Comparison of the reinforcing efficacy of two dopamine D2-like receptor agonists in rhesus monkeys using a progressive-ratio schedule of reinforcement

High-efficacy D2-like dopamine (DA) receptor agonists can function as positive reinforcers when made available to animals for intravenous self-administration under a fixed-ratio (FR) schedule of reinforcement. In a previous study, however, low-efficacy D2-like agonists failed to maintain self-administration under an FR schedule, suggesting that agonist efficacy is directly related to efficacy as a positive reinforcer. To examine this hypothesis further, the present study compared two D2-like DA receptor agonists that maintained FR responding, but differ in their D2-like receptor efficacy and selectivity, using a procedure designed to rank-order drugs according to their efficacy as reinforcers. Rhesus monkeys (n=5) were prepared with chronic, indwelling intravenous catheters and allowed to self-administer cocaine (0.1 mg/kg/injection) or saline on different days under a progressive-ratio (PR) schedule. When responding was stable, doses of the full D2-like agonist R(-)-propylnorapomorphine (NPA) or the partial D2-like agonist R(-)-apomorphine (APO) were made available for self-administration in the test sessions. Both compounds maintained self-administration with sigmoidal or biphasic dose-response functions. Surprisingly, the lower efficacy agonist APO was the more efficacious positive reinforcer. This result fails to support the hypothesis that D2-like receptor efficacy is directly related to efficacy as a reinforcer. It is possible that other pharmacological effects, e.g., D1 receptor activity, influenced self-administration.     

Dopamine D2/D3 receptors modulate cocaine's reinforcing and discriminative stimulus effects in rhesus monkeys

Numerous studies have suggested that dopamine (DA) D2 and D3 receptors are involved in the behavioral effects of cocaine. The present experiments evaluated the reinforcing and cocaine-like discriminative stimulus effects of several D2/D3 agonists in rhesus monkeys. In the first experiment, animals (n = 4) were trained to self-administer 0.03 mg/kg/inj cocaine under a fixed-interval (FI) 5-min schedule. When substituted for cocaine, the D2/D3 agonist quinpirole (0.003-0.03 mg/kg/inj) functioned as a reinforcer in all monkeys. In two cocaine-naive monkeys trained to respond under an FI 3-min schedule of food presentation, quinpirole maintained low rates of responding in one subject, while at the highest dose (0.03 mg/kg/inj) it functioned as a reinforcer in the second monkey. In this animal, increased activity was observed at this dose, which may have contributed to the overall rate of responding. In the second experiment, monkeys (n = 4) were trained to discriminate cocaine from saline using a two-lever, food-reinforced, drug discrimination procedure. The D2/D3 agonists quinpirole, (+/-)-7-OH-DPAT, and R-( + )-7-OH-DPAT fully substituted for cocaine. However, the time-course of substitution differed between quinpirole, which substituted for cocaine 10 min after administration, and (+/-)- and R-(+)-7-OH-DPAT, which required 60-min pretreatments. The behavioral potencies, as determined from ED50, values, correlated with previously reported in vitro binding affinity and functional activity at the D3 receptor [R-(+ )-7-OH-DPAT > (+/-)-7-OH-DPAT > quinpirole]. These results further indicate that direct-acting D2/D3 agonists can function as reinforcers and produce cocaine-like discriminative stimulus effects, and support the idea that D3 receptors should continue to be a valuable target for future behavioral studies evaluating cocaine's mechanisms of action.     

Relationship between receptor occupancy and response at striatal dopamine autoreceptors

