Biol Chem 265:2856 –2864.
Vorgerd M, Schols L, Hardt C, Ristow M, Epplen JT, Zange J (2000)
Mitochondrial impairment of human muscle in Friedreich ataxia in
vivo. Neuromuscul Disord 10:430 – 435.
Waelter S, Scherzinger E, Hasenbank R, Nordhoff E, Lurz R, Goehler
H, Gauss C, Sathasivam K, Bates GP, Lehrach H, Wanker EE
(2001) The huntingtin interacting protein HIP1 is a clathrin and
alpha-adaptin-binding protein involved in receptor-mediated endo-
cytosis. Hum Mol Genet 10:1807–1817.
White JK, Auerbach W, Duyao MP, Vonsattel JP, Gusella JF, Joyner
AL, MacDonald ME (1997) Huntingtin is required for neurogenesis
and is not impaired by the Huntington’s disease CAG expansion.
Nat Genet 17:404 – 410.
Wilson RB (2003) Frataxin and frataxin deficiency in Friedreich’s
ataxia. J Neurol Sci 207:103–105.
Wong A, Yang J, Cavadini P, Gellera C, Lonnerdal B, Taroni F,
Cortopassi G (1999) The Friedreich’s ataxia mutation confers cel-
lular sensitivity to oxidant stress which is rescued by chelators of
iron and calcium and inhibitors of apoptosis. Hum Mol Genet
8:425– 430.
Wyttenbach A, Swartz J, Kita H, Thykjaer T, Carmichael J, Bradley J,
Brown R, Maxwell M, Schapira A, Orntoft TF, Kato K, Rubinsztein
DC (2001) Polyglutamine expansions cause decreased CRE-me-
diated transcription and early gene expression changes prior to cell
1248 E. Trushina and C. T. McMurray / Neuroscience 145 (2007) 1233–1248

death in an inducible cell model of Huntington’s disease. Hum Mol
Genet 10:1829 –1845.
Yu W, Gong JS, Ko M, Garver WS, Yanagisawa K, Michikawa M
(2005) Altered cholesterol metabolism in Niemann-Pick type C1
mouse brains affects mitochondrial function. J Biol Chem 280:
11731–11739.
Zeron MM, Hansson O, Chen N, Wellington CL, Leavitt BR, Brundin P,
Hayden MR, Raymond LA (2002) Increased sensitivity to N-meth-
yl-D-aspartate receptor-mediated excitotoxicity in a mouse model
of Huntington’s disease. Neuron 33:849 – 860.
Zhou Z, Kang YJ (2000) Cellular and subcellular localization of catalase
in the heart of transgenic mice. J Histochem Cytochem 48:585–594.

(Accepted 27 October 2006)

(Available online 14 February 2007)