Alterations of muscarinic acetylcholine receptors in
atypical Pick’s disease without Pick bodies
T Odawara, K Shiozaki, E Iseki, H Hino, K Kosaka
J Neurol Neurosurg Psychiatry 2003;74:965–967
Background: Atypical Pick’s disease without Pick bodies
is a type of frontotemporal dementia characterised by
semantic dementia and temporal dominant lobar atrophy
with ubiquitinopathy. No neurochemical analyses have
ever been reported in this condition.
Objective: To investigate muscarinic acetylcholine recep-
tors (mAchR) and their subtypes (M1–M4) in atypical Pick’s
Subjects: Five cases of atypical Pick’s disease were stud-
ied. They were compared with nine control cases, 11
cases of Alzheimer’s disease, and seven cases of demen-
tia with Lewy bodies.
Methods: A [3H]quinuclidinyl benzilate (QNB) binding
assay and an immunoprecipitation assay using subtype
specific antisera were used.
Results: The total amount of mAchR in the temporal cortex
was lower in atypical Pick’s disease than in controls or
Alzheimer’s disease cases, but there were no significant
differences between the three groups in the frontal cortex.
In the temporal cortex, there was a smaller proportion of
M1 receptors in atypical Pick’s disease than in the controls
or in the patients with Alzheimer’s disease and dementia
with Lewy bodies. In contrast, the proportion of M2 recep-
tor was higher in atypical Pick’s disease than in the other
Conclusions: Depletion of postsynaptic cholinoreceptive
neurones in the temporal cortex is more severe in atypical
Pick’s disease than in other neurodegenerative dementing
neuronal loss and gliosis in the atrophied regions.1 2Nowa-
days, the use of the term of “Pick’s disease” is restricted to
cases showing lobar atrophy with Pick bodies and tauopathy,
and Pick’s disease is included under the nomenclature of
frontotemporal dementia.3In contrast, cases showing lobar
atrophy without Pick bodies are neuropathologically hetero-
geneous. Some of these cases have clinical features of seman-
tic dementia,which is among the clinical diagnostic criteria of
frontotemporal lobar degeneration (FTLD),4and show tempo-
ral dominant lobar atrophy—hence the name “atypical Pick’s
disease”.5 6Although atypical Pick’s disease shares ubiquitin-
opathy with the dementia that occurs with motor neurone
disease,5 7it is differentiated from that disorder by absence of
clinical features of amyotrophic lateral sclerosis and preserva-
tion of lower motor neurones.6Atypical Pick’s disease is
considered to be a temporal dominant type of fronto-temporal
Neocortical cholinergic deficits have been extensively
investigated in some forms of neurodegenerative dementia in
relation to memory disturbance, especially in Alzheimer’s dis-
ick’s disease is a rare form of neurodegenerative
dementia, neuropathologically characterised by circum-
scribed frontotemporal lobe atrophy as well as by severe
ease and dementia with Lewy bodies.8–10However, investiga-
tion into the cholinergic systems in fronto-temporal dementia
has been inadequate, and have only been undertaken in the
frontal lobe degeneration type of dementia and in the Pick
type with Pick bodies.11 12Systemic studies of muscarinic ace-
tylcholine receptors (mAchR) in atypical Pick’s disease have
never been done.
In this study,we estimated the total amounts of mAchR and
the levels of the mAchR subtypes in atypical Pick’s disease.
We examined brains from five cases of atypical Pick’s disease
(mean (SD) age, 71.4 (6.4) years, range 64 to 81; mean post-
mortem delay to specimen retrieval 3.4 (1.4) hours (table 1).
The clinical,neuropathological,and neurochemical features of
these cases have been described in detail previously.6In sum-
mary, all cases showed semantic dementia and had a clinical
diagnosis of FTLD.There was no family history of this disorder
in any case. Neuropathologically, all cases showed lobar atro-
phy of the frontal and temporal lobes with temporal lobe
dominance. There was moderate to severe degeneration with
neuronal loss and gliosis in the affected cerebral cortices and
subcortical nuclei. In the pyramidal tract, the right cerebral
peduncle was degenerated but the hypoglossal nucleus and
the anterior horn of the spinal cord were preserved.
