D-1/D-2 behavioural interactions in the rat involving striatal dopamine D-1 receptors

Simultaneous systemic administration of SKF 38393 (1-15 mg/kg i.p.) and LY 171555 (0.625-20 mg/kg s.c.) showed clear evidence of dopamine D-1/D-2 behavioural interactions compared to either treatment given alone. Similar interactions were observed between an intermediate systemic dose of LY 171555 (5 mg/kg) and SKF 38393 (1-10 micrograms) microinjected bilaterally into the caudate-putamen, but not into the substantia nigra pars reticulata, suggesting that striatal dopamine D-1 receptors are the ones responsible for mediating the altered behavioural responses to D-2 agonists in the intact rat.     

Aporphine enantiomers. Interactions with D-1 and D-2 dopamine receptors

The R(-) and the S(+) enantiomers of apomorphine (APO) and N-n-propyl norapomorphine (NPA) interact with both the D-1 and the D-2 dopamine receptors. R(-)-APO, as well as R(-)- and S(+)-NPA, stimulates the D-1 dopamine receptor in carp retina; S(+)-APO blocks this dopamine receptor. Similarly, R(-)-APO, as well as R(-)- and S(+)-NPA, stimulates the D-2 dopamine receptor in the intermediate lobe of the rat pituitary gland; S(+)-APO blocks the intermediate lobe D-2 receptor. The interactions between these aporphine enantiomers and the D-1 and the D-2 dopamine receptors exemplify several manifestations of the previously described "n-propyl phenomenon." Because S(+)-APO is distinguished from the other tested aporphines by its ability to antagonize either the D-1 or the D-2 dopamine receptors, it is hypothesized that the presence of an N-methylated tertiary amine in a molecule of appropriate configuration can confer dopamine receptor antagonist activity to the molecule.     

SCH 23390 blocks D-1 and D-2 dopamine receptors in rat neostriatum in vitro

Summary The agonistic and antagonistic effects of the new compound SCH 23390 were tested in functional model systems for the D-1 dopamine receptor and for the D-2 dopamine receptor in vitro. In superfused rat neostriatal slices the increase in the efflux of cyclic AMP was used as a parameter for D-1 receptor stimulation. D-2 receptor stimulation was measured as the decrease in the K⁺-evoked release of [³H]-acetylcholine. SCH 23390 had no agonistic activity in these two models. SCH 23390 was a potent antagonist of the stimulating effect of dopamine in the D-1 receptor model (apparent pA₂=7.28). SCH 23390 also antagonized the effect of the D-2 receptor agonist LY 141865 in the D-2 receptor model (apparent pA₂=6.34). This D-2 receptor antagonism proved to be of a competitive nature.     

Stimulation of D-1 dopamine receptors facilitates D-2 dopamine receptor recovery after irreversible receptor blockade

The hypothesis that stimulation of the D-1 dopamine receptor subtype affects the recovery of the D-2 subtype after alkylation by EEDQ was investigated. Animals were pretreated with either SCH23390, to protect D-1 receptors, or saline, before administration of EEDQ. After EEDQ one group of saline pretreated animals received 12 hourly injections of the D-1 agonist SKF38393. Animals were sacrificed at 6, 24 and 48 hours after EEDQ and Kd and Bmax of striatal D-1 and D-2 receptors measured. The concentration of D-2 receptors in the groups in which D-1 receptors had been protected by SCH23390 or stimulated by SKF38393 were significantly greater than that of the EEDQ alone group.     

3H]dopamine binds to D-1 and D-2 receptors in rat striatum

Although saturation studies suggest that [3H]dopamine binds to a homogeneous class of stereospecific sites in rat striatal membranes, pharmacologic analysis reveals two distinct sites, one with high and another with low antagonist affinities. The affinity of antagonists for the first site is correlated with their affinity for [3H]butyrophenone binding sites; affinity for the second site is correlated with inhibitory potency against the dopamine-stimulated adenylate cyclase. These results suggest that [3H]dopamine binds to D-1 and D-2 receptors.     

