Dopamine

receptors

disease
PHILIP

and transporters

and schizophrenia
SEEMAN,t

Departments

AND

of Pharmacology4

HYMAN

B. NIZNIK1

and Psychiatryt,

University

ABSTRACT

The loss of midbrain dopamine

in Parkinsons disease
is accompanied
by a matching
loss in the dopamine
transporter
and a rise in the Dl and D2 receptor densities. This is found in the brain putamen and caudate
tissues from unmedicated
patients, and may account
for the good early clinical response to L-dopa. Longterm L-dopa treatment
reverts the receptor densities
toward normal levels. Positron emission tomography
(PET) data and in vitro data generally concur. In
schizophrenia
the density of the dopamine
transporter
as well as that of the Dl dopamine
receptor is normal.
The D2 receptor
density, however,
is consistently
elevated in postmortem
brain putamen
and caudate
nucleus, even in tissues from neuroleptic-free
or drugnaive patients. Three sets of PET and single photon
emission computed
tomography
(SPECT)
data support the postmortem
findings. Early evidence indicating abnormal D2 structure as well as a reduced link between Dl and D2 warrant a detailed study of the genes
for these two receptors in schizophrenia.SEEMAN, P.;
NIZNIK,
H. B. Dopamine receptors and transporters
in
Parkinsons
disease and schizophrenia.
FASEB j 4:
2737-2744;
1990.
Key
tor

Words:
L-dopa
caudate nucleus

postmortem tissue

dopamine

recep-

schizophrenia

PARKINSONS
DISEASE
AND schizophrenia
each have abnormal
dopamine
function
in addition
to many other
disabling
features.
For example,
the loss of midbrain
dopamine
neurons
(1) results in the rigidity and tremor
of patients
with Parkinsons
disease,
these signs and
symptoms
being
alleviated
by replacement
therapy
with L-dopa1
(2). On the other hand,
schizophrenia,
with its disordered
thinking
patterns,
hallucinations,
and delusions,
suggests
a state of hyperdopamine
activity, as these symptoms
are usually alleviated
by antipsychotic
drugs that specifically
block dopamine
D2
receptors
(3).
In the early stages of both diseases,
the patients
are
supersensitive
to dopamine-mimetic
medications,
with
Parkinson
patients
improving
on L-dopa
(4) but the

0892-6638/90/0004-2737/$01

in Parkinsons

.50. FASEB

of Toronto, Toronto, Ontario,

Canada

M5S

1A8

schizophrenic
patients
becoming
worse (see references
in refs 5 and 6). Parkinson
patients
who are overmedicated
with L-dopa also have psychotic
symptoms,
including
hallucinations,
which go away when the dose
of L-dopa is lowered.
These clinical supersensitivities
to dopamine
in both
types of patients
may be attributed
to the elevated
densities of dopamine
D2 receptors
found
in studies
of
postmortem
tissue. The purpose of this review is to examine
the abnormalities
of dopamine
receptors
and
transporters
that may account
for some of the clinical
features.
DOPAMINE
DISEASE

TRANSPORTER

IN

PARKINSONS

Although
the number
of doparnine-containing
cell bodies in the Parkinson
midbrain
(substantia
nigra)
is
reduced
by 60-86%
of age-matched
control tissues (1),
the loss of dopamine
in the nerve terminals
in the striaturn is even greater,
amounting
to a 90-99%
loss in the
putamen
(2, 7-9) and a 58-95%
loss in the caudate
nucleus
(2, 7-9).
Until now it has been puzzling
that the dopamine
transporter,
which is located
on the nerve terminals,
falls by only 22-60%,
whereas
the dopamine
content
drops by more than 90%. For example,
Fig. 1 (top) IIlustrates
that the dopamine
transporter,
as labeled
by
[11C]nomifensine,
was 22%
below
control;
similar
reductions
of 22-60%
in the striatal dopamine
transporter
are also shown
in Fig.
1 (top)
for either
[3H]GBR
12935 or [3H]cocaine.
These
reductions,
however,
do not correspond
to the reductions
of
95-99%
in the dopamine
content
of postmortem
Parkinsons diseased
putamen
(2).
This discrepancy
has recently been resolved by H. B.
Niznik,
E. L. Fogel, F. F. Fassos, and P. Seeman
(unpublished
results),
who found that the dopamine
transport sites (defined as [3H]GBR
12935 binding displaced

Abbreviations:
L-dopa, levo-dihydroxyphenylalanine;
[25I]FAPP,
1-(2-[bis-(4-fluorophenyl)-methoxy]ethyl)-4-(2-[4-azido-3-[
25I]iodophenyl]ethyl)piperazine;
PET,
positron
emission
tomography;
SPECT,
single photon
emission
computed
tomography;
5, striatum;
P, putamen;
C, caudate
nucleus;
[25I]azidoNAPS,
N-(p-azido-m[251]iodophenethyl)spiperone;
[3H]ADTN,
( )6,7-dihydroxy-2aminotetralin.

