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Striatal enlargement has been consistently reported in schizophrenics receiving chronic neuroleptic treatment although the results following atypical antipsychotic treatment have been equivocal. In order to disentangle patient illness from a possible drug effect on brain structure, young adult rats were administered either haloperidol, risperidone, clozapine, olanzapine, or vehicle daily for four or eight months via drinking water. Significant increases in caudate-putamen volumes were seen in animals receiving either haloperidol or clozapine when compared with control animals following eight months of drug administration. Conversely, olanzapine-treated animals showed significant decreases in caudate-putamen volumes when compared with control animals after eight months of drug. Thus, converging evidence indicates that the neuroplastic response of the striatum following neuroleptic exposure causes volumetric increases, whereas atypical antipsychotics affect the basal ganglia differentially. The current data suggests that such differential responses may be due to both the pharmacological properties and the relative doses of the atypical agents.

D(2)-like dopamine receptors mediate functional changes via activation of inhibitory G proteins, including those that affect adenylate cyclase activity, and potassium and calcium channels. Although it is assumed that the binding of a drug to a single isoform of a D(2)-like receptor will cause similar changes in all receptor-mediated functions, it has been demonstrated in brain that the dopamine agonists dihydrexidine (DHX) and N-n-propyl-DHX are "functionally selective". The current study explores the underlying mechanism using transfected MN9D cells and D(2)-producing anterior pituitary lactotrophs. Both dopamine and DHX inhibited adenylate cyclase activity in a concentration-dependent manner in both systems, effects blocked by D(2), but not D(1), antagonists. In the MN9D cells, quinpirole and R-(-)-N-propylnorapomorphine (NPA) also inhibited the K(+)-stimulated release of [(3)H]dopamine in a concentration-responsive, antagonist-reversible manner. Conversely, neither DHX, nor its analogs, inhibited K(+)-stimulated [(3)H]dopamine release, although they antagonized the effects of quinpirole. S-(+)-NPA actually had the reverse functional selectivity profile from DHX (i.e., it was a full agonist at D(2L) receptors coupled to inhibition of dopamine release, but a weak partial agonist at D(2L) receptor-mediated inhibition of adenylate cyclase). In lactotrophs, DHX had little intrinsic activity at D(2) receptors coupled to G protein-coupled inwardly rectifying potassium channels, and actually antagonized the effects of dopamine at these D(2) receptors. Together, these findings provide compelling evidence for agonist-induced functional selectivity with the D(2L) receptor. Although the underlying molecular mechanism is controversial (e.g., "conformational induction" versus "drug-active state selection"), such data are irreconcilable with the widely held view that drugs have "intrinsic efficacy".

Dihydrexidine (DHX), the first high-affinity D 1 dopamine receptor full agonist, is only 10-fold selective for D 1 versus D 2 receptors, having D 2 affinity similar to the prototypical agonist quinpirole. The D 2 functional properties of DHX and its more D 2 selective analog N - n -propyl-dihydrexidine (PrDHX) were explored in rat brain and pituitary. DHX and PrDHX had binding characteristics to D 2 receptors in rat striatum typical of D 2 agonists, binding to both high- and low-affinity sites and being sensitive to guanine-nucleotides. Consistent with these binding data, both DHX and PrDHX inhibited forskolin-stimulated cAMP synthesis in striatum with a potency and intrinsic activity equivalent to that of quinpirole. Unexpectedly, however, DHX and PrDHX had little functional effect at D 2 receptors expressed on dopaminergic neurons that mediate inhibition of cell firing, dopamine release, or dopamine synthesis. Quantitative receptor competition autoradiography demonstrated that DHX bound to D 2 receptors in striatum (predominantly postsynaptic receptor sites) with equal affinity as D 2 sites in the substantia nigra (autoreceptor sites). The data from these experiments, coupled with what is known about the location of specific dopamine receptor isoforms, lead to the hypothesis that DHX, after binding to D 2L and D 2S receptors, causes agonist-typical functional changes only at some of these receptors. This phenomenon (herein termed “functional selectivity”) suggests that drugs may be targeted not only at specific receptor isoforms but also at separate functions mediated by a single isoform, yielding novel approaches to drug discovery.

