REV IEW

Dopamine receptors:
from structure to behavior
Stuart C. Sealfon and C. Warren O l a n o w

The responses obtained with drugs that act at dopamine receptors depend on the spectrum of
receptors stimulated, the pattern of stimulation and the neuronal signal-transduction pathways
that are activated. In the absence of drugs that reliably discriminate between the various cloned
receptors, elucidating the role of these receptors has largely relied on molecular genetic approaches
that include expression of genes for receptors in cell lines and manipulation of this expression in
animal models. Connecting molecular events that occur consequent to receptor stimulation with
the resulting physiological effects entails bridging a complex network of interactions. This article
reviews the current understanding of the molecular, cellular and systemic consequences of
activation of the different dopamine receptors.
Trends Neurosci. (2000) 23, $34-$40

HE EVIDENCE that lack of dopamine (DA)-receptor
T stimulation contributes to Parkinson's disease (PD)
symptoms, and that receptor activation has a beneficial
effect on these symptoms appears incontrovertible.
The principal effects of L-dOpa occur after its conversion
to DA via stimulation of various DA receptors. Direct
acting DA-receptor agonists that do not require meta-
bolic conversion and act directly at DA-receptor sites
have similarly been shown to provide benefit in PD.
However, the receptor stimulation profile and resulting
clinical benefits of L-dopa and DA-receptor agonists are
not identical. For example, clinical benefits are greater
with L-dOpa, whereas DA-receptor agonists are less likely
to induce motor complications such as dyskinesia and
motor fluctuations. In order to understand better how
L-dopa and the various DA-receptor agonists modulate
behavioral responses differentially, the results of their
actions on specific DA receptors must be understood.
Considerable progress has been made in elucidating
the anatomy, neurochemistry, physiology, pharmacol-
ogy and molecular biology of the dopaminergic system
and its receptors ~-3. DA receptors are members of a large
family of structurally related receptors, the rhodopsin-
like G-protein-coupled-receptor (GPCR) superfamily.
With the recognition that there are multiple DA recep-
tors, it has become evident that the response to a DA-
receptor agonist is likely to depend on its relative activ-
ity at these molecularly identified receptors. However,
Stuart C. Sealfbn determining the relationship between receptor activation
and and physiological response is difficult. The responses that
C. Warren Olanow ultimately occur after receptor activation result from
are at the Dept of modulation of a complex network of signaling molecules
Neurology and and neuronal circuits. Resolving these issues is compli-
Stuart C. Sealfon cated by a lack of drugs that are selective for the cloned
is also at the receptor subtypes, and by the existence of both immedi-
Fishberg Center for ate and long-lasting effects in response to receptor
Neurobiology, stimulation. For example, treatment with L-dOpais associ-
Mount Sinai ated with immediate motor benefits and with late-
School of onset motor complications that presumably reflect
Medicine, drug-induced plastic changes in the response network.
New Yolk, The ultimate response to an agonist is determined by
N Y 10O29, USA. the specific receptors that are selected and by the degree
to which signal-transduction pathways are activated.
Mapping this response pattern is likely to be complex:
drugs can activate more than one receptor, receptors
can activate more than one signal-transduction path-
way and different agonists acting at the same receptor
can differentially affect these signaling pathways. Thus,
the effects of an agonist are determined not merely by
the identities of the receptors activated and the degree
of activation, but also by the specific pattern of signal
transduction that is elicited.
Dopamine-receptor subtypes
Dopamine D1 and D2 receptors were initially charac-
terized based on differences in ligand selectivity, and
positive (D1) or negative (D2) coupling to adenylate
cyclase4. Molecular cloning revealed the existence of
five (D1-D5) DA-receptor subtypes. The cloned D1 and
D5 receptors are pharmacologically Dl-like, and the
genes for the D2-D4 receptors encode a D2-1ike family
of receptors z. The rat D2 receptor was the first DA
receptor to be cloned, being isolated by its homology
to the [~2-adrenergic receptor s. Like all DA receptors
cloned to date, it has seven hydrophobic domains that
constitute predominantly c~-helical membrane-span-
ning segments. The distribution of D2 receptor mRNA
has been studied by in situ hybridization. Highest lev-
els are observed in the striatum, nucleus accumbens
and olfactory tubercle, as well as in dopaminergic neur-
ons in the midbrain, where they presumably generate
autoreceptors. The gene for the D2 receptor also gives
rise to two receptor isoforms, through alternative exon
splicing, that differ in the presence (D2L) or absence
(D2S) of a 29 amino-acid segment in the third cyto-
plasmic loop 2. These isoforms show differences in
G-protein coupling, regional distribution and sequestra-
tion rate 6-8. In contrast to cloned D2-1ike receptors,
cloned D1 receptors have a short third cytoplasmic loop.