The irreversible dopamine (DA) receptor antagonist N-ethoxy-carbonyl-2-ethoxy-1,2-dihydroquinoline (EEDQ) was used to determine the extent of receptor reserve at DA autoreceptors regulating in vivo tyrosine hydroxylase activity. Rats were treated with vehicle or EEDQ (1 X 0.5-2 X 6 mg/kg, subcutaneously) and, 24 hr later, dose response curves were generated for DA agonist reversal of gamma-butyrolactone-induced striatal L-3,4-dihydroxyphenylalanine (L-DOPA) accumulation. Double reciprocal plots were obtained of equieffective doses of agonist required to elicit response at several levels of effect before and after partial irreversible receptor inactivation. A pseudo-dissociation constant (pseudo-KA, in units of dose) and the fraction of receptors remaining active (q) were determined; these values were then used to calculate the relationship between receptor occupancy and response. The ED50 (1 microgram/kg) for the full DA receptor agonist N-propylnorapomorphine (NPA) was shifted 2.8, 4.8-, and 11.3-fold to the right after partial irreversible receptor blockade which left the fraction of receptors remaining active (q) at 0.37, 0.17 and 0.058, respectively. Corresponding maximal reversal of L-DOPA accumulation was 100, 77, and 58%, indicating a nonlinear relationship between receptor occupancy and response for NPA and the presence of a large receptor reserve; maximal and half-maximal responses were calculated to require occupancy of 30 and 3.8% of the total receptor pool, respectively. Dose response curves were also obtained for the DA autoreceptor-selective agents EMD 23,448 and (+)- and (-)-3-PPP before and after EEDQ treatment. In controls, EMD 23,448 and (+)-3-PPP, like NPA, completely reversed striatal gamma-butyrolactone-induced L-DOPA accumulation, whereas the maximal effect of (-)-3-PPP was 52% reversal. After EEDQ treatment (6 mg/kg), EMD 23,448 and (+)-3-PPP showed relatively small shifts in ED50 values. Furchgott analysis demonstrated that all three atypical agents are partial agonists at the DA autoreceptor with efficacies of 0.19 (EMD 23,448), 0.12 [(+)-3-PPP], and 0.05 [(-)-3-PPP] relative to NPA. The presence of a larger receptor reserve at pre-versus postsynaptic D2 DA receptors and the partial agonist character of drugs such as EMD 23,448 and the enantiomers of 3-PPP may account for their autoreceptor selectivity.     

Hypothermia in mice: D2 dopamine receptor mediation and absence of spare receptors

The nonselective dopamine (DA) receptor agonists R(-)apomorphine (APO) and R(-)-N-n-propylnorapomorphine (NPA) elicited dose- and time-dependent hypothermia in mice with ED50 values of 300 and 18 micrograms/kg, respectively. The selective D2 agonist quinpirole (LY 171555) also elicited dose-dependent hypothermia, whereas the selective D1 agonist SKF 38393 had no effect. The selective D1 and D2 antagonists SCH 23390 (1 mg/kg) and sulpiride (200 mg/kg), respectively, did not significantly alter body temperature. The hypothermic effect of a maximal dose of NPA (0.2 mg/kg) was not blocked by SCH 23390 (1 mg/kg) but was significantly attenuated (p less than 0.001) by pretreatment with sulpiride (200 mg/kg). Pretreatment with sulpiride (200 mg/kg) produced a parallel, 40-fold shift to the right of the dose-response curve for NPA. Partial, irreversible DA receptor inactivation by N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline (EEDQ) (2 mg/kg) reduced the maximal hypothermic effect of NPA (to 49% of control) without altering its ED50. Analysis of the data indicated a linear relationship between DA receptor occupancy and hypothermic response. The results demonstrate that DA agonist-induced hypothermia in mice is mediated by D2 receptors and that there is no receptor reserved for this response.     

The role of the dopaminergic system in the modulation of the acoustic startle response in the rat

The (+) and (-) isomers of 3-(3-hydroxyphenyl)-N-propylpiperidine (3-PPP) (2-32 mg/kg) were tested for their effects on the acoustic startle response in the rat and these effects were compared with those of apomorphine and haloperidol. Both startle amplitude and startle latency were recorded. (+)-3-PPP produced a dose-dependent prolongation of startle latency. No other significant effects of the 3-PPP isomers were observed. Apomorphine (0.75-3 mg/kg) produced a dose-dependent increase in startle amplitude concurrent with a dose-dependent prolongation of startle latency. Both these effects were blocked by haloperidol. Haloperidol by itself produced a significant decrease in startle amplitude at the highest dose used (1.6 mg/kg). These and other data suggest that activation of both dopamine D-1 and dopamine D-2 receptors is essential for the stimulatory effects of dopamine receptor agonists on the acoustic startle response in the rat.     