Effects of adrenalectomy on responses mediated by dopamine D-1 and D-2 receptors

The effects of surgical adrenalectomy were investigated on behavioural responses produced by the selective D-1 agonist, SK&F 38393, alone, and in combination with the D-2 agonist, quinpirole (LY171555). Further, stereotyped responses to apomorphine and LY171555 were assessed following treatment with either the D-1 or the D-2 antagonists, SCH 23390 and raclopride, respectively. There was no difference between sham-adrenalectomized (sham) and adrenalectomized (ADX) groups in responses to SK&F 38393. Although concomitant stimulation of both receptor subtypes increased the incidence of stereotyped sniffing behaviour, there was no difference in the magnitude of this effect between the sham and ADX groups. Raclopride reduced LY171555-induced sniffing and hypothermia less in ADX rats than in sham controls, which was consistent with the hypothesis that adrenocortical hormones affect D-2 receptor responsiveness. SCH 23390 had a greater inhibitory effect on LY171555 responses, but a smaller effect on apomorphine responses in the ADX group compared with their sham controls. It is concluded that the amplified D-2-stimulated response observed in ADX rats may be more dependent on tonic D-1 receptor activation than the control D-2 response of shams.     

Inactivation of dopamine D-1 or D-2 receptors differentially inhibits stereotypies induced by dopamine agonists in rats

Ex vivo D-1 or D-2 receptor binding in the striatum was reduced by 65-78% after treatment with EEDQ (N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline) in combination with either the D-2 antagonist, raclopride, or the D-1 antagonist, SCH 23390, respectively. EEDQ induced a 65% reduction in D-1 receptor binding and a 51% decrease in cAMP production in striatal homogenates. Selective D-2 receptor inactivation inhibited the stereotyped behaviour induced by the mixed D-1/D-2 agonist, apomorphine, or by the D-2 agonist, quinpirole, when given alone and in combination with the D-1 agonist, SK&F 38393. Selective inactivation of D-1 receptors did not inhibit the behavioural effects of quinpirole when given alone and in combination with the D-1 agonists, SK&F 81297, SK&F 38393 or SK&F 75670. Likewise, the effect of apomorphine was unchanged. These results indicate that a normal density of D-2 receptors is critical for the expression of the stereotyped behaviour induced by DA agonists. In contrast, there is a large surplus of D-1 receptors to enable the response to a D-2 agonist. This is particularly illustrated by the persistent behavioural effects of the partial D-1 agonist, SK&F 75670, in rats with up to a 78% decrease in D-1 receptor binding.     

Potentiation of licking in rats by stimulation of both D-1 and D-2 dopamine receptors

Apomorphine-induced licking in rats was assessed by recording the total number of licks by direct observation. Apomorphine induced licking dose dependently. The maximum response was obtained by 0.5 mg/kg of the drug and 30 min after drug administration. Pre-treatment with dopamine antagonists, sulpiride and SCH 23390 decreased the apomorphine effect. Pre-treatment of animals with reserpine+α-methyl-p-tyrosine (AMPT) increased apomorphine-induced licking. In normal rats the D-2 agonist quinpirole and the D-1 agonist SKF 38393 also induced significant licking. The effect of quinpirole or SKF 38393 was decreased by reserpine+AMPT pre-treatment. Combined treatment with SKF 38393 and quinpirole induced more intense licking in both reserpinized and non-reserpinized animals. It is therefore concluded that the apomorphine-induced licking is mediated through both D-1 and D-2 receptors, and that pre-treatment with reserpine hypersensitizes these receptors to the drug effect. However, for either SKF 38393- or quinpirole-induced licking, the presence of endogenous dopamine seems essential.     

Blockade of both D-1 and D-2 dopamine receptors may induce catalepsy in mice.