Highest levels of D1 receptor mRNA are found in the
striatum, nucleus accumbens and olfactory tubercle.
The D3 receptor has about 75% similarity to the D2
receptor in the transmembrane domains, but differs in
its pharmacological profile, signal-transduction coupling

$34 71NSVol. 23, No. 10 (Suppl.), 2000 0 [66-2236•00/$ see front matter © 2000 Elsevier Science Ltd. All rights reserved. Pll: $1471-1931(00)00025-2
 S . C . Sealfon and C . W . Olanow - Dopamine receptors: f r o m structure t o behavior REV~EW

and distribution 9. D3-receptor mRNA levels are lower conformations of the receptor. As DA-receptor agonists
in the striatum and higher in nucleus accumbens t h a n stabilize different active conformations, they might direct
D2-receptor mRNA. Alternative transcripts of the D3 the receptor stimulus to different G proteins. This has
receptor have been identified, but only mouse alternative been termed 'stimulus trafficking '2° . Alterations in the
transcripts encode functional receptor isoforms. normal pattern of tonic and phasic DA-receptor acti-
The sequence of the D4 receptor has approximately vation 21, as might occur with d o p a m i n e depletion and
50% similarity to D2 receptors in t r a n s m e m b r a n e with stimulation of the denervated receptor by therapy
domains 1°. Levels of mRNA are highest in frontal cortex, with exogenous DA-like agents, might also lead to altered
amygdala, olfactory bulb and hypothalamus. The h u m a n signaling and behavioral patterns.
D4 receptor is highly polymorphic, with a 16 amino- Evidence for stimulus trafficking arises from the rever-
acid-repeat segment (two to ten times) in the third sal of the relative efficacies of agonists in stimulating
cytoplasmic loop domain. A specific repeat n u m b e r has different signaling pathways via the same receptor.
been correlated with novelty-seeking personality traits n, Reversal of p o t e n c y or efficacy has been identified with
and increased delusions in psychotic individuals are several GPCRs. For example, pituitary adenylate cyclase-
associated with a high n u m b e r of repeat segments 12. activating peptide (PACAP)-27 is more p o t e n t t h a n
In rodents, DS-receptor mRNA is primarily located in PACAP-38 in stimulating cAMP p r o d u c t i o n via the
the olfactory tubercle, h i p p o c a m p u s and mammillary PACAP receptor, but less p o t e n t in stimulating inositol
nucleus, and n o t in the striatum. phosphate accumulation 2z. The efficacies of the 5-HT1a-
D1 and D2 receptors in the striatum are primarily receptor agonists rauwolscine and ipsapirone are reversed
f o u n d on m e d i u m spiny neurons where t h e y serve to for activation of Gj~z compared with G~ 3 (Ref. 23).
modulate glutamate-mediated activity ~a. Dl-receptor- Differential stimulation of the inositol phosphate and
bearing neurons contain substance P and dynorphin, arachidonic-acid second-messenger system has also been
and give rise to n e u r o n s comprising the direct striato- demonstrated for a series of partial agonists at the
pallidal pathway. D2-receptor-bearing neurons contain h u m a n 5-HT2A and 5-HT2c receptors z4. Although signal
enkephalin and give rise to neurons that influence the trafficking has n o t yet been described for DA-receptor
pallidum by way of the indirect pathway. Recent agonists, the presence of this p h e n o m e n o n in structurally
observations suggest that this model m i g h t be overly related receptors suggests that they are likely to have
simplistic. There is evidence for extensive collateraliz- similar properties. Thus, the behavioral effects obtained
ation of striatofugal axons TM,the presence of D2 receptors with DA-receptor agonists result from the conformational
on striatal interneurons that is increased after a lesion states that are stabilized by the agonist complexing with
of dopaminergic neurons ~s, and co-localization of D1 its specific receptor and the pattern of signal-transduction
and D2 receptors o n striatal m e d i u m spiny neurons 16. responses that are elicited by these conformational states.