The unavailability of selective D1A (D1) or D1B (D5) dopamine receptor ligands has prevented the direct localization of binding sites for these receptors. Thus, receptor autoradiography with long exposure times was used to detect minor D1-like binding sites in the brains of D1A null mutants. Coronal brain sections were prepared from the caudal portion of the prefrontal cortex of homozygous or heterozygous D1A knockout mice or wildtype mice, and labeled with the D1 receptor antagonist [3H]-SCH23390. Slides were dried, and apposed to film with polymer-calibrated standards for 90 days to allow visualization of any low abundance binding sites. No binding was detected in most regions of homozygote (−/−) mouse brains that have high densities of D1 binding in wildtype mice (e.g., the striatum, nucleus accumbens, olfactory tubercles or amygdala). Conversely, small, but detectable amounts of D1-binding were measured in the hippocampus, albeit with a density less than the lowest standard (ca. 20 fmol/mg). Saturation binding of [3H]-SCH23390 in hippocampal homogenates from homozygous mice confirmed a Bmax of 12.3 fmol/mg protein with a KD of 0.57 nM. The current work demonstrates directly the presence of D1B (D5) receptors in hippocampus, and also shows that the loss of functional D1A gene products almost completely eliminates detectable D1-binding sites in striatum, as well as in some regions (e.g., the amygdala) where a non-adenylyl cyclase coupled D1 receptor has been reported. This indicates that these non-adenylyl cyclase coupled D1-like receptors represent alternate signaling pathways rather than novel gene products(s). Synapse 39:319–322, 2001. © 2001 Wiley-Liss, Inc.

Dopamine D 1A receptor-deficient mice were assessed in a wide variety of tasks chosen to reflect the diverse roles of this receptor subtype in behavioural regulation. The protocol included examination of exploration and locomotor activity in an open field, a test of sensorimotor orienting, both place and cue learning in the Morris water maze, and assessment of simple associative learning in an olfactory discrimination task. Homozygous mice showed broad-based impairments that were characterized by deficiencies in initiating movement and/or reactivity to external stimuli. Data obtained from flash evoked potentials indicated that these deficits did not reflect gross visual impairments. The partial reduction in D 1A receptors in the heterozygous mice did not affect performance in most tasks, although circumscribed deficits in some tasks were observed (e.g., failure to develop a reliable spatial bias in the water maze). These findings extend previous behavioural studies of null mutant mice lacking D 1A receptors and provide additional support for the idea that the D 1A receptor participates in a wide variety of behavioural functions. The selective impairments of heterozygous mice in a spatial learning task suggest that the hippocampal/cortical dopaminergic system may be uniquely vulnerable to the partial loss of the D 1A receptor.

A critical assumption in the unilateral 6-hydroxydopamine (6-OHDA) model is that interactions between the intact and denervated hemispheres do not influence the response to insult. The present study examined this issue by assessing the effects of unilateral substantia nigra 6-OHDA lesions in rats that previously had received corpus callosum transections, a treatment designed to minimize interhemispheric influences. Quantitative autoradiography in the caudate-putamen ipsilateral to the lesion revealed that corpus callosum transection did not alter the increase in D2-like receptors ([125I]-epidepride-labeled sites) that is induced by unilateral 6-OHDA lesion. There were no effects of either 6-OHDA lesion or transection on D1 receptor density ([l25I]-SCH23982 autoradiography). As a functional endpoint, dopamine-stimulated cAMP efflux was measured in superfused striatal slices. In this paradigm, the net effect of dopamine (DA) represents a combination of D1 receptor-mediated stimulation and D2 receptor-mediated inhibition. 6-OHDA lesion increased cAMP efflux induced by exposure to 100 p.M DA alone; corpus callosum transection did not alter this effect. An interaction between 6-OHDA lesion and transection status was revealed, however, by comparison of results obtained with DA alone vs. DA plus the D2 antagonist sulpiride (to block the D2 inhibitory effects of 100 p.M DA). This comparison revealed two important effects of 6-OHDA lesion in rats with an intact corpus callosum: (1)a moderate decrease in dopamine Dl receptor-mediated stimulation; and (2) a dramatic decrease in the ability of D2 receptors to inhibit this stimulation. Corpus callosum transection prevented these effects of 6-OHDA. These results provide a biochemical demonstration of D1:D2 receptor uncoupling in unilateral 6-OHDA lesioned rats, and suggest that interhemispheric influences (e.g., contralateral cortico-striatal glutamatergic projections) may contribute to lesion-induced alterations in D1:D2 receptor interactions. © 1995 Wiley-Liss, Inc.