2737

PARKINSONS

by 1 tM mazindol)
were virtually
absent in the Parkinson putamen
after the tissue had been partly purified
by using a wheat-germ
agglutinin
column.
An example
is shown in Fig. 2 (top) which illustrates
that [3H]GBR
12935 binds to two sites, as revealed
by the inhibition
of binding
by mazindol.
That is, one component
of
[3H]GBR
binding
was inhibited
by about
30 nM
mazindol,
whereas
the
other
component
(the
piperazine-binding
site) was inhibited
by approximately
50,000 nM mazindol.
The top of Fig. 2 indicates that
[3H]GBR
12935 binds
to approximately
equal amounts
of these two sites in the human
putamen. [The piperazine-binding
site has recently
been
identified
as a P450-like
protein (14), and therefore
may
be important
in causing
or preventing
Parkinsons

100

Ow
-I.-.

zo

I-

0
6

160

21

20

16

Dl RECEPTORS
NOL-DOPA

g
0
z
0

disease.]

Because
the dopamine
transporter
is a surface
membrane-associated
protein,
surface
glycoprotein
is
attached,
permitting
the separation
of the transporter
from the intracellular
membranes
by passing the solubilized tissue through
a wheat-germ
agglutinin
column.
This separation
has been done for the results shown at
the bottom
of Fig. 2. These data reveal the complete
absence
of the dopamine
transporter
(that is, the portion of [3H]GBR
binding
displaced
by 1 tM mazindol)
in the Parkinson
tissue, compared
with normal
human

100

200

tissue

C.,

g
10o
0

CD

Ifl

CD

I
I

Figure

OOOCOO
-

1. Parkinsons
disease
exhibits
a reduced
or absent
dopamine transporter
(top) associated
with the loss of the dopaminecontaining
nigral neurons.
In untreated
Parkinsons
disease,
the Dl
and D2 receptors are elevated,
only to return
to normal
upon Ldopa therapy.
100% is defined
by the value for the non-Parkinson
control tissues or patients. The number at the base of each column
indicates
the number
of diseased
tissues or of patients
studied.
5,
striatum;
P, putamen;
C, caudate
nucleus.
Dopamine transporter:
Tedroff S: [Cj()
Nomifensine
(ref 10); Pearce: 5: [H]GBR 12935;
3 nM (ref II); Janowsky:
C: [3H]GBR
12935; 0.9 nM (ref 12);
Maloteaux:
C: [3H]GBR 12935; 0.3 nM (ref 9); Hirai: P: [3HJGBR
12935; 0.5 nM (ref 13); Schoemaker: P: [3H]cocaine,
10 nM (ref 8);
Niznik: Method of Fig. 2; 2 nM [3HJGBR
12935. C: 36% remaining; P: 0% remaining.
Dl receptors using /3HJSCH 23390: Raisman:
P: 1 nM (ref 16); Seeman: S: Bm,,density
(ref 17); Cortes: S: 1 nM (ref
18) (slices);
Pimoule: P: 0.4 nM (ref 7); Costello: P and C: (ref 18a;
slices) Pierot: P and C: 2 nM (ref 18b). D2 receptors using [3H]spiperone
unless otherwise stated: Lee: 5: [3H]Haloperidol,
1.5 nM (ref 19); Rinne:
P: 0.8 nM (ref 20); Guttman:
S: Bm,, density
(ref 21); Seeman: Bm,,
density
(ref 17); Pimoule: P: 1 nM (ref 7); Rutgers: S:[C}methylspiperone
(ref 22); Perlmutter: C: [8Flspiperone
(ref 23); Hagglund:
S: [uC]methylspiperone
(ref 24); EARLY
indicates
Stage I patients; Reisine: 0.8 nM (ref 25); Ba/cobsa: 0.5 nM (ref 26); Cortes: 1 nM

13H]CV 205 502 (ref

2738

Vol. 4

18) (slices).