Behavioral effects of apomorphine isomers in the rat: Selective locomotor-inhibitory effects of S(+)N-n-propylnorapomorphine

The optical isomers of apomorphine (APO) and N-n-propylnorapomorphine (NPA) were evaluated behaviorally in the rat. Both R(-) isomers induced motor-excitatory effects and strong stereotyped sniffing, licking, and gnawing, as has been reported previously. The S(+) isomers selectively inhibited locomotor activity and did not induce stereotypy or catalepsy. These actions of the S(+) aporphines were selective against locomotor activity stimulated by low doses of R(-) isomers. (+)NPA (ID₅₀=0.2 mg/kg) was 20 times more potent than (+)APO (ID₅₀=4 mg/kg) in antagonizing the locomotor arousal-inducing effects of (-)APO (at ED₅₀=0.3 mg/kg). (+)NPA also inhibited spontaneous locomotor activity much more potently (ID₅₀=3.0 mg/kg) than did (+)APO (ID₅₀>50 mg/kg). Neither S(+) aporphine had a significant effect against stereotypy induced by the R(-) isomers, even at high doses (up to 30 mg/kg). Inhibition of the effects of (-)APO by (+)NPA appeared not to be due to altered uptake of (-)APO into brain. These results suggest that S(+)NPA or its congeners and analogs may have selective antidopaminergic actions in limbic rather than striatal areas of mammalian brain.     

Effects of isomers of hydroxyaporphines on dopamine metabolism in rat brain regions

The effects of isomers of di- and monohydroxyaporphines on cerebral dopamine (DA) metabolism were evaluated in representative extrapyramidal (corpus striatum) and limbic (nucleus accumbens septi) tissues of rat brain by three methods: (1) changes in the ratio of homovanillic acid (HVA) to DA, (2) accumulation of L-dihydroxyphenylalanine (DOPA) after inhibiting its decarboxylation to DA under "open-loop" conditions, as well as (3) after gamma-butyrolactone (GBL) pretreatment to provide selective effects at presynaptic DA autoreceptors. The DA-agonist R(-) isomers of the aporphines apomorphine (APO), N-n-propylnorapomorphine (NPA), and 11-hydroxy-N-n-propylnoraporphine (11-OH-NPa) showed consistent dose-dependent inhibition of DA synthesis in both brain regions with all models; the neuroleptic haloperidol had the opposite effect in the first two models only, as expected. The S(+) isomers of NPA and 11-OH-NPa have shown behavioral evidence of antidopaminergic activity, especially in the limbic system. Unlike the neuroleptic, S(+)NPA did not show DA-synthesis enhancing actions in accumbens or striatal tissue but, instead, inhibited DA synthesis like its R(-) antipode in all three test paradigms. S(+)11-OH-NPa given alone produced minor changes in the HVA/DA ratio and did not antagonize R(-)11-OH-NPa, weakly increased accumulation of DOPA in the second model, and had no effect in the third--all without regional selectivity. In the test of autoreceptor functioning, the dihydroxyaporphine S(+)NPA, but not S(+)11-OH-NPa, inhibited DA synthesis and this effect, in turn, was largely reversed by haloperidol, as were the inhibitory effects of the three R(-)aporphines tested. In this model, however, neither S(+)NPA nor S(+)11-OH-NPa antagonized the DA-synthesis inhibiting effect of R(-)APO as haloperidol did. Overall, these results are consistent with evidence that R(-)NPA and 11-OH-NPa have high affinity at D-2 receptor sites in rat brain and show behavioral effects of typical DA agonists. The non-stereoselective inhibitory effects of NPA on DA synthesis may reflect its activity as a weak DA agonist with very low intrinsic activity, but may also include a direct "catechol-effect" on tyrosine hydroxylase. In contrast, R(-)11-OH-NPa appears to be a stereoselective D-2 agonist, active at autoreceptors as well as postsynaptic receptors, that lacks the nonstereospecific effects on DA metabolism of its catechol-aporphine congener. It may be a useful probe for the further characterization of dopamine receptors and autoreceptors.     