The effects of postmortem interval on dopamine D1 mRNA and D1 receptors were assessed in rat striatum under conditions simulating the handling of human brain tissue at 0, 6, 12, and 24 h postmortem. The amount of D1 mRNA was measured by both in situ hybridization film and emulsion autoradiography with [35S]dATP-labeled oligonucleotide probes. D1 receptor density was determined by autoradiography with [125I]SCH 23982. Neither the total amount of D1 mRNA in the striatum nor the frequency distribution of striatal cells expressing D1 mRNA varied with the postmortem interval. There was a modest but significant decrease (ca. 10%) in D1 receptors over the 24 h postmortem interval; this decrease occurred within the first 6 h postmortem, with no further decreases up to 24 h postmortem. These findings suggest that the effects of postmortem interval on D1 mRNA and receptors are minimal and should not limit an examination of possible alterations in dopamine D1 receptor mRNA and D1 receptors in the postmortem brains of humans with neuropsychiatric disease.

In an analogy to the potent catechol dopamine D1 agonists dihydrexidine (1) and dinapsoline (2), benzo rings were fused onto the structures of the dopamine D2-selective agonists quinelorane (3) and quinpirole (4). Each of the phenyl ring-substituted derivatives had significant affinity for D2 receptors, albeit somewhat lower than the two parent compounds, 3 and 4. Compounds with N-propyl and N-allyl substituents (5b, 5c, 6c, and 6d) had higher affinity for the D2 dopamine receptor than did their corresponding secondary amines (5a and 6a). Slightly different effects on affinity of an n-propyl and an n-allyl group in the new analogues of 3 and 4 suggest that different binding orientations may be invoked at the receptor.

The N-n-propyl analog of dihydrexidine ((±)-trans-10,11-dihydroxy-5,6,6a,7,8,12b-hexahydrobenzo[a]phenanthridine) is a dopamine receptor agonist with high affinity for dopamine D2 and D3 receptors (K0.5=26 and 5 nM, respectively). Members of the hexahydrobenzo[a]phenanthridine structural class are atypical because they display high intrinsic activity at post-synaptic dopamine D2 receptors, but low intrinsic activity at dopamine D2 autoreceptors. The present study examined the effects of (±)-N-n-propyl-dihydrexidine on unconditioned behaviors in rats. The most striking results observed were large, dose-dependent decreases in locomotor activity (e.g., locomotor inhibition), and increases in vacuous chewing; yawning was also increased at the highest dose of (±)-N-n-propyl-dihydrexidine. The locomotor inhibition and yawning induced by (±)-N-n-propyl-dihydrexidine were blocked by pre-treatment with (−)-remoxipride (S(−)-3-bromo-N-((1-ethyl-2-pyrrolidinyl)-methyl)-2,6-dimethoxybenzamide), a dopamine D2 receptor antagonist, but not by the dopamine D1 receptor antagonist (+)-SCH23390 (R(+)-7-chloro-8-hydroxy-3-methyl-1-phenyl-2,3,4,5-tetrahydro-1H-3-benzazepine). Vacuous chewing was decreased by both (−)-remoxipride and (+)-SCH23390. These data support the hypothesis that a subpopulation of post-synaptic dopamine D2 receptors has a critical role in decreases in locomotor activity and induction of vacuous chewing and yawning. © 1997 Elsevier Science B.V. All rights reserved.

The intrinsic activities of selected dopamine D1 receptor agonists were compared in three distinct molecular expression systems, C-6, Ltk, and GH4, cells transfected with primate D1A receptors. The influence of the cell expression system on intrinsic activity varied markedly among agonists. Dihydrexidine (DHX), a potent full agonist with dramatic antiparkinsonian actions, displayed intrinsic activity similar to dopamine in all three cell lines. In contrast, SKF82958 and SKF38393 (full and partial agonists, respectively, in rat striatum) had intrinsic activities equal to dopamine in GH4, cells that expressed a high density of D1 receptors, yet were of lower intrinsic activity in C-6 cells having 15-fold fewer receptors. The idea that spare receptors are one important determinant of observed intrinsic activity was explored directly by “receptor titration”, in which ca. 90% of D1 receptors in Ltk cells were inactivated using EEDQ, an irreversible antagonist. Whereas EEDQ pretreatment decreased the potency of all agonists, it changed the intrinsic activity of some, but not all, drugs. A 40% decrease was seen with the partial agonist SKF38393, and, surprisingly, a 30% decrease was seen with the purported full agonist SKF82958. Conversely, the intrinsic activity of DHX and A68930 were unaffected by the EEDQ treatment. The data demonstrate that significant and biologically meaningful differences in intrinsic efficacy (e.g., DHX vs. SKF82958) may be obscured in test systems that have sufficient receptor reserve (e.g., the striatum). Such differences in intrinsic efficacy may be an important predictor of the clinical utility of D1 agonists. © 1995 Wiley-Liss, Inc.