July 1990

(control.)

An example
of the absence
of the dopamine
transporter in Parkinsons
disease is seen in Fig. 3, where the
transporter
has been selectively
labeled with [251]FAPP
(ref. 15; H. B. Niznik
et al., unpublished
results).
Figure
3 illustrates
the
covalent
attachment
of
[1251]FAPP to tissue incubated
with [1251]FAPP and exposed
to light.
The
attached
[251]FAPP
migrates
together
with the dopamine
transporter
protein
and
identifies
the apparent
molecular
weight of the transporter
as approximately
68,000.
(Although
there are
two dark bands in each lane of the gel, only the band
at 68,000 was specifically
displaced
by 1 tM mazindol.)
The 68,000 band was present
in the human
putamen
from control
brain,
Alzheimer-diseased
brain,
and
schizophrenia
brain, but was completely
absent in the
Parkinsons
diseased
brain.

DOPAMINE
DISEASE

RECEPTORS

IN PARKINSONS

Consistent
with the degeneration
and loss of dopamine
neurons
in Parkinsons
disease,
the density
of postsynaptic
dopamine
receptors
increases
in association
with the developing
denervation
dopamine
supersensitivity of the patient
(or the animal or the brain striatal
tissue).
There
are currently
two major classes of dopamine
receptors:
Dl and D2 (see refs in ref. 6). Dl is the dopamine receptor
that stimulates
adenylate
cyclase and is
selectively
labeled by [3H]SCH
23390. D2 is the dopamine receptor
that inhibits
adenylate
cyclase and is
selectively
blocked
and labeled
by [3H]antipsychotic
drugs
such as [3H]spiperone,
[3H]haloperidol,
and
[3H]raclopride.
Each of these receptors
can exist in a

The FASEB Journal

SEEMAN AND

NIZNIK

studies
did not report
the densities
of dopamine
receptors
but
only
the striatum/cerebellum
ratios,
which
range between
1.5 and 3.4. Such low signal/background

PET ratios need to be increased

to clarify this important issue.
It is possible that the elevated densities
of Dl and D2
dopamine
receptors
may account
for the Parkinsons
diseased
patients
clinical
supersensitivity
to dopamine
agonists
(4, 19). With
prolonged
dopamine
agonist
therapy
(L-dopa,
bromocriptine),
the densities
of Dl
and D2 receptors
generally
return
to or approach
normal levels,
as summarized
in Fig. 1 (right
side, middle
and
bottom
panels).
This
agonist-induced
downregulation
has suggested
that a clinical
L-dopa
holiday
be used
to rehypersensitize
patients
who become
less
sensitive
to L-dopa
(27), wherein
the receptors
would
be permitted
to up-regulate
during
the agonist
holiday.
Although
much
attention
has been
given
to dopamine
receptors
in Parkinsons
disease,
other
receptors
have also been measured
and usually
found
to be normal (see ref. 28 for a review),
unless
the receptor
is lo-

S
0
0

0.
ci

E
0

z
0
U,

C,

cated on the degenerating

nigra neuron
(possibly
neurotensin
and y-aminobutyric
acid binding
sites).
PET
results
usually
concur
with the postmortem
findings.
For example,
monkeys
with unilaterally
le-

Putamen
MAZINDOL, moles/LIter

GBR 12909

Figure 2. The dopamine transporter

was absent in the homogenate
of the Parkinson putamen that had been lectin-purified
by wheatgerm
agglutinin
chromatography.
Specific
binding
of [3H]GBR
12935 to the dopamine
transporter
was defined
by that which was
inhibited
by 1 I4M mazindol.
The agglutinin
removed
the piperazinc binding Site, a P450-like
protein
(14).