Furthermore, there is evidence of gap junctions that Several groups have probed the conformational
provide anatomical connections between axons of changes that occur during GPCR activation. Site-directed
striatal neurons, and these are markedly increased under spin labeling has been used to measure the relative
conditions of dopaminergic denervation or therapy m o v e m e n t of helix 3 and helix 6 side chains of
with DA-like agents ~7,18. These observations are n o t rhodopsin during activation zs. The results demonstrate
accounted for in the present model. that receptor activation is associated with displacement
of the cytoplasmic ends of the two helices and a counter-
Molecular mechanisms of action
clockwise rotation of helix 6. In support of the idea
The rhodopsin-like GPCRs, which include DA recep- that m o v e m e n t s of these helices contribute to receptor
tors, comprise several h u n d r e d h o m o l o g o u s proteins activation, rhodopsin activation is blocked w h e n the
that transduce an extracellular signal into intracellular m o v e m e n t of helices 3 and 6 is restricted by either
G-protein activation. Members of this hepta-helical disulfide crosslinking 2s or engineered Zn2÷-binding sites 26.
protein family are identified by the presence of a series Similarly, a constitutively active 62-adrenergic receptor
of conserved amino-acid motifs within their trans- has been shown to manifest increased accessibility of a
m e m b r a n e segments ~9. Activation of GPCRs leads to helix-6 cysteine for chemical modification. This finding
altered receptor conformation. The activated receptor is consistent with counterclockwise rotation of this helix
induces GDP-GTP exchange in the c~subunit of a hetero- during activation. Site-specific environment-sensitive
trimeric G protein, which causes dissociation of the e~ fluorescent labeling studies also suggest that agonists
and [3y G-protein subunits and subsequent modulation induce conformational changes of helix 3 and helix 6,
of various intracellular effector proteins. The simplest consistent with a model of GPCR activation in which
model for receptor activity presupposes that GPCRs exist helix 6 rotates and displaces from helix 3 (Fig. 1) 2s.
in either an active or inactive conformational state. The first crystal structure of the GPCR rhodopsin has
Agonists exert their effects by preferentially b i n d i n g to recently been reported z7. This crystal structure validates
and stabilizing the active conformation. However, there the insight into the structure and function of these
is n o a priori reason w h y there should be only a single receptors that have been developed t h r o u g h integration
active receptor conformation. Proteins are capable of of various experimental and computational approaches.
assuming a n u m b e r of different conformations that are Cryo-electron microscopy studies of rhodopsin have
distributed according to an 'energy landscape '2°. It is identified the orientation of the t r a n s m e m b r a n e helix
probable that there are several different active receptor domain bundle of these receptors 28. Various approaches
conformational states, with some being more stable have been used to map the binding site of DA receptors,
than others. Agonists stabilize active conformations, and including substituted cysteine-scanning mutagenesis,
different agonists m i g h t stabilize different populations site-directed mutagenesis and receptor chimeras. The
of active conformations. M a n y receptors are k n o w n to binding pocket of DA receptors, c o m m o n to all neuro-
activate multiple G proteins, and different G proteins transmitter GPCRs, lies within the transmembrane helix
might recognize and interact with different active d o m a i n (Fig. 2). There is evidence that GPCRs can form

TINS Vol. 23, No. 10 (Suppl.), 2000 S35

REv-VT~ S.C. S e a l f o n a n d C . W . O l a n o w - Dopamine receptors: from structure to behavior
I Activation
Basal ganglia, Parkinson's disease and levodopa therapy: TINS supplement
Fig. 1. Movement of helix 6 during G-protein-coupled-receptor acti-
vation. A model of rhodopsin is shown, viewed from the cytoplasmic
face. This model is based on electron-density maps and spin-labeling
studies. Selectedhelix 3 and 6 side chains are indicated by colored circles.
During activation (bottom), helix 6 tilts and rotates with the cytoplasmic
side of helix 6 being displaced from helix 3. This altered protein confor-
mation leads to interactions with G proteins and activation of intraceflular
signaling pathways. Adapted from Ref. 25.
homo- and heterodimers ~°, and gel electrophoresis
provides evidence of DA-receptor dimers in cell lines and
brain samples 31,3z. GPCRs can also aggregate in vivo,
and these aggregations can be functionally significant.
Using fluorescence resonance energy transfer assays, it
can be shown that the somatostatin ssts receptor and the
D2 receptor form heterodimers that modulate recep-
tor signaling :33. For example, signal transduction of a
coupling-deficient mutant somatostatin receptor can
be restored by co-expression of the gene for the mutant
receptor with a D2 receptor, suggesting that the somato-
statin receptor can signal through the D2 receptor.
Functional interactions between DA receptors and ligand-
gated channels have also been reported. The C-terminal
domain of the D5 receptor can interact with the second
intracellular loop of the GABAA-receptor y2 subunit, so
that each receptor inhibits the function of the other 34.
Molecular genetic approachesto understanding
DA-receptor function
DA-receptor agonists and antagonists might not
reliably distinguish the various cloned DA receptors.