1. The catalepsy induced by dopamine antagonists has been tested and the possible dopamine subtypes involved in catalepsy was determined. 2. Dopamine antagonist fluphenazine, D-1 antagonist SCH 23390 or D-2 antagonist sulpiride induced catalepsy. The effect of fluphenazine and sulpiride was dose-dependent. Combination of SCH 23390 with sulpiride did not induce catalepsy potentiation. 3. D-1 agonist SKF 38393 or D-2 agonist quinpirole decreased the catalepsy induced by fluphenazine, SCH 23390 or sulpiride. 4. Combination of SKF 38393 with quinpirole did not cause potentiated inhibitory effect on catalepsy induced by dopamine antagonists. 5. The data may indicate that although D-2 receptor blockade is involved in catalepsy, the D-1 receptor may plan a role.     

Selective blockade of dopamine D-1 receptors by SCH 23390 discloses striatal dopamine D-2 receptors mediating the inhibition of adenylate cyclase in rats

l-Glutamate but not methyl-D-aspartate (NMDA) or quisqualate (Quis) (10^?6 M) in vitro with or without preincubation increased significantly the K_D value of the [³H]N-propylnorapomorphine ([³H]NPA) binding sites by 21 and 36% respectively in striatal membranes of rat without influencing the striatal [³H]spiperone binding sites. The number of striatal [³H]NPA binding sites was not changed by l-glutamate (10^?6 and 10^?5 M) in vitro. There may thus exist interactions between striatal glutamate receptors — not related to excitatory amino-acid receptors of the NMDA or the QUIS type - and high affinity striatal DA receptors.     

Role of dopamine D-1 and D-2 receptors in the ventral striatum in the turning behaviour of rats

Systemically administered methamphetamine and apomorphine evoked characteristic turning when rats were pretreated with injections of D-1 and D-2 antagonists in the ventral but not in the dorsal striatum. The frequency of turning was greater for rats pretreated with D-2 antagonists (l-sulpiride and YM-09151-2) than for rats pretreated with a D-1 antagonist (SCH 23390). The responses were apparently additive when rats were pretreated with mixed D-1/D-2 antagonists, (cis(Z)-flupentixol and SCH 23390 + l-sulpiride). These results suggest that dopamine D-1 and D-2 receptors in the ventral striatum mediate turning behaviour through separate but cooperative mechanisms.     

BHT-920 and LY-171555 (quinpirole) have similar affinities for striatal D-2 dopamine receptors, and similar affinities for striatal D-1 dopamine receptors

The affinity of LY-171555 (quinpirole) and BHT-920 for both states of rat striatal dopamine D-1 and D-2 receptors was determined. Although these drugs have different pharmacological effects in experimental animals, we found that they had similar affinities for both D-1 and D-2 receptors.     

EEDQ, a tool for ex vivo measurement of occupancy of D-1 and D-2 dopamine receptors

EEDQ (N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline) inactivates dopamine (DA) D-1 and D-2 receptors, measured by ex vivo [3H]SCH 23390 and [3H]spiperone binding in striatal homogenates. SCH 23390 (D-1 antagonist) and SK&F 38393 (D-1 agonist) protect against inactivation of D-1 receptors (ED50 values 0.075 and 53 mumol/kg, respectively). Raclopride (D-2 antagonist) and quinpirole (D-2 agonist) protect against inactivation of D-2 receptors (ED50 values 0.48 and 7.1 mumol/kg, respectively). The potencies correspond closely to those obtained by in vivo binding experiments using radioligands.     