highor low-affinity
state for dopamine.
For example,
the dissociation
constant
of dopamine
for the highaffinity
state of Dl is 0.7 nM, whereas
that for D2 is 5
nM.
The
dissociation
constants
of dopamine
for the
low-affinity
states
of Dl or D2 are both
around
3000

nM.
The
disease
tients
weeks
or D2

elevations
of Dl and D2 densities
in Parkinsons
are summarized
in Fig. 1. In tissues from pawho died
without
receiving
L-dopa
in the last
of life, the binding
of a radioactive
ligand to Dl
was

control
kinsons

elevated

20-60%

above

the

of PET

ample,

Rutgers

(positron
emission
tomography).
For exet al. (22) did not find elevated
binding
of[C]methylspiperone
to D2 receptors
in six patients,
whereas
Perlmutter
et al. (23) found a 40% increase
in
[8F]spiperone
binding
in one patient
with Parkinsons
disease
(see Fig.
1, bottom).
These
latter
two PET

DOPAMINE

RECEPTORS, PARKINSONS,

AND

200

69-i

92

46

neurological

tissue. The situation

is less clear in living Pardiseased
patients
who have been studied
by

means

Mr

SCH IZOPH REN IA

30-

Mazindol
Figure
3. The dopamine
transporter
was absent
(indicated
by arrow) in the Parkinsons
diseased
putamen,
as detected by photoaffinity
labeling
with [251]FAPP
(15; H. B. Niznik et a!., unpublished results).

2739

sioned
dopamine
neurons
reveal a loss of dopamine
transporters
(29) with an elevated
level of D2 receptors
in vivo (29; M. Guttman,
personal
communication),
in
agreement
with postmortem
data (Fig. 1). The PET-

DOPAMINE

observed
binding
of
rises in the Parkinsonian

to D2 receptors
some time before

normal
in
schizophrenia,

to the dopamine

four different
types of transporter-labeling
[3H]GBR
12935, [3H]cocaine,
[1251]FAPP,
zindol.
The density
of Dl dopamine
receptors
has also been
found to be normal in schizophrenia
subjects,
as shown
for 99 different
striatal tissues in Fig. 4 (middle panel),
except for 8 tissues studied by Hess et al. (37). Although
Dl is most reliably measured
by using [3H]SCH
23390,
the specific conditions
used by the authors in Fig. 4 (middle) permitted
the measurement
of Dl. Whereas
the Dl
density
may be normal
in schizophrenia
patients
in
general,
it is possible that the link between
Dl and D2
receptors
is abnormal
(see below).

[C]raclopride
animal
(29)

the fall in [C]nomifensine

transporter
(29).
This

binding

early
rise
in [C]raclopride
binding
may be partly
explained
by both a rise in the
D2 density
as well as the removal
of endogenous
dopamine
that normally
competes
for [3H]raclopride
bind-

ing (30).
An important
difference,
however,
vitro (Fig.
1) and PET data is that
[3H]SCH
23390 (to Dl) was increased
tem striata
premortem

from

PET
ealed a reduced
striata (31).

unilaterally
lesioned
data on these same

uptake

of [11C]SCH

between
the in
the binding
of
in the postmormonkeys,
but the
monkeys
had rev23390 into their

RECEPTORS

AND

TRANSPORTER

IN SCHIZOPHRENIA
The density

of dopamine

transporter

postmortem
as summarized

sites appears

to be
with
4 for
ligands:
and [3H]ma-

tissues
of
patients
in the top of Fig.

SCHIZOPHRENIA
100

Ow
I-,-

ZO

o.
00

Figure

I-

transporter

4. Schizophrenia
reveals normal amounts
of dopamine
(top) and Dl receptors (middle). D2 receptors, however,

are consistently elevated in postmortem striata, even if the patients

had not been taking neuroleptics. Of four in vivo studies, three revealed statistically significant elevations () in binding of the radioligand to D2 receptors
in unmedicated
schizophrenics.
S, stria0

tum; P, putamen;
C, caudate.
Dopamine transporter, using [3HJGBR
12935, unless stated otherwise: Hirai: P: 0.5 nM (ref 13); Pearce: S: 3 nM
(ref 1!); Czudek: C: Bma,, density (ref 32); Niznik: P: [25IIFAPP (unpublished
results); Pimoule: P: 10 nM [3Hjcocaine (ref 7);Joyce: 5: 15
nM [3H]mazindol; slices (ref 33). Dl receptorsusing [3HJSCH 23390,
unless stated otherwise: Cross: Drug free for more than 1 year; 2 nM

100

I-

[3H]flupenthixol

with

100 nM domperidone

to occlude

D2 (ref 34);