As a result, several laboratories have turned to genetic
manipulations to investigate the role of specific DA
receptors. Mice with genetically induced lesions that
target the D1, D2, D3 and D4 receptors have been
$36 TINS Vol. 23, No. 10 (Suppl.), 2000
characterized. Antisense sequences for the receptor,
using an oligonucleotide or expression vector, can also
assess decreased function of a genetically identified
receptor. Transgenic receptor overexpression mouse
models can be used to assess the effects of gain of
function. In this approach, additional copies of the gene
of interest are introduced into the pronucleus of a fer-
tilized egg, and the effects of the expression of the
transgene are studied. To date, an overexpression
transgenic model has only been developed for the gene
for the D1 receptor 3s. There are several caveats that must
be considered when evaluating genetic approaches to
the study of DA-receptor function. In gene lesion and
transgenic approaches, animals develop with the gen-
etic abnormality. Accordingly, the phenotype might
be influenced by developmental adaptations to this
perturbation as well as the background strain in which
the genetic alteration is studied. These have both been
proposed to influence the phenotype of D2-receptor-
deficient mice3% The approach used to introduce the
genetic lesion can also lead to variations in pheno-
type. In some models, the protein is completely elimi-
nated, whereas in others, a truncated protein can be
produced that does not form binding sites, but might
retain other nonreceptor functions.
In transgenic mice, the gene expressed is incorpo-
rated into the genome in a random location. In order to
exclude the possibility that inadvertent inactivation of
another gene at the site of incorporation contributes
to the phenotype, it is necessary to study multiple,
independently generated lines. In addition, the location
and level of expression of the receptor gene is deter-
mined both by the promoter sequence used for target-
ing and the site of transgene incorporation. The lack
of promoters that predictably target DA receptors to the
desired regions has limited this approach. Hybrid arrest
approaches attempt to ablate the receptor through the
introduction of antisense oligonucleotides or vectors.
This approach allows the animal to develop normally,
but the reduction in receptor level is incomplete.
Furthermore, this approach is restricted by physical
limitations in delivery of antisense into target cells.
Although these limitations need to be borne in mind,
genetic approaches have provided important insights
into the role of DA receptors.
Lesions in the gene for the D1 receptor have been
reported to induce deficits in movement initiation and
reaction to external stimuli in one model :<3s, but not
in another 39. Mice with a genetically lesioned D1
receptor have altered sensitization to amphetamine
and do not show the progressive increase in motor
responses seen in control animal¢ °. These results sug-
gest that D1 is the predominant receptor involved in
sensitization to amphetamine. Deficits in spatial learn-
ing and reduced locomotor responses to repeat cocaine
injections have also been reported in this modeP 1,42.
Accompanying these behavioral changes, Dl-receptor
null mice exhibit a reduction in substance P and
enkephalin synthesis, increased immunostaining for the
GluR1 glutamate-receptor subunit, and decreased
staining for the NR1 subunit of the NMDA receptor 43.
Induction of expression of Fos and Junb, and regulation
of dynorphin by cocaine and amphetamine are absent.
Interestingly, neurophysiological responses and cel-
lular morphology in the neostriatum are normal in
this mouse. However, dopamine does not potentiate
NMDA-receptor-mediated electrophysiological responses
 S.C. Sealfon and C . W . Olanow - Dopamine receptors: from structure to behavior
in neostriatal slices in Dl-receptor-deficient mice 1:~.In
addition, dopamine-mediated inhibition of glutamate-
induced firing is lost in nucleus accumbens neurons.
The effects of D2-receptor agonists on glutamate-
induced activation in the nucleus accumbens are also
absent, even t h o u g h D2 receptors are functional in
the Dl-receptor-deficient mice, as evidenced by the
preserved induction of cataplexy and expression of
striatal Fos and ]un by D2-receptor antagonists.
The effects of overexpressing the gene for the D1
receptor have been investigated in two lines of trans-
genic mice 3s, Increased Dl-receptor binding was most
marked in cortical areas. A full Dl-receptor agonist
caused a striking suppression of locomotion, in contrast
to the dose-dependent increase in locomotor activity
seen in wild-type mice. Dl-receptor-agonist-induced
rearing and climbing behaviors were suppressed, but
the transgenic animals performed as well as control
mice on rotarod testing, indicating that sensorimotor
coordination was unaffected. These results show that
altering the levels of D1 receptor can reverse the effects
of Dl-receptor agonism on locomotor initiation and
rearing. This raises the possibility that D1 receptors in
different locations might contribute to both suppression
and stimulation of movement initiation.