Aporphines as antagonists of dopamine D-1 receptors

The aporphine alkaloids are a class of compounds known to possess activity at both D-1 and D-2 dopamine receptors. (R)-Apomorphine and (S)-bulbocapnine are examples of compounds which have agonist and antagonist activity, respectively, at D-1 receptors. A series of optically pure aporphines was synthesized and their activity at D-1 and D-2 dopamine receptors was studied. The (R)-aporphines uniformly had greater affinity for both D-1 and D-2 receptors than their S antipodes. Dihydroxy compound (R)-apomorphine, in accord with previous studies, was found to be a D-1 agonist. Aporphines possessing a single hydroxy group at C-11 are antagonists at the D-1 receptor. The corresponding methoxy compounds are virtually inactive at dopamine receptors. The most potent compounds, (R)-11-hydroxyaporphine (R-14) and (R)-10-bromo-11-hydroxyaporphine (R-26), are more potent than bulbocapnine as D-1 antagonists but are not as selective. A model for binding of aporphines to the D-1 receptor was formulated in which binding interactions between the receptor and the basic nitrogen and the C-11 hydroxy group of the aporphine are required for high-affinity binding to the receptor. The absolute configuration at C-6a determines the orientation of the N-6 lone pair and binding is optimal for the 6aR series. The agonist or antagonist activity of an aporphine is determined by the presence or absence, respectively, of a hydroxy group at C-10. A hydrophobic binding site may be present and may account for the high antagonist activity of (S)-bulbocapnine.     

Comparative study of the EEG profile of neuroleptics selective for D-1 or D-2 dopamine receptors in the rabbit.

The neuroleptics SCH 23390 and raclopride, which interact selectively with either D-1 or D-2 dopamine receptor, were studied for their effects on electroencephalographic (EEG) activity in the rabbit. Haloperidol (0.3 and 1 mg/kg intravenously, i.v.), which was used for comparison, induced synchronization of the cortical EEG activity. Spectral EEG analysis showed increase of power in the whole frequency range (0.1-38.5 Hz) and in all frequency bands in the cortex, whereas a slight decrease of slow and fast theta activity (3.7-7.2 and 7.2-12.2 Hz) was observed in the hippocampus. Animals appeared sedated and arousal response to somatosensory stimuli was markedly inhibited. SCH 23390 (0.03 and 0.3 mg/kg i.v.) induced periods of cortical synchronization and changes of spectral power qualitatively similar to those accompanying haloperidol administration. The drug slightly reduced the duration of arousal elicited by stimuli. Raclopride (1 and 3 mg/kg i.v.) induced weak EEG changes and little effect on arousal response to stimulation. There was an increase of slow wave activity which was particularly evident in the hippocampus. The data indicate that, although to a lesser degree, the D-1 receptor antagonist SCH 23390 induced EEG effects similar to those of haloperidol, whereas blockade of D-2 receptors by raclopride resulted in different patterns of EEG activity.     

Behavioural correlates to the dopamine D-1 and D-2 antagonists

The acute dopamine (DA) receptor blockade of neuroleptics can be demonstrated in mice by antagonism of stereotypies induced by the DA-agonist methylphenidate and in rats by antagonism of stereotypies induced by the DA-agonists amphetamine or apomorphine. Neuroleptics such as the thioxanthene, cis(Z)-flupentixol, the phenothiazine, fluphenazine, the butyrophenone, haloperidol and the benzamide clebopride are equipotent behaviourally as well as clinically. Also the D-1 receptor-antagonist SCH 23390 has the same pharmacological effects. In a series of experiments where the methylphenidate-induced stereotyped gnawing in mice was inhibited by neuroleptics it was shown that the effect of butyrophenones was greatly attenuated by concomitant treatment with scopolamine and diazepam. Similar results were obtained in rats experiments. The effect of phenothiazines was less influenced and that of thioxanthenes and SCH 23390 remained nearly unchanged. Besides, a clear differentiation of these drugs was seen when they were tested in mice rendered supersensitive by 12 days treatment with different neuroleptics. In the withdrawal phase the decrease effects against methylphenidate were shown by increased ED50 values for methylphenidate antagonism and an increased response to methylphenidate. The thioxanthenes and SCH 23390 retained the ability to antagonize the stereotyped gnawing, the phenothiazines showed a reduced effect, whereas the butyrophenones showed both tolerance and cross tolerance to the stereotyped behaviour. This behavioural classification of neuroleptics into three different groups is comparable with the classification obtained by DA-receptor binding techniques in vitro.     