(+)- and (-)-3-PPP exhibit different intrinsic activity at striatal dopamine autoreceptors controlling dopamine synthesis

Both enantiomers of the dopamine analogue 3-(3-hydroxyphenyl)-N-n-propylpiperidine (3-PPP; 0.5-32 mg/kg s.c.) dose dependently reduced the increase in striatal dopamine (DA) synthesis rate produced by gamma-butyrolactone (GBL). Whereas (+)-3-PPP completely prevented the action of GBL, (-)-3-PPP was only partially effective. In addition, (-)-3-PPP partially antagonised the inhibitory action of apomorphine on the GBL-induced increase in DA synthesis rate. These findings suggest that (+)- and (-)-3-PPP act as full and partial agonists respectively, at striatal DA autoreceptors controlling DA synthesis.     

Modulation of cocaine's discriminative stimulus effects by dopamine D(1) agonists in rhesus monkeys

Dopamine (DA) D(1) agonists are classified as high- or low-efficacy on the basis of in vitro functional measures as compared to DA. In monkeys self-administering cocaine, high-efficacy D(1) agonists have been shown to have reinforcing effects, while low-efficacy agonists do not. However, the relationship between D(1) agonist efficacy and cocaine-like discriminative stimulus effects, particularly in rhesus monkeys, is not clear. The present study investigated the discriminative stimulus effects of a high- (SKF 81297) and a low-efficacy (SKF 38393) D(1) agonist in rhesus monkeys (n=4) trained to discriminate cocaine from saline using a two-lever drug discrimination procedure. In a second experiment, the effects of agonist pretreatments, as well as pretreatment with a D(1) antagonist, on cocaine's discriminative stimulus effects were evaluated. SKF 81297 (0.01-1.7 mg/kg) fully substituted for cocaine in three of four animals (> 80% cocaine-appropriate responding), while SKF 38393 (0.3-10 mg/kg) occasioned < 50% cocaine-appropriate responding in all subjects. When given as a pretreatment, neither agonist altered cocaine's discriminative stimulus effects at the doses tested. In contrast, the D(1) antagonist SCH 23390 attenuated cocaine's discriminative stimulus effects. These results indicate that D(1) agonists have cocaine-like discriminative stimulus effects in rhesus monkeys that are consistent with their in vitro efficacies. However, when given in combination with cocaine, D(1) agonist efficacy does not appear to be a major factor in modifying cocaine's discriminative stimulus effects.     

Evidence for a role for dopamine D3 receptors in the effects of dopamine agonists on operant behaviour in rats

Several dopamine (DA) agonists have been reported to show some D3 versus D2 selectivity but the extent of this selectivity depends on experimental conditions, and the behavioural effects of these compounds seem to differ little from those of non-selective agonists such as apomorphine. However, some recent studies have reported stronger correlations between several behavioural responses and D3 affinities than between the same responses and affinities for D2 receptors. In the present study rats were trained to lever press for food on a fixed-ratio (FR10) schedule during daily 15min sessions. The DA agonists apomorphine; quinelorane; quinpirole; 7-hydroxy-2-(di-n-propylamino)-tetralin [(+/-)7-OH-DPAT]; (+)-(4aR, 10bR)-4-propyl-3, 4,4a, 10b-tetrahydro-2H,5H-1-benzopyrano [4,3-b], 4(oxazin-9-ol); bromocriptine; and (3-hydroxyphenyl)-N-propylpiperidine produced dose-related decreases in these response rates. The potencies of these compounds correlated significantly with their published potencies to produce a functional D3 but not a functional D2 response (stimulation of mitogenesis in transfected cells). The rate-decreasing effects of 7-OH-DPAT were antagonised by the benzamide antipsychotic agent amisulpride, at low doses (1 and 3mg/kg) which have been shown to exert preferential activity at presynaptic DA receptors. Haloperidol and remoxipride produced only small antagonist effects. These results are consistent with the view that D3 DA receptors may play an important role in mediating the behavioural effects of DA agonists and that these receptors have a presynaptic location.     