Two lines of mice with genetically lesioned D2
receptors have been developed 36'44. Impaired locomotor
function has been observed in each, although there was
some disparity in the degree of deficit. D2-receptor-
lesioned mice also showed an abnormal pattern of long-
term depression, with synaptic depression being replaced
by potentiation l:~. These findings implicate the loss of
D2-receptor activation in the motor deficits observed in
PD. Antisense approaches have also been applied to study
D2-receptor function in vivo. Bilateral injection into the
striata of mice of an expression plasmid that contains
the antisense D2-receptor sequence led to cataplexy
and inhibition of stereotypy induced by D2-receptor
agonist 4s. Depletion of postsynaptic D2 receptors in the
rat striatum with antisense oligonucleotide diminished
stereotypic sniffing in response to a high dose of apo-
morphine. Vacuous chewing, which is induced by low-
dose apomorphine, was unaffected, suggesting the
involvement of other receptors or presynaptic D2 recep-
tors in this response. Unilateral depletion of presynaptic
D2 receptors, using antisense oligonucleotides injected
into rat substantia nigra, induced contralateral rotation
in response to cocaine. These results suggest that
presynaptic D2 receptors contribute to the locomotor
responses observed with cocaine.
Mice that lack D3 receptors show increased loco-
motor activity and rearing behavior 46,47. A similar
increase in spontaneous locomotor activity was
observed in rats depleted of D3 receptors by injection
of antisense oligonucleotides 4a. Performance of the
D3-receptor null mice in open-field and elevated-plus
maze are characteristic of a reduced level of anxiety.
The D3 receptor might also modify the response to
activation of other DA receptors. These mice also show
an increased sensitivity to cocaine in stimulated loco-
motor activity and to amphetamine in the conditioned-
cue preference test 47. Locomotor stimulation induced
by co-administration of D1- and D2-receptor agonists
is augmented in D3-receptor null mice, but there are
no differences in electrophysiological responses. Genetic
lesioning of both D2 and D3 receptors leads to a more
severe motor phenotype than does D2-receptor lesion
U
Fig. 2. Dopamine in the bindlng-site crevice of the dopamine D2 receptor
viewed from the extracellular side. Dopamine (green) is shown binding
between helices 3, 5 and 6. Sites of interaction are shown in yellow. The
residues are identified according to a consensus numbering scheme
described in Ref. 28. Adapted from Ref. 29, courtesy of J.A. Ballesteros
and J.A. Javitch.
alone 49. D3-receptor null mice have comparable auto-
receptor activity to wild-type mice, indicating that the
D3 receptor does not contribute to autoreceptor func-
tion. Intraventricular injections of antisense oligonucleo-
tides in rats leads to reduced D3-receptor binding,
reduced neurotensin and dynorphin mRNA levels, and
decreased cingulate cortex Fos mRNA levels s°. These
results suggest that D3 receptors tonically modulate
neuropeptide and immediate-early gene expression.
The locomotor effects of the D4 receptor appear to
involve modulation of the responses to activation of
other DA receptors. D4-receptor null mice show
increased sensitivity to cocaine, metamphetamine and
ethanol in locomotor tests sl. The D4 receptor might
also contribute to autoreceptor function, as D4-receptor
deficient mice have increased DA synthesis. A DS-
receptor null mouse line has been developed but not
yet fully characterized 3. Injection of a DS-receptor anti-
sense oligonucleotide potentiates the rotatory response
to SKF38393 in rats with unilateral lesions induced by
6-hydroxydopamine. This contrasts with Dl-receptor
antisense oligonucleotides, which prevent this response.
These results suggest that DS-receptor activation inhibits
locomotor behavior.
Other components of the dopaminergic system
have also been targeted for genetic study. Inactivating
the gene for tyrosine hydroxylase produces dopamine-
depleted mice that are profoundly hypoactive. Mice
deficient in the gene for the DA transporter show
marked hyperactivity and insensitivity to the locomotor
effects of both amphetamine and cocaine. Interestingly,
these mice still self-administer cocaine and the rewarding
effects of morphine are augmented.
The genetic approaches to the study of the function
of the dopaminergic system reveal a complex role for
the various DA receptors in the modulation of loco-
motion. All subtypes studied by gene lesion or over-
expression influence locomotor activity, either directly
or via their modulation of other DA-receptor systems.
In evaluating these models, it has become clear that
current drugs might not differentiate between the
various molecular DA-receptor subtypes in vivo. A
recent study underscores this problem. Agonists and
antagonists that were believed to be D3-receptor selec-
tive were found to have identical effects in wild-type
and D3-receptor null mice 52, More-specific receptor
agonists are required.
'IJNS Vol. 23, No. I0 (Suppl.), 2000 $37
 S.C. Sealfon and C.W. Olanow - Dopamine receptors: f r o m structure t o behavior
Dopamine
Receptor subtypes
Channels , , ~ ~ ~
Intracellular
signaling
network
Cellular
responses
Neuronal
circuits
Behavior
Basal ganglia, Parkinson's disease and levodopa therapy: TINS supplement
Fig. 3. Emergent properties of self-organizing network systems. The
physiological response that emerges at each higher order of the system
results from overall network activity and might not be fully explained by
the activity of any specific individual components.