Seeman: P: [3H]dopamine
Bm,, (35);
Cross: C: [3HIADTN
(ref 36);
Hess: C: Bma,, (ref 37); Joyce: 1 nM; slices; never medicated (ref 33);
Pimoule: P: 0.4 nM (ref 7); Seeman 87: 5: Bmax (ref 17); Reynolds: C:
Bm,, (ref 38). D2 receptors using [3HJspiperone, unless stated otherwise.
Reynolds: P: Bm,, (ref 39); Kornhuber: P: Bmax (ref 40); Seeman: P or
C: [3H]raclopride
Bmax (ref 41); Lee (Spip): P: 1 nM (ref 42); Mackay
82: C: Bm,, (ref 43); Reisine (B): P: Bm,, (ref 25); Pimoule: P: 1 nM
(ref. 7); Lee 1982: P: Bm,, (ref 44); Owen 1978: P: 0.8 nM (ref 45);
Reisine: P: 0.1 nM (ref 25); Hess: P: Bma,, (ref 37); Lee (Halo): P: 2
nM [3H]haloperidol (ref 42); Seeman 84: P: Bmax (ref 46); Seeman 87:
S: Bm,, (ref 17); Owen 1983: P: Bmx (ref 47); Lee (B): P: Bmax (ref
42); Toru: P: Bm,, (ref 48); Mita: C: Bma,, (ref 49). D2 receptors (patients had been off neuroleptics) using [3H]spipeivne: Mackay 80: C: B,,,,,,
(ref 50); Kornhuber: P: Bmu; off drugs more than 10 days (ref 40);
Cross: P: Bm,,,; off drugs more than 1 year (ref 36): Toru. P: B,,,,,; off
drugs more than 40 days (ref 48); Joyce: S slices; I nM; never on
drugs (ref 33); Mita: C: Bm,,,; off drugs more than 60 days (ref 49).
D2 by PET or SPECT (patients unmedicated or never treated): Farde: P:

200
c.1

0
I0

0
100

[C}Raclopride (ref 51) (Data for left putamen

only; 18 patients).
Martinot: [76Br]Bromospiperone;
striatum;
chronic patients.
The subchronic patients
(4)
were significantly
different
from controls (52, 53);
Crawley: [77Br]Bromospiperone
SPECT
of striatum; off drugs more
than 6 months (ref 54); Wong: [1 C]Methylspiperone;
caudate
(ref 55).

2740

Vol. 4

July 1990

The FASEB Journal

SEEMAN AND

NIZNIK

The density
of D2 receptors
is consistently
elevated
in postmortem
striatal tissues (Fig. 4, bottom).
Although
Mackay
et al. (50) indicated
that the elevated
D2 density is induced entirely by the administration
of neuroleptic drugs during
the patients lifetime,
marked
D2 elevations have been found by Joyce et al. (33) and by T. J.
Crows group (34) in tissues from schizophrenic
patients
who had never taken neuroleptics.
Furthermore,
although
Kornhuber
et al. (40) reported
that postmortem
D2 densities were not elevated in tissues from patients
who had
not received
neuroleptics
for 10 days or more before
death, their findings are not supported
by others (33, 36,
48, 49), as shown in Fig. 4.
The long-term
administration
of neuroleptics
to patients increases
the density of D2 dopamine
receptors
in
postmortem
human
tissues by only 25%. For example,
the top panel of Fig. 5 indicates
that the mode for the
D2 density in the striata from 122 different
individuals
was 12.9 pmol of receptor per gram of original tissue (17).
Virtually
the same value (13.4 pmol/g) was found in the
tissues from Alzheimer
patients
who had not received
any neuroleptics
during
their lifetime
(Fig. 5, third
panel). Those Alzheimer
patients who received neuroleptics premortem
subsequently
revealed a D2 density that
was approximately
25% higher (16.6 pmol/g)
than the
nonneuroleptic
Alzheimer
tissues (compare
third and
fourth panels in Fig. 5).
In schizophrenia,
however,
at least half of the brain
striata revealed
D2 densities
between
22 and 40 pmol/g
(Fig. 5, second panel from top), values that were double
to triple
those measured
in control
tissues (17). Thus,
these high D2 densities
cannot be accounted
for by the
neuroleptic-induced
increase of 25% in D2 seen patients
with Alzheimers
disease or Huntingtons
disease (results
not shown; see ref. 17). (Huntingtons
diseased
patients
take neuroleptic
doses equivalent
to those taken
by
schizophrenic
patients).
PET measurements
of D2 in drug-free
(52-54)
and
drug-naive
(51, 55) patients
are still unclear. Statistically
significant
elevations
in D2 were found in 10 patients
by
Wong et al. using PET (55), in 12 patients
by Crawley
et al. using SPECT
(53), and in 6 subchronic
patients
by Martinot
et al. using PET (52, 53). However,
Martinot et al. (52, 53) did not find a significant
elevation
in
the striatum/cerebellum
ratio when
averaging
eight
chronic

patients.