Effects of N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline on the prolactin suppression induced by a series of full and partial dopamine D2 receptor agonists in male rats

Summary The effect of pretreatment with a high dose of the alkylating agent N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline (EEDQ) (20 mg/kg, 24 h) on the intrinsic activity displayed by a series of full and partial dopamine D2 receptor agonists on prolactin regulating pituitary D2 receptors in male rats was studied. To increase baseline prolactin levels, gamma-butyrolactone in a dose inhibiting brain dopamine neurotransmission was given to all animals. In controls, i. e. rats not given EEDQ, supramaximal doses of all full and partial D2 receptor agonists tested decreased serum prolactin levels with > 80%. While the intrinsic activities of the dopamine precursor 1-DOPA and of the full agonists (+)-3-PPP, 5-OH-DPAT, B-HT 920 (talipexole), apomorphine, and NPA (R-(–)-N-n-propylnorapomorphine) were not affected by pretreatment with EEDQ, the effects of supramaximal doses of the partial agonists (–)-HW-165, TDHL (terguride), SDZ 208-911, (–)-3-PPP (preclamol), and SDZ 208–912 were reduced to 66%, 74%, 59%, 100%, and 100%, respectively. The effect of EEDQ on the intrinsic activity displayed by the various agonists corresponds inversely to the intrinsic efficacy displayed by the drugs in other models of D2 receptor function with one exception only; thus, the prolactin suppressive effect of (–)-3-PPP was more effectively antagonized by EEDQ than would have been predicted from the intrinsic efficacy usually attributed to the drug. Since the dose of EEDQ used in the present study has previously been shown not to decrease D2 receptor density in the pituitary as measured using in vivo radioligand binding, it is suggested that alkylation of D2 receptors may change the conformation of the individual receptor complexes in a way that decreases the responsiveness to partial but not full agonists. Send offprint requests to A. Ekman at the above address     

Efficacy and potency comparisons among aporphine enantiomers: effects on dopamine neurons in substantia nigra of rat

Extracellular single unit recording studies were carried out on male rats to determine the responses of dopamine neurons of the substantia nigra to intravenous administration of the enantiomers of the aporphine congeners, apomorphine (APO), N-n-propylnorapomorphine (NPA) and 11-hydroxy-N-n-propylnorapomorphine (11-OH-NPa). The R-(-)-configuration was found to be the most critical determinant of the efficacy and potency of the agonists. All R-(-)-aporphines were full agonists, able to inhibit completely firing of dopamine cells. The order of potencies, defined by the ID50s, was: (-)NPA, 2.0 +/- 0.4 nmol/kg greater than (-)11-OH-NPa, 4.7 +/- 0.7 nmol/kg greater than (-)APO, 18.0 +/- 4.0 nmol/kg. Thus, potency was increased about 9-fold by replacing the 6N methyl of APO with an n-propyl (NPA). Conversely, the 10-hydroxy was not essential for agonist activity (11-OH-NPa) but could increase potency. In the S-(+)-series responses varied. (+)N-n-Propylnorapomorphine exhibited agonist properties and could fully inhibit dopamine cells, but its potency was low (ID50 1550 nmol/kg); (+)APO produced only slight but significant decreases in firing at large (8434 nmol/kg) doses and (+)11-OH-NPa was devoid of efficacy in that it caused no significant changes in firing. Despite their loss of efficacy and potency, the (+)-enantiomers apparently did retain affinity for DA receptors, since they could act as antagonists if given before (-)APO or NPA. These findings suggest that stereochemical conformation and key structural elements of the aporphines are interactive in determining agonist efficacy and potency within this physiological test system.     

The effects of dopamine D3 agonists and antagonists in a nonhuman primate model of tardive dyskinesia

Tardive dyskinesia (TD), a serious complication of antipsychotic dopamine (DA) antagonist treatment, has been hypothesised to develop due to a dominant DA D1 relative to DA D2 receptor function. Recent genetic and pharmacological studies implicate the DA D3 receptor in TD. The present study examined the role of the DA D3 receptor in relation to the DA D1/D2 imbalance hypothesis of TD in nonhuman primates. Eight Cebus monkeys displaying mild to severe TD due to previous chronic exposure to DA D2 antagonists were acutely injected with SKF 81297 (DA D1 agonist) 0.3 and 0.6 mg/kg, pramipexole (DA D3>D2 agonist) 0.025-0.1 mg/kg, CIS-8-OH-PBZI (DA D3 agonist) 5-10 mg/kg and SB-27701-A (DA D3 antagonist) 1-5 mg/kg and rated for oral dyskinesia. SKF 81297, 0.3 and 0.6 mg/kg, exacerbated TD. Pramipexole and CIS-8-OH-PBZI reduced SKF 81297-induced TD, while SB-27701-A had no effect. When administered alone, SB-27701-A increased TD relative to placebo, while pramipexole and CIS-8-OH-PBZI had no significant effect. Pramipexole did, however, ameliorate TD in those monkeys with severe TD. These results point towards a role of the DA D3 receptor in TD, but indicate that the DA D2 receptor may also play an essential role.     