Possible role of D A receptors in dyskinesia and
neuroprotection in PD
Dyskinesias complicate L-dopa treatment in the
majority of individuals with PD. Recent studies suggest
that dyskinesias result from abnormal pulsatile stimu-
lation of DA receptors, which leads to dysregulation of
downstream genes and proteins, and a consequent
alteration in the firing pattern of basal-ganglia output
neurons s3. However, the precise molecular mechanism
by which pulsatile stimulation of striatal dopamine
receptors is translated into altered firing patterns in
pallidal neurons has not yet been established. Some
molecular changes associated with DA-receptor stimu-
lation and denervation, as well as with the development
of dyskinesia, are only now beginning to be revealed.
DA-receptor stimulation is associated with upregulation
of the expression of immediate-early genes s4. These
gene changes can influence m a n y aspects of neuronal
function including regulation of neurotransmitters and
ion channels, and thus might be crucial for long-term
adaptive responses. The pattern of change in genes and
G proteins is markedly altered by dopaminergic-neuron
S38 TINS VoL 23, No. 10 (Suppl,), 2000
denervation. In the 6-hydroxydopamine-lesioned rat,
there are increased levels of the e~ subunits of G s, Go~f
and G i in the striatum ss'56. Dopaminergic-neuron de-
nervation is also associated with increased levels of
preproenkephalin (PPE) mRNA, and decreased levels
of preprotachykinin in D2- and Dl-receptor-bearing
striatal neurons, respectivelysT'sS.These changes tend to
be reversed with therapy that involves DA-like agents.
However, increased PPE synthesis is not reversed by
L-dopa or short-acting DA-receptor agonists that are
associated with dyskinesia, but is reversed by long-acting
DA-receptor agonists that do not induce dyskinesia.
Indeed, several studies have demonstrated that persis-
tent upregulation of PPE synthesis correlates with the
development of dyskinesia in 1-methyl-4-phenyl-l,2,3,6-
tetrahydropyridine (MPTP) monkey models 5941. These
findings illustrate the potential of pulsatile stimulation
of striatal DA receptors with short-acting dopamine-
like agents to induce changes in gene expression that
might contribute to the induction of altered neuronal
circuits and abnormal behaviors such as dyskinesia.
DA-receptor activation might also protect against
neurodegeneration in PD. Both L-dopa and DA are oxi-
dized to yield reactive oxygen species and can be toxic
to cultured DA neurons 62. However, L-dopa does not
induce degeneration of dopaminergic neurons in normal
rodents, n o n h u m a n primates or humans, suggesting
that, in vivo, protective systems might obviate oxidative
damage. This concern has, however, led to interest in the
potential of DA-receptor agonists, which do not undergo
oxidative metabolism, for providing neuroprotective
effects in PD through a variety of antioxidant mecha-
nisms 62. Recent evidence now suggests that DA-receptor
agonists might also protect dopaminergic neurons
through a receptor-mediated mechanism. For example,
bromocriptine protection of mesencephalic neurons
from L-dopa toxicity is blocked by the D2-receptor
antagonist sulpiride and by antisense oligonucleotides
directed against D2 receptor¢ 3.
Other work has used microarrays to begin to evalu-
ate the genes that are regulated by stimulation of the
DA receptor 64. Stimulation of DA receptors with the
DA-receptor agonist pramipexole induces increased
expression of multiple genes involved in synaptic
plasticity and neuroprotection. It is likely that m a n y
more genes are affected by stimulation of DA receptors,
and that different patterns of gene expression will result
from pulsatile versus continuous stimulation of the recep-
tor and with the use of different DA-like agents. This
pattern might influence the likelihood that dyskinetic
behavior will develop or that a nerve cell will degenerate.