Opposing roles for D-1 and D-2 dopamine receptors in regulating the excitability of growth hormone-producing cells in the snail Lymnaea stagnalis

Dopamine hyperpolarizes growth hormone-producing cells (GHC) in the CNS of Lymnaea stagnalis. This effect of dopamine was mimicked by the D-2 receptor agonist LY 141865 and antagonized by the D-2 receptor antagonists (-)-sulpiride and YM 09151-2. SKF 38393, a selective D-1 receptor agonist, increased the excitability of the GHC. This effect was mimicked by intracellular injection of cyclic AMP and antagonized by the D-1 receptor antagonist SCH 23390. Dopamine (in the presence of (-)-sulpiride) also increased the excitability of the GHC. It is concluded that both a D-1 and a D-2 receptor regulate the electrical activity of the GHC in the CNS of Lymnaea stagnalis.     

Studies on the involvement of dopamine D-1 and D-2 receptors in the anticonvulsant effect of dopamine agonists in various rodent models of epilepsy

Dopamine agonists with different selectivity for dopamine D-1 and D-2 receptors in the brain were tested for their effects: on thresholds for maximal electroshock seizures in mice and rats and for pentylenetetrazol-induced clonic seizures in mice; on seizures induced by air blast stimulation in gerbils, and on seizures induced by amygdala-kindling in rats. The mixed D-1/D-2 agonist apomorphine exerted anticonvulsant effects in all models except kindling. In gerbils and mice, the anticonvulsant action of apomorphine could be antagonized by the D-2 selective dopamine antagonist sulpiride. When injected alone, sulpiride exerted no significant effect on seizure activity. The preferential D-2 receptor agonists lisuride and (+)-PHNO [+)-4-propyl-9-hydroxynaphthoxazine) differed in their profile of action. Both compounds displayed anticonvulsant efficacy in gerbils, while only lisuride proved capable of reducing kindled seizure severity. (+)-PHNO increased the threshold for electroconvulsions in mice while lisuride was ineffective in this respect or even decreased the threshold. The reverse was obtained in regard to electroshock seizures in rats. The threshold for seizures induced by pentylenetetrazol in mice was increased significantly by lisuride but not by (+)-PHNO. The selective dopamine D-1 receptor agonist SKF 38393-A exerted no anticonvulsant effect in any seizure test except a moderate increase of the electroconvulsive threshold in mice. In contrast, the dopamine precursor L-DOPA (injected after pretreatment with carbidopa) proved capable of reducing seizure activity in all models. In mice, the increase in the threshold for maximal electroshock seizures induced by L-DOPA was significantly reduced by sulpiride, which also attenuated the anticonvulsant effect of L-DOPA in gerbils. Collectively, the data indicate that dopamine D-2 receptors mediate the anticonvulsant effect of dopamine agonist and, at least in part, of L-DOPA whereas D-1 receptors seem not to be involved.     

Dopamine D-1 and D-2 receptors in Huntington's disease

Dopamine receptors were studied in post-mortem brains from control and Huntington's disease patients, using the specific binding of [3H]spiperone to dopamine D-2 receptors and [3H]piflutixol to dopamine D-1 receptors. Both [3H]spiperone binding and [3H]piflutixol binding were reduced by 45-50% in Huntington's disease putamen. The loss of [3H]spiperone and [3H]piflutixol binding sites correlated with decreased GABA concentrations observed in Huntington's disease putamen. A selective loss (48%) of [3H]piflutixol binding was observed in Huntington's disease substantia nigra pars reticulata, [3H]piflutixol binding was unchanged in substantia nigra pars compacta. No differences in [3H]spiperone binding were observed between the groups in either region of substantia nigra. The results are discussed in relation to the pathophysiology of Huntington's disease, and to the presence of distinct dopamine receptors in human brain.