Using

[11C]raclopride,

Farde

et al. (51)

found that neuroleptic-naive

schizophrenic
patients
revealed a statistically
significant
elevation
of 3 pmol/g in
D2 in the left putamen,
compared
with the right putamen in the same patients.
Farde et al. (51) also found
that the schizophrenic
left putamen
was elevated by about
3 pmol/g
compared
with control
volunteers,
but the
difference
was not statistically
significant.
In many PET studies,
including
that of Farde et al.
(51), the cerebellum
is used as a baseline
for nonspecific
binding
of the radioisotope.
However,
as the nonspecific
binding
of [3H]raclopride
by the cerebellum
is 20-50%
lower than that for nonspecific
binding
in the striatum
(56), the absolute
densities
for D2 calculated
by Farde
et al. (51) (values of about 28 pmol/g)
are much higher

DOPAMINE

RECEPTORS, PARKINSONS,

AND

SCHIZOPHRENIA

NEUROLEPTICS

ON D2

CONTROL

STR IATA
12.9 0.2 pmol/g

SCHIZOPHRENIA

ALZHEIM ERS
WITHOUT
NEUROLEPTICS
13.4 0.6 pmol/g

ALZH

ElM ERS

WITH

NEUROLEPTICS

16.6 0.6pmol/g

D2

DENSITY pmol/g

Figure 5. Each box indicates a different postmortem human striatum D2 density, as measured
using [3Hjspiperone
(Bm,,,;
centrifugation
method (17). The D2 densities
in postmortem
striata
from
schizophrenic
patients
exhibit
a bimodal
pattern,
with half the

values being two to three times higher than normal. Although

neuroleptic-treated
Alzheimer
patients or Huntington patients (17)
revealed elevated D2, the magnitude and pattern were different
from that for schizophrenia (adapted from ref. 17).

the postmortem
values of 13 to 14 pmol/g (wherein
nonspecific
binding
is defined by the striatum).
Thus,
although
D2 is elevated
in the left schizophrenic
putamen by 11% when the cerebellum
is used as baseline,
this elevation
may be closer to 22% when using another
nonspecific
brain region (such as striatum
or frontal cortex) as baseline
(56).
than

2741

DOPAMINE

RECEPTOR

FUNCTION

IN SCHIZOPHRENIA

STRUCTURE

% INHIBITION

AND

(OF THE DOPAMINE

EFFECT)

BY SCH 23390

Bmax [KD] 0 10 20 30 40 50 60708090100%

I

CONTROLS

The selective
block of D2 by antipsychotic
drugs, and
the elevated
D2 densities
in drug-free
and drug-naive
schizophrenic
tissues, suggest that the protein structure
of D2 in schizophrenic
patients
may be abnormal.
This
receptor
abnormality
need not be constant,
because
schizophrenia
is a disease that waxes and wanes during
its natural
course.
It is possible,
for example,
that the
two forms of D2 (see references
in refs 57 and 58), as
well as additional
variants yet to be found, may alter during the psychotic
illness.
The molecular
weight of D2 is approximately
44,000
after the glycocalyx
has been removed
from the receptor, as noted in Fig. 6. Further
enzymatic
digestion
of
the 44,000 protein
by papain
and subsequent
gel electrophoresis
in a second direction
(shown in Fig. 6) revealed a unique peptide fragment
in tissues with high D2
density
from schizophrenic
subjects.
Such results were
not seen in tissues digested
with other enzymes,
such as
protease
from Staphylococcus aureus (N. H. Bzowej, H. B.
Niznik,
and P. Seeman,
unpublished
results). The data
in Fig. 6 were obtained
from D2 receptors
that were first
photoaffinity-labeled
by [125I]azidoNAPS.
Therefore,
the
data shown in Fig. 6 suggest that there may be an abnormal
structure
of D2 in schizophrenia
that warrants
an examination
of the D2 gene.
Finally, there is evidence
for an abnormal
or reduced
link between
Dl and D2 in schizophrenia
(41). The
Dl-D2 link is tested as follows. The addition
of 400 nM
dopamine
in vitro reduces
the apparent
D2 density, as
labeled by [3Hjraclopride
(41). This is because dopamine
occupies
the high-affinity
state of D2 very tightly with
a dissociation
constant
of 5 nM, as mentioned
previously. The attached
dopamine
added at the high concentration
of 400
nM
prevents
some
of the
[3H]raclopride
from occupying
D2, despite a dissociation constant
of 2 nM for [3Hjraclopride
for the D2