A complexity perspective
Many elements determine the physiological effects
following activation of DA receptors in the normal and
parkinsonian states. Important determinants include
the molecular identity of the receptor stimulated and
the specific signal-transduction pathways that are acti-
vated. A variety of G-protein, ion-channel and second-
messenger systems are modulated following receptor
stimulation 2. The specific intracellular signaling pattern
that results is dependent on the initial state of the cell,
interaction between receptor hetero-multimers and sig-
naling intermediaries, and the precise pattern of receptor
stimulation. Receptor stimulation can induce both
immediate and long-term changes in cell physiology,
and these in turn can be translated into alterations in
 S.C. Sealfon and C.W. O l a n o w - Dopamine receptors: from structure to behavior
Box I. Discussion
Gerfen: I t h i n k it is fascinating to consider t h a t the receptor can exist
in activated a n d unactivated states a n d these are linked to different
G proteins a n d signaling pathways,
Sealfon: We k n o w t h a t these receptor proteins exist in multiple
states. For example, you can define multiple conformations of the
protein, thodopsin, some of w h i c h are active and some n o t active
These conformations generate a different affinity for different G pro-
reins. A specific agonist m i g h t have a higher affinity for one of the
active conformations t h a n anotheI, so it wilt stabilize the receptor in
that particular conformation and cause preferential activation of a
specific signaling pathway. In this way, different agonists m a y
induce different behavioral effects,
Gerfem You have examples of different signaling pathways with
serotonin receptor subtypes. W h a t a b o u t d o p a m i n e receptors?
Sealfon: I d o n ' t k n o w t h a t it's b e e n looked at in t h e d o p a m i n e
receptor as yet, b u t I suspect t h a t it will be t h e same story as w i t h
the serotonin receptor.
Smith: We recently looked at metabotmpic glutamate receptors a n d
found that they were associated with glutamatergic synapses, b u t also
with other types of synapses t h a t have n o t h i n g to do with glutamate,
While they require glutamate activation, it's very likeIy t h a t once
this occurs, these receptor effects will interact w i t h o t h e r types of
receptor effects at these synapses At t h e last Neuroscience meeting
(Miami, 1999) t h e r e was a poster showing t h a t in transfected cells
there is a n interaction between D5 receptors a n d GABAa receptors. If
you transfect celts w i t h D5 receptors and GABAA, the activation of
the D5 receptors plays a role in the internalization of GABAA recep-
tors. The m e c h a n i s m of course is unclear, b u t I t h i n k t h a t it is rea-
sonable to consider that they interact t h r o u g h effects o n G proteins
t h a t mediate specific functions,
Obeso: W o u l d y o u speculate o n w h i c h factors account for a g i v e n
drug acting at t h e receptor for a shorter or longer period of time?
W h a t is the molecular or atomical basis for a molecule w h i c h binds
to a receptor b e c o m i n g inactive at a given m o m e n t ?
Sealfon: The kinetics a n d distribufi°n of a given drug influence its
capacity to b i n d t o a receptor Once you have binding, the specifics of
the atomic interaction influence t h e on- and off-rates of the drug. We
d o n ' t have specific answers as to w h y one drug m i g h t h a v e a longer
action t h a n a n o t h e r ligand, but this could relate to t h e molecular
structure of t h e interaction with t h e receptor a n d the G proteins that
are regulated by t h e interaction between t h e receptor a n d t h e drug.
Walters: If you record from d o p a m i n e n e u r o n s i n t h e rat a n d give
a p o m o r p h i n e , there are short-term reductions in firing rate.
S o m e t h i n g h a p p e n s to t h e receptor though, because if you intro-
duce a n o t h e r dose of apomorphine, you d o n ' t get a response If you
try the same experiment with a m p h e t a m i n e , t h e receptors are m u c h
less likely to b e c o m e desensitized. So w h a t is going on? Do
d o p a m i n e o r d o p a m l n e a g o n i s t s differentiafiy affect receptors?
Olanow: Another question is whether there are molecular or confor-
mational changes in a receptor w h e n it's denervated?
Sealfon: The question you're asking is really a signaling question i n
disguise W h a t happens with desensitization of a receptor is a function
n e u r o n a l survival, n e u r o n a l circuit activity a n d b e h a v i o r .
Understanding the relationship between the pattern
of r e c e p t o r s t i m u l a t i o n , t h e m y r i a d of s i g n a l i n g c o m -
p o n e n t s a n d t h e p h y s i o l o g i c a l effects t h a t e n s u e is dif-
ficult. It is b e c o m i n g i n c r e a s i n g l y a p p r e c i a t e d t h a t t h i s
s y s t e m f o r m s a c o m p l e x n e t w o r k t h a t is c h a r a c t e r i z e d ,
b o t h a t t h e m o l e c u l a r a n d n e u r o n a l level, b y p a t t e r n s
t h a t r e p r e s e n t m o r e t h a n t h e s u m of t h e i n d i v i d u a l
c o m p o n e n t s (Fig. 3). S i m p l e s i g n a l i n g n e t w o r k s i n
invertebrates have demonstrated that multiple trans-
mitters can induce physiological responses, which
c a n n o t b e o b t a i n e d b y e a c h t r a n s m i t t e r a c t i n g a l o n e 6s.