Recent data indicate that full D1 dopamine agonists have greater antiparkinsonian effects in the MPTP primate model than do partial agonists, suggesting that the intrinsic activity of D1 agonists may affect their utility in the treatment of Parkinson's disease. It is unclear, however, whether human D1 receptors in situ are similar to D1 receptors in other species or in molecular expression systems. For this reason, the binding affinity and functional activity of a series of D1 dopamine receptor agonists [dihydrexidine (DHX), SKF82958, and A68930] were determined in postmortem human caudate. Results from in vitro binding studies with membranes from human caudate indicate that these D1 agonists competed for [3H]SCH23390 labeled sites with a rank order similar to that found in rat striatum [K50 = 36.8 nM (DHX); 18.6 nM (SKF82958); 3.9 nM (A68930)]. The ability of these compounds and the partial agonist SKF38393 to stimulate the enzyme adenylyl cyclase in tissue homogenates of human caudate was also examined. DHX and A68930 are full agonists compared to dopamine, whereas SKF82958 and SKF38393 are partial agonists. These differences in biochemical intrinsic activity are consistent with the profound antiparkinsonian effects caused by DHX, but not by SKF82958 and SKF38393, in the MPTP-monkey model. This suggests that DHX and A68930 may be of greater utility in treating disorders where a full efficacy D1 agonist may be required.

Changes in the mesostriatal dopamine system associated with normal aging are observed in both human and laboratory animals, but the specific behavioral consequences of these nonpathological changes are largely unexplored. The present study (a) assessed the effects of normal aging on markers for the mesostriatal dopamine system, and (b) examined the relationship of age-related changes in this system to decline in reaction time performance. Decreased levels of midbrain dopamine (DA) and dihydroxyphenylacetic acid (DOPAC) were observed in the aged rats as compared to young, but there was no evidence for age-related changes in the density of D1 or D2 receptor binding or the density of dopamine uptake sites. Some differences were observed when the aged rats were grouped according to reaction time performance. Aged RT-unimpaired rats exhibited higher density of D1 binding in rostrodorsal striatal patch areas, but lower overall levels of DA. In caudal striatum, aged RT-unimpaired rats exhibited lower DA and higher DOPAC levels.

Although partial efficacy dopamine D1 receptor agonists have little therapeutic benefit in parkinsonism, the first high potency, full efficacy dopamine D1 receptor agonist dihydrexidine recently has been shown to have profound antiparkinsonian effects. One reason for the greater antiparkinsonian effects of dihydrexidine vs. SKF38393 might be that SKF38393, while a partial dopamine D1 receptor agonist in rodent striatal preparations, has virtually no agonist activity in monkey striatum (Pifl et al., 1991, Eur. J. Pharmacol. 202, 273). To explore this hypothesis, we compared the dopamine D1 receptor affinity and efficacy of dihydrexidine and SKF38393 in striatum from rat and monkey. In vitro binding studies using membranes from putamen of adult rhesus monkeys demonstrated that dihydrexidine competed for dopamine D1 receptors (labeled with [3H]SCH23390) with high potency (IC50 = 20 nM vs. ca. 10 nM in rat brain). SKF38393 was about 4-fold less potent than dihydrexidine in both monkey and rat brain. The in vitro functional activity of these drugs was assessed by their ability to stimulate adenylate cyclase activity in tissue homogenates. Dihydrexidine was of full efficacy (relative to dopamine) in stimulating cAMP synthesis in both monkey and rat. SKF38393 was only a partial efficacy agonist in both rat striatum and monkey putamen, but contrary to the original hypothesis, it had the same efficacy (ca. 40% relative to dihydrexidine) in membranes from both species. Interestingly, greater between-subject variation was found in the stimulation produced by SKF38393 in primate compared to rat brain, although the basis for this variation is unclear.(ABSTRACT TRUNCATED AT 250 WORDS)

We report that certain substituted hexahydrobenzo[a]phenanthridines are novel high affinity ligands selective for the dopamine D3 receptor. These data demonstrate that substitutions on the heterocyclic nitrogen and the pendant phenyl ring of this nucleus cause a marked increase in both affinity and selectivity for dopamine D3 vs. D2 receptors. Thus, these compounds represent important new tools to study the pharmacology of dopamine D3 receptors, and may also provide an opportunity for the synthesis of new radioligands for dopamine D3 receptors.