Abstract Mutations in the PARK7 gene DJ-1 are associ-
ated with recessive hereditary Parkinson’s disease (PD).
Fibrillar inclusions of α-synuclein comprise the neu-
ropathological hallmarks of PD and related Lewy body
diseases as well as multiple system atrophy (MSA). More-
over, neuronal and glial inclusions containing tau have
been observed in α-synucleinopathy patients. Using a col-
lection of antibodies against DJ-1, we have performed a
comprehensive investigation of DJ-1 in α-synucleino-
pathies and tauopathies. DJ-1 was abundantly expressed
in reactive astrocytes of patients with neurodegenerative
diseases. Likewise, DJ-1 antiserum immunostained reac-
tive astrocytes that became abundant with disease pro-
gression in the brain stem of transgenic mice expressing
mutant [A30P]α-synuclein. Human Lewy bodies as well
as Lewy body-like inclusions in the α-synuclein trans-
genic mice were DJ-1 negative. Neuronal tau inclusions
were DJ-1 immunopositive in Pick’s disease (PiD), corti-
cobasal degeneration (CBD), progressive supranuclear
palsy (PSP), and Alzheimer’s disease. In addition, we
found DJ-1-immunopositive glial inclusions in CBD, PSP
and MSA. Biochemical extraction experiments revealed
the specific presence of insoluble, modified DJ-1 in PiD
and MSA. Our results suggest that DJ-1 is up-regulated in
reactive astrocytes as well as in neuronal and glial cells
with specific α-synucleinopathy and tauopathy.
Keywords DJ-1 · Synuclein · Tau · Multiple system
atrophy · Pick’s disease
Introduction
DJ-1 is a ubiquitously expressed protein whose function is
poorly understood [2, 7, 19, 29, 31, 43]. Previous research
on DJ-1 focused on its role in male reproductive tissue,
where DJ-1 is thought to modulate the transcriptional ac-
tivity of the androgen receptor by interacting with the reg-
ulatory proteins PIASxα and DJBP [35, 39]. An essential
role of DJ-1 in brain emerged with the discovery of DJ-1
mutations underlying autosomal-recessive hereditary Par-
kinson’s disease (PD) [2]. DJ-1 deletions and point muta-
tions were subsequently found worldwide [1, 18]. Both
deletion and point mutations in the DJ-1 gene led to loss
of functional protein, in the case of the well-studied
[L166P]DJ-1 due to accelerated protein degradation [17,
24, 26, 30, 36]. The deficit that accounts for dopaminergic
deficiency in DJ-1 mutation bearers [8] is not known. Pos-
sible roles of DJ-1 including oxygen radical scavenging or
chaperone activity are currently being discussed [7].
Here we studied the pathophysiological properties of
DJ-1 in patients with PD and neuropathologically related
α-synucleinopathies [9, 14, 20], including dementia with
Lewy bodies (DLB), neurodegeneration with brain iron ac-
cumulation type 1 (NBIA-1, formerly known as Hallervor-
den-Spatz disease), and multiple system atrophy (MSA).
Given the emerging interrelation of α-synucleinopathy and
tauopathy [12], we included diseases with tau inclusion
body formation: Alzheimer’s disease (AD), Pick’s disease
(PiD), corticobasal degeneration (CBD), progressive
supranuclear palsy (PSP) and argyrophilic grains disease
(AGD) [5, 42]. Immunohistochemistry on postmortem tis-
sue revealed strong expression of DJ-1 in reactive astro-
cytes in disease-affected regions in all patients. Although
intraneuronal α-synuclein (αSYN) inclusions were not
stained with anti-DJ-1, (oligo)dendroglial cytoplasmic in-
clusions (GCIs) from MSA patients did contain DJ-1.
Likewise, we found that oligodendroglial and astrocytic tau
inclusions in PSP, CBD and AGD as well as neuronal tau
inclusions (some neurofibrillary tangles and most Pick
bodies) were immunoreactive with anti-DJ-1. Biochemical
analysis confirmed the presence of sarcosyl-insoluble,
Manuela Neumann · Veronika Müller · Karin Görner ·
Hans A. Kretzschmar · Christian Haass · Philipp J. Kahle
Pathological properties of the Parkinson’s disease-associated protein DJ-1
in α-synucleinopathies and tauopathies:
relevance for multiple system atrophy and Pick’s disease
Acta Neuropathol (2004) 107 : 489–496
DOI 10.1007/s00401-004-0834-2
Received: 12 December 2003 / Revised: 21 January 2004 / Accepted: 21 January 2004 / Published online: 26 February 2004
REGULAR PAPER
M. Neumann · H. A. Kretzschmar
Institute of Neuropathology,
Ludwig Maximilians University of Munich, Munich, Germany
V. Müller · K. Görner · C. Haass · P. J. Kahle (✉)
Laboratory of Alzheimer’s and Parkinson’s Disease Research,
Department of Biochemistry,
Ludwig Maximilians University of Munich,
Schillerstrasse 44, 80336 Munich, Germany
Tel.: +49-89-5996480, Fax: +49-89-5996415,
e-mail: pkahle@pbm.med.uni-muenchen.de
© Springer-Verlag 2004

modified DJ-1 in the fibrillar fractions prepared from MSA
and PiD brains. The observed up-regulation of DJ-1 might
reflect a protective response. Early-onset PD in patients
with complete loss of DJ-1 indicates that the nigrostriatal
dopamine system is particularly dependent on DJ-1.