Immunohistochemical analyses in all cases showed ubiq-
dendritic inclusions in the atrophied cortex. Immunoblots of
samples from the cases did not show detectable amounts of
The dissected brain tissues were immediately frozen in liq-
uid nitrogen, and then stored at −80°C until used.
For comparison, we studied 11 cases of Alzheimer’s disease
(mean age 78.4 (8.7) years, range 63 to 86; mean postmortem
delay 2.6 (1.9) hours), seven cases of dementia with Lewy
Abbreviations: ChAT, choline acetyltransferase; FTLD, frontotemporal
lobar degeneration; mAchR, muscarinic acetylcholine receptors; QNB,
Summary of the cases of atypical Pick’s
Case 1 Case 2Case 3Case 4 Case 5
Age at onset (y)
Brain weight (g)
Cited from ref 6.
F, frontal lobe; T, temporal lobe; Y, years.
bodies (mean age 77.9 (6.9) years, range 68 to 86; mean post-
mortem delay 4.4 (1.5) hours), and nine controls with no
notable neuropathological findings (mean age 84.9 (5.3)
years, range 75 to 91; mean postmortem delay 5.7 (5.6)
hours).All cases selected for comparative study,except for two
of the cases in the control group, had been used previously in
The methods used have been described previously in
detail.10In brief, the amounts of total mAchR in all cases were
estimated at the specific binding sites of [3H]quinuclidinyl
benzilate (QNB) in membrane preparations from the frontal
and temporal cortices. The samples of frontal and temporal
cortices were prepared, respectively, from the frontal pole and
the rostral parts of the temporal lobes that were most severely
affected. The amounts of proteins were measured using an
assay kit from Bio-Rad (Hercules,California,USA).An immu-
noprecipitation assay using specific antisera against AchR M1,
M2, M3, and M4 was done to estimate the levels of mAchR
subtypes.Diverse antisera were raised against GST fusion pro-
teins incorporating the putative third intracellular loop.
One way analysis of variance followed by the least significant
difference test was used to compare the values of total QNB
binding and the proportions of subtypes M1 to M4. Statistical
evaluation was done using the SPSS package.
The amounts of mAchR at the [3H]QNB binding sites in mem-
brane preparations ranged from 0.8 to 2.5 pmol/mg protein
(frontal cortex) and from 0.8 to 2.1 pmol/mg protein (tempo-
ral cortex). In the temporal cortex, values for the [3H]QNB
binding sites in atypical Pick’s disease were significantly lower
than in the control or Alzheimer’s disease groups,although no
substantial differences were not found between atypical Pick’s
disease and dementia with Lewy bodies. In contrast, there
were no significant differences in the binding sites in the
frontal cortex among the various groups.
Membrane preparations from all cases were used for
precipitation with each of the antisera against M1, M2, M3,
and M4 receptors. The proportions of [3H] QNB–receptor
complex precipitated with each of the antisera as solubilised
complexes were estimated (table 2). In the frontal cortex, the
proportion of [3H]QNB–receptor complex precipitated with
anti-M3 antiserum was significantly lower in atypical Pick’s
disease than in Alzheimer’s disease. In the temporal cortex,
the proportion of anti-M1 complex was significantly lower in
atypical Pick’s disease than in the other groups. In contrast,
the proportion of anti-M2 complex was higher in atypical
Pick’s disease than in the other groups. Comparisons between
the other three groups are omitted here, as they have been
dealt with elsewhere.10
There have been few systematic neurochemical studies on
cholinergic alterations in cases of fronto-temporal dementia.
Wood et al reported that the activity of choline acetyltrans-
ferase (ChAT) and the numbers of mAchR were unchanged in
the cerebral cortex in three cases of Pick’s disease with Pick
bodies.11Both Hansen et al and Francis et al found that ChAT
activity was similar to control values, and that the numbers of
mAchR were decreased in the temporal cortex in all of seven
cases of Pick’s disease with Pick bodies, although the decrease
was less than in Alzheimer’s disease.12 13Procter et al showed
that the numbers of mAchR did not fall in 10 cases of Pick’s
disease with Pick bodies or in six cases of fronto-temporal
dementia of the frontal lobe degeneration type, but did fall in
nine cases of Alzheimer’s disease in the frontal, temporal, and
parietal cortices.14In summary, previous studies on Pick’s dis-
ease with Pick bodies and the frontal lobe degeneration type of
fronto-temporal dementia have shown that numbers of
mAchR are either preserved or reduced to some extent,
although ChAT activity—which indicates presynaptic cholin-
ergic innervation—is preserved.
The incidence of atypical Pick’s disease is similar to that of
Pick’s disease with Pick bodies in Japan.15Our results show
that the decrease in mAchR was more severe in the temporal
cortex in atypical Pick’s disease than in controls or Alzheimer’s
disease cases. However, no substantial differences were found
in the total amounts of mAchR between atypical Pick’s disease
and dementia with Lewy bodies. This may accounted for the
finding that the neocortical cholinergic deficit is more exten-
sive in dementia with Lewy bodies than in Alzheimer’s
In the frontal cortex, the proportion of M3 receptors was
lower in atypical Pick’s disease than in Alzheimer’s disease.
This result may reflect the relatively higher proportion of M2
receptors in atypical Pick’s disease, but this is not known for
certain. In the temporal cortex, there was a decrease in the
proportion of M1 receptors, the most abundant muscarinic
acetylcholine receptor in the cerebral cortex,16while the
proportion of M2 receptors, which exist at presynapses of the
subtype specific antisera
Immunoprecipitation of muscarinic acetylcholine receptor subtypes using
Proportions of precipitated mAchR (%)
M1 M2 M3M4
Data are mean (SD) data from a representative experiment. The density of mAchR was estimated as specific
binding with [3H]QNB, and ranged from 800 to 2500 fmol/mg membrane protein. The proportions of
specially precipitated mAchR subtypes were estimated as the differences in the amounts of bound [3H]QNB
precipitated with specific antiserum and those precipitated with non-immune serum.
*p<0.05, **p<0.01; *** p<0.001 v controls; †p<0.05, ††p<0.01, †††p<0.0015 v AD; ‡p<0.01,
‡‡p<0.001 v DLB.
AD, Alzheimer’s disease; aPD, atypical Pick’s disease; DLB, dementia with Lewy bodies; mAchR, muscarinic
acetylcholine receptors; QNB, [3H]quinuclidinyl benzilate.
966Odawara, Shiozaki, Iseki, et al
cholinergic system,17increased. The lower level of M1
receptors in atypical Pick’s disease may reflect the greater
severity of neuronal losses in the temporal cortex in this con-
dition in comparison with Alzheimer’s disease or dementia
with Lewy bodies, because all cases of atypical Pick’s disease
showed severe neuronal loss with gliosis in the anterior
portions of the temporal lobes.6Moreover, the higher level of
M2 in atypical Pick’s disease raises the possibility that the
presynaptic M2 receptors are upregulated as a result of the
loss of cholinoreceptive neurones in the temporal cortex,
although it is not possible to determine the absolute
proportion of M1 to M4 receptors by our immunoprecipitation
method. These findings suggest that projecting cholinergic
neurones are relatively preserved, in contrast to the loss of
cholinoreceptive neurones in the temporal cortex, although
ChAT was not estimated in this study.A previous neuropatho-
which the cholinergic neurones project to the temporal cortex,
was mildly degenerated in Pick’s disease, including atypical
Pick’s disease,18supporting our hypothesis.
Acetylcholine esterase inhibitors are used in the treatment
of Alzheimer’s disease,and are also of potential use in demen-
tia with Lewy bodies10and vascular dementia. Unfortunately,
our study suggests that these agents may be ineffective in
treating cholinergic deficiency in patients suffering from
atypical Pick’s disease, because the cholinoreceptive neurones
appear to be more severely affected than the projecting presy-
naptic cholinergic neurones.
This study was supported in part by grants-in-aid for scientific
research from the Japan Ministry of Education, Science, Sports and
T Odawara, E Iseki, H Hino, K Kosaka, Department of Psychiatry,
Yokohama City University School of Medicine, Yokohama, Japan
K Shiozaki, Yokohama Maioka Hospital, Yokohama
Competing interests: none declared
Correspondence to: Dr Toshinari Odawara, Department of Psychiatry,
Yokohama City University School of Medicine, 3-9 Fukuura,
Kanazawa-ku, Yokohama 236-0004, Japan;
Received 27 November 2002
In revised form 21 February 2003
Accepted 9 March 2003
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