Control

[I,
1st

C
0
C,)
C
0

Schizophrenic
Schizophrenic
[High Density] [Normal Density]

44K

44K

44K

IT
#{163}

Figure 6. The peptide map of D2 receptors

from schizophrenic
tissue reveals an additional
fragment compared
with control tissue.

The tissue homogenate

had been photolabeled
with [251]azidoNAPS,
deglycosylated,
chromatographed
in 1 dimension,
the 44000 mol wt
protein
cut Out, digested
with papain,
and chromatographed
in a
second direction
(N. H. Bzowej,
H. B. Niznik, and P. Seeman,
unpublished
results).

2742

Vol. 4

July 1990

9.9 12.0
6.5 11.6
[11.0

*11.1

18.4 5.8
*14.5
14.4
7.7

5.7

8.5

8.8

I
I

1.9
1.7

7.0 0.8

11.5

2.6
2.0
1.3

I
I

19.4
11.3

8.5

5.6
6.9
10.0

7.0
9.1

12.6

*19.3 4.8
* 17.2 3.7
*8.4 2.0
*12.3
* 9.4 5.8
* 21.7 6.6
*21.0
7.4
*14.21 3.6
*14.4 9.1
*20.4 4.7
10.5 2.4
* 11.3
8.8
*10.6 2.9
*15.3
4.0

SCHIZOPHRENIA
I
F
F

tI
ii
F
F

ti
I

Figure

7. More than half the tissues from schizophrenic

patients
revealed
a reduced
or absent
link between
Dl and D2 dopamine
receptors.
Each bar indicates
the percent
inhibition
by SCH 23390
(of the dopamine-induced
depression
of [3H]raclopride
density).
Details
in ref 41.

receptor. Hence, the dopamine

added exogenously
lowers
the apparent
binding
of [3H]raclopride.
However,
when
human
tissue is preincubated
with 150 nM SCH 23390
(which selectively
acts on the Dl receptor),
the addition
of exogenous
dopamine
no longer reduces
the binding
of the D2-selective
[3H]raclopride.
The
molecular
mechanism
of this D1-D2 link (explained
more fully in
ref 41) appears
to be based on the common
f3-y subunit
of the G proteins
associated
with the Dl and D2
receptors.
The D1-D2 link appears to be reduced in postmortem
schizophrenic
tissue, as SCH 23390 is not capable
of
blocking
the action of exogenous
dopamine
in this tissue (41) (summarized
in Fig. 7). Inhibition
of the dopamine effect by SCH 23390 in control and Parkinsons
diseased tissues was 100% but in many of the schizophrenic
tissues, no inhibition
of the dopamine
effect could be obtained.
The
abnormality
in this receptor-receptor
link,
however,
is not confined
to tissues in schizophrenic
patients, as it was also seen in tissues from patients
who
died with Huntingtons
psychosis
(41). Such data suggest that there may be an abnormal
structure
to the Dl
receptor
in psychosis
and/or
schizophrenia.
If so, a
reduced
influence
from Dl would allow excessive numbers of D2 receptors
to remain in the high-affinity
state.
As there is reason to think that the high-affinity
state is

The FASEB Journal

SEEMAN AND

NIZNIK

state of D2, such a preponderance

of highwith the clinical conof schizophrenia.

distinct [3HIGBR-12935
ophys. 276, 424-432

the functional

.affinity D2 states may be associated

E!i

dition
Partly

supported

by the

and the Ontario

Mental
late J. Aubrey Medland,

Medical

Health

Research

Foundation.

Council

Scientist

of the Ontario

Ministry

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F, and Agid,
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Eur. j Pharmacol. 113, 467-468

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