Materials and methods
Primary antibodies
Three antibodies against DJ-1 were used: polyclonal antisera against
raised against synthetic peptides 138–151 (SN1130) and 174–189
(SN1132) of human DJ-1 [38], and mouse monoclonal antibody
3E8 (Biozol; Eching, Germany) raised against amino acids 1–187
of the human DJ-1 protein [31]. Rat monoclonal 15G7 was raised
against amino acids 116–131 of human αSYN [21]. Mouse mono-
clonal anti-tau purchased from Innogenetics (Zwijndrecht, Bel-
gium) recognized tau phosphorylated at serine-202 and threo-
nine-205 (AT8) [16] and threonine-231 (AT180) [15]. Mouse mono-
clonal antibody Tau-2 recognizing phosphorylated and non-phos-
phorylated tau was purchased from Sigma (St. Louis, MO).
Immunohistochemistry
Sections (4 µm) were cut from formalin-fixed, paraffin-embedded
tissue blocks from representative brain regions from selected cases
(Table 1). In addition, sections were analyzed from transgenic
mice expressing human αSYN with the A30P mutation under the
control of the Thy1 promoter [21, 34].
To enhance immunoreactivity, tissue sections were boiled in
10 mM citrate buffer, pH 6.0, in a microwave oven (five times for
3 min each). To block nonspecific antibody binding sites, tissue
sections were incubated in 2% bovine serum albumin/0.01% (v/v)
Triton X-100 for 30 min at room temperature. Incubation with pri-
mary antibody was performed for 1.5 h at room temperature.
Working dilutions for primary antibodies were: AT8, 1:100;
AT180, 1:200; Tau-2, 1:250; 3E8, 1:500; SN1130, 1:200; SN1132,
1:300; 15G7, 1:10. Antibody binding was detected using the alka-
line phosphatase/anti-alkaline phosphatase system (DAKO, Ham-
burg, Germany), following the manufacturer’s instructions. New
fuchsin was used as the chromogen.
Double immunolabeling of DJ-1 and αSYN was performed
with two different fluorophore-conjugated secondary antibodies.
After incubation with the first primary antibody against DJ-1
(SN1130 and 3E8) for 1.5 h, sections were incubated for 30 min
with tetramethyl rhodamine isothiocyanate-conjugated goat anti-
rabbit IgG and goat anti-mouse IgG (Dianova, Hamburg, Ger-
many), respectively. The second primary antibody 15G7 against
αSYN was then added for 1.5 h and detected using fluorescein-
conjugated goat-anti rat IgG (DAKO). This antibody was pre-ab-
sorbed with either rabbit or mouse IgG (DAKO) to minimize cross-
reaction with the first primary antibody. Sections were analyzed
with a Leica TCS-NT confocal laser-scanning microscope.
490
Table 1 Demographic and
neuropathological information.
Neuritic plaques and neurofi-
brillary tangles were scored ac-
cording to the CERAD [27]
and the Braak and Braak [3]
classification, respectively. LB
distribution was scored accord-
ing to the consensus guidelines
for the pathological diagnosis
of DLB [25] (LB Lewy body,
AD Alzheimer’s disease,
PiD Pick’s disease, CBD corti-
cobasal degeneration, PSP pro-
gressive supranuclear palsy,
AGD argyrophilic grains dis-
ease, PD Parkinson’s disease,
DLB dementia with LBs,
NBIA-1 neurodegeneration
with brain iron accumulation
type 1, MSA multiple system
atrophy, PM postmortem,
n.a. not available)
No. Diagnosis Age (years) Sex PM delay CERAD Braak stage LBs
1 AD 73 F <24 C VI –
2 AD 81 F 40 C VI –
3 AD 88 M 4 C V –
4 AD 79 M 48 C VI –
5 AD 87 M 26 C VI Amygdala
6 AD 88 F 72 C VI Amygdala
7 PiD 66 F 4 0 0 –
8 PiD 54 M n.a. 0 0 –
9 CBD 77 F 12 0 I –
10 CBD 62 M n.a. 0 0 –
11 PSP 67 M 40 0 0 –
12 PSP 63 F 11 0 0 –
13 PSP 63 M 34 0 II –
14 AGD 78 F 39 0 III –
15 AGD 58 F 48 0 II –
16 PD 77 F 48 0 I Limbic
17 PD 75 M 15 C III Neocortical
18 PD 83 F n.a. A IV Neocortical
19 PD 83 M 29 A II Neocortical
20 DLB 80 M 7 0 II Neocortical
21 DLB 77 F 14 C V Neocortical
22 NBIA-1 27 F n.a. 0 0 Neocortical
23 MSA 64 F 9 0 I –
24 MSA 64 M 9 0 0 –
25 MSA 58 F 24 0 I –
26 MSA 66 M 20 0 0 –
27 Infarct 87 M 26 B II –
28 Infarct 78 F n.a. 0 0 –
29 Control 85 M 25 B III –
30 Control 54 M n.a. 0 I –
31 Control 67 M 8 0 0 –