C o m p u t e r s i m u l a t i o n of i n t r a c e l l u l a r s i g n a l i n g n e t w o r k s
REV~gW
of the downstream response. So the question of why one drug
causes a different pattern of receptor desensitization t h a n another is
essentially the signal coding t h a t occurs after receptor stimulation.
Depending o n t h e c o n f o r m a t i o n of t h e receptor t h a t is induced by
the agonist, there may be different types of signal intermediaries
a n d behavioral responses. And essentially, it's t h e same question as
to w h y different ways of administering the same drug m i g h t have
different signaling a n d behavioral effects. This is n o t easy to sort
out, because at t h e molecular level, a given agonist m i g h t activate
three or four different parallel pathways t h a t intersect with each
other. And, predicting h o w this i n f o r m a t i o n is translated into
behavioral responses for different drugs, with different kinetics is
going t o be very, very difficult.
O l a n o w : It seems t h e n that at an intracellular level, you are seeing
t h e interaction of m a n y different molecules t h a t ultimately deter-
m i n e functional effects. It is sort of like an intracellular network.
Sealfon: Exactly. W h e n there was just a D1 a n d a D2 receptor, life
was simple, a n d you cOUld predict a particular effect. As we came to
realize t h a t there are a n u m b e r of different receptors t h a t interact at
m a n y levels, it is clear that any effect we see must relate to all of the
receptors t h a t are activated, t h e G proteins t h a t are regulated, and
t h e effects o n calcium channels t h a t m i g h t cause a phosphorylation
event to occur w i t h inactivation of t h e receptor. We can measure
the effects of a drug o n three or four pathways w h e n we activate a
cell in a very restricted transfected celt system. But, even there our
ability to predict t h e o u t c o m e of h o w receptor activation alters cell
physiology is limited. W h a t we need to do is study the system more
generically using techniques ~ke gene microarrays t h a t allow us to
look at m a n y different genes at the same time. In this way we can
b e g i n to u n d e r s t a n d t h e pattern of change t h a t induces a specific
functional effect. At present, it is difficult if n o t impossible to figure
o u t w h a t behavior will emerge following receptor stimulation w h e n
we really only have the capacity to took at one p o i n t at a time.
Obeso: From a practical p o i n t of view, how will you m a n a g e all of
this data, a n d more specifically, h o w will you k n o w w h i c h signals
are i m p o r t a n t for a particular behavioral response a n d w h i c h ones
are irrelevant.
Sealfon: That is a very i m p o r t a n t question. We know from knock-
o u t models t h a t most molecules are n o t critical because most
knockouts d o n ' t have m u c h of a phenotype. Indeed, the p h e n o -
types are amazingly subtle considering t h a t there is the complete
loss of a p r o t e i n ti~at seemed to b e essential. In m y mind, it's n o t
t h a t the proteins a r e n ' t essential, it's t h a t t h e y a r e non-essential by
themselves a n d t h a t t h e y f u n c t i o n as part of a system. If you get
rid of one little piece of the system~ it's n o t necessarily essential. I
t h i n k it will b e necessary to utilize c o m p u t a t i o n a l approaches a n d
m a t h e m a t i c a l models to u n d e r s t a n d h o w receptor stimulation
rearranges t h e intracellular network leading to a behavioral
response. Ultimately, I believe we will have to u n d e r s t a n d biology
i n m a t h e m a t i c a l terms. We d o n ' t need to k n o w every signal, b u t to
get started we do need a n accurate series of observations to make
these analyses.
s u g g e s t s t h a t d i s c r e t e n e t w o r k s t a t e s c a n e x i s t t h a t are
only loosely tied to any particular signaling com-
p o n e n t 66. N e u r o s c i e n c e is s u c c e e d i n g i n e l u c i d a t i n g t h e
m o l e c u l a r t a r g e t s of D A a n d D A - r e c e p t o r a g o n i s t s , a n d
in mapping each receptor to specific cellular and
b e h a v i o r a l r e s p o n s e s . T h e d e v e l o p m e n t of g e n o m e - w i d e
profiling techniques holds the promise of even better
sampling of the consequences of receptor stimulation.
T h e c h a l l e n g e is t o d e f i n e t h e c o m p l e x r e l a t i o n s h i p s
b e t w e e n r e c e p t o r a c t i v a t i o n , cell s f g n a l i n g , circuit activ-
ity a n d p h y s i o l o g i c a l effects i n q u a n t i t a t i v e t e r m s .
For f u r t h e r d i s c u s s i o n o n t h i s t o p i c see B o x 1.
TINS VoL 23, No. 10 (Suppl.), 2000 $39
 R~-VIEW S.C. Sealfon and C . W . Olanow ~ Dopamine receptors: f r o m structure t o behavior
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