R(-) and S(+) stereoisomers of 11-hydroxy- and 11-methoxy-N-n-propylnoraporphine: central dopaminergic behavioral activity in the rat

R(-)11-Hydroxy-N-n-propylnoraporphine (11-OH-NPa) induced stereotyped behavior in the rat as potently (ED50 = 0.80 mg/kg, i.p.) as R(-)apomorphine (APO) and this effect was blocked by haloperidol; the 11-methoxy congener, R(-)11-MeO-NPa, had a weak effect (ED50 greater than 10 mg/kg) and the S(+) isomers had none. The isomer R(-)11-OH-NPa potentiated locomotion stimulated by apomorphine; S(+)11-OH-NPa inhibited it and the isomers of 11-MeO-NPa were inactive. Catecholaporphines usually are inactive orally, but both R(-) and S(+)11-OH-NPa were similarly potent after oral or parenteral administration. The isomer S(+)11-OH-NPa inhibited spontaneous and apomorphine-induced locomotion (ID50 = 1.8-2.7 mg/kg, p.o. and i.p.) and stereotyped behavior (ID50 = 3 mg/kg, p.o. or i.p.), all without inducing catalepsy. While apomorphine was short-acting (1-2 hr), the effects of R(-)11-OH-NPa persisted up to 6-7 hr and those of the S(+) isomer for at least 2.5 hr; moreover, the efficacy of R(-)11-OH-NPa increased markedly up to 3-4 hr, although its ED50 was independent of time (ED50 = 1.7-1.9 mg/kg, i.p. from 1-3 hr). The total effect of R(-)11-OH-NPa (p.o. or i.p.) over time was more than 10-times greater than that of injected apomorphine. These observations accord with the reported high (nM) affinity of 11-OH-NPa at cerebral DA receptor sites (D2 greater than D1) and weak interactions of the 11-methoxy congener. They support the conclusion that the R(-) and S(+) stereoisomers are neuropharmacologically active, respectively, as DA agonist and apparent antagonist, as was found with the enantiomers of N-n-propylnorapomorphine, perhaps due to the low intrinsic postsynaptic agonist activity of the S(+) isomers. Moreover, 11-OH-NPa was highly bioavailable orally and unusually long-acting; it may be absorbed slowly or have active metabolites. Hydroxy-substitution of aporphines at the 11-position, homologous to the 3-OH of DA, evidently is critical for affinity and activity at the DA receptor. These or other monohydroxyaporphines may represent leads to potentially useful DA agonist or antagonist drugs.     

Dopamine autoreceptor agonists attenuate spontaneous motor activity but not spontaneous fighting in individually-housed mice

The present study was conducted to determine whether or not two behavioral characteristics of individually-housed mice, hyperactivity in a novel environment and intermale fighting, are attenuated by the dopamine (DA) agonists, apomorphine, (+)- and (-)-3-(3-hydroxyphenyl)-N-n-propylpiperidine (3-PPP). Autoreceptor-activating doses of these drugs which reduced spontaneous activity in a novel environment did not inhibit spontaneous fighting with conspecific olfactory bulbectomized males. Individually-housed mice were more active in a novel environment and showed a significant reduction of activity at lower doses of apomorphine, (+)- and (-)-3-PPP than group-housed mice. However, the ED50's for the inhibition of spontaneous activity in a novel environment in group- and individually-housed mice were similar: apomorphine, 0.02 vs. 0.012 mg/kg, SC; (+)-3-PPP, 0.50 vs. 0.51 mg/kg, SC; and (-)-3-PPP, 1.0 vs. 0.56 mg/kg, SC, for group- and individually-housed mice respectively. A significant proportion of individually-housed mice, but not group-housed mice, displayed catalepsy in response to high doses of (-)-3-PPP. These data suggest that DA autoreceptor agonists can modulate the hyperactivity syndrome but not spontaneous fighting behavior in individually-housed mice.     

Dopamine D2 autoreceptors in rats are behaviorally functional at 21 but not 10 days of age

Previous studies used either racemic 3-(3-hydroxyphenyl)-N-n-propylpiperidine [(±)-3-PPP] or lower doses of the mixed dopamine (DA) D₁/D₂ agonist apomorphine (APO) to conclude that brain DA D₂ autoreceptors are not behaviorally functional until 28 days of age. The purpose of this study was to provide behavioral evidence for functional D₂ autoreceptors before 28 days of age using DA agonists with greater selectivity for D₂ autoreceptors. The locomotor activity of 10-, 21-, 35-day-old and adult rats was monitored after injection of a D₂ autoreceptor agonist. There were significant decreases in the locomotor activity of 21-, 35-day-old, and adult rats injected with (-)-3-PPP, SND 919, or PD 128483. Lower doses of APO significantly decreased the activity of adult and 35-day-old rats but not younger rats. The only significant effect on the locomotor activity of 10-day-old rats was an increase in activity after injection of APO, 0.01 mg/kg or higher, or B-HT 920, 0.01 mg/kg. The results suggest that brain DA D₂ autoreceptors are behaviorally functional at 21, but not 10, days of age.     

Agonist and antagonist effects of 3-PPP enantiomers on functional dopamine autoreceptors and postsynaptic dopamine receptors in vitro

In contrast to racemic 3-PPP (3-(3-hydroxyphenyl)-N-n-propylpiperidine), (+)-3-PPP appeared to inhibit the electrically evoked release of both [3H]dopamine (DA) and [14C]acetylcholine (ACh) from superfused rat neostriatal slices, although it was considerably less potent in this respect that the DA receptor agonists apomorphine, TL-99 (6,7-dihydroxy-N,N-dimethyl-2-aminotetralin) and LY 141865. At concentrations higher than 1 microM both of the 3-PPP enantiomers increased the spontaneous efflux of 3H but not that of 14C. (+)3-PPP also inhibited the cholera toxin-stimulated release of immunoreactive alpha-MSH from dispersed intermediate lobe cells of the rat pituitary gland. The inhibitory effects of (+)3-PPP on both transmitter and alpha-MSH release were antagonized by the selective D-2 receptor antagonist (-)-sulpiride. Neither [3H]DA nor [14C]ACh release were inhibited by (-)3-PPP but, in contrast, the release-inhibiting effect of the selective D-2 receptor agonist LY 141865 as well as that of (+)3-PPP were antagonized by (-)3-PPP, although less effectively than by (-)sulpiride. The inhibitory effect of LY 141865 on alpha-MSH release from intermediate lobe cells was also antagonized by (-)3-PPP. The data indicate that (+)3-PPP is a weak agonist and (-)3-PPP a weak antagonist at D-2 receptors and that neither of the 3-PPP enantiomers interacts selectively with DA autoreceptors mediating presynaptic modulation of striatal DA release.     

Dopamine D2 receptor knock-out mice are insensitive to the hypolocomotor and hypothermic effects of dopamine D2/D3 receptor agonists.

The dopamine (DA) D2-like family of receptors is comprised of three subtypes, the D2, D3, and D4 receptors. It has been suggested that the potency of DA receptor agonists to produce hypothermia and hypolocomotion in rodents correlates more strongly with the in vitro affinity for, or potency (mitogenesis test) at the D3 than at the D2 subtype. However, it has recently been reported that when tested in DA D3 receptor knock-out mice, several DA D2/D3 receptor agonists (7-OH-DPAT, PD 128907 and quinelorane) induced levels of hypothermia and decreases of locomotor activity similar to those obtained in control (wild-type) mice. These results do not argue in favour of an implication of DA D3 receptors in these in vivo effects. In order to investigate whether the DA D2 receptor is the subtype that mediates hypothermia and hypolocomotion produced by DA D2/D3 receptor agonists, we tested the effects of ip administration of the DA D2/D3 receptor agonists 7-OH-DPAT and PD 128907, on core temperature and locomotor activity in DA D2 receptor knock-out mice (homozygotes: D2(-/-) and heterozygotes: D2(+/-)), and in wild-type (D2(+/+)) mice. 7-OH-DPAT (0.1-3 mg/kg) and PD 128907 (1-10 mg/kg) induced hypothermia and decreased locomotion in D2(+/+) mice, but had no effects in D2(-/-) mice; the magnitude of the hypothermic and locomotor-reducing effects of these two agonists in D2(+/+) mutants was approximately half that of D2(+/+) mice. During the first 10 min in the activity chambers, the level of spontaneous locomotor activity of D2(-/-) individuals was almost 50% below that of D2(+/+) mice; basal locomotor activity of D2(+/-) mice was between that of D2(-/-) and D2(+/+) individuals. Neither type of mutant showed spontaneous catalepsy or deficits in forelimb muscle strength (grip-strength test). These results show that the presence of DA D2 receptors is necessary for the expression of the locomotor- and core temperature-decreasing effects of DA D2/D3 receptor agonists such as 7-OH-DPAT and PD 128907.     

Occurrence of yawning and decrease of prolactin levels via stimulation of dopamine D2-receptors after administration of SND 919 in rats

Summary SND 919 ((S)-2-amino-4,5,6,7-tetrahydro-6-propylamino-benzothiazole) is expected to have a potent and selective dopamine D₂-receptor agonistic activity. From this information, the present study was performed to investigate effects of SND 919 on yawning behavior and prolactin secretion in rats. Subcutaneous injections of SND 919 (25–500 gm/kg, s. c.) elicited yawning responses. Its dose-response curve was bell-shaped with maximal effects at a dose of 100 g/kg. Yawning behavior was also evoked by the putative dopamine autoreceptor agonists, talipexole (6-allyl2-amino-5,6,7,8-tetrahydro-4H-thiazolo [4,5-d]azepine) (BHT 920) (5–100 g/kg, s. c.) and (+)-3-PPP ((+)-3-(3-hydroxyphenyl)-N-n-propylpiperidine) (5–15 mg/kg, s. c.). The yawning induced by SND 919 (100 g/kg, s. c.) as well as talipexole (25 g/kg, s. c.) was inhibited by pretreatment with dopamine D2-receptor antagonists such as spiperone (0.5 mg/kg, i.p.) and YM-09151-2 (cis-N-(1-benzyl-2methylpyrrolidin-3-yl)-5-chloro-2-methoxy-4-methylaminobenzamide) (0.1 mg/kg, i. p.), or the muscarinic receptor antagonist, scopolamine (0.5 mg/kg, i.p.). However, the yawning was not affected by the dopamine D₁-receptor antagonist, SCH 23390 (R(+)-8-chloro-2,3,4,5-tetrahydro-3methyl-5-phenyl-1 H-3-benzazepine-7-ol) (0.5 mg/kg, i. p.). Stereotypy such as licking and biting was not observed following the administration of SND 919, talipexole and (+)-3-PPP. Administration of SND 919, talipexole or (+)-3-PPP in respective yawn-inducing doses caused a reduction in both the basal prolactin levels and the a-methyl-p-tyrosine-induced hyperprolactinemia. The results suggest that SND 919 as well as talipexole exerts selective agonistic activities for specific dopamine D2-receptors, which are not related to the occurrence of stereotypy and have a high affinity for dopamine receptor agonists quite similar to that of the pituitary lactotroph dopamine D₂-receptors.Send offprint requests to K. Yamada at the above address     

Dopamine D-2 receptor agonist-induced behavioural depression: critical dependence upon postsynaptic dopamine D-1 function

Summary The dopamine (DA) D-2 receptor agonists quinpirole (threshold dose, 0.01 mg/kg IP), pergolide (0.025 mg/kg), B-HT 920 (0.003 mg/kg) and (–)-3-PPP (4 mg/kg) produced dose-dependent locomotor depression (immobility) in mice as assessed by a subjective scoring system, with the immobility being characterized by a frozen posture. The animals were still but had their eyes open. The immobility was accompanied by reductions in sniffing, rearing and grooming. The depression (and the associated reduction in the various behaviours) produced by quinpirole (0.1 mg/kg), pergolide (0.1 mg/kg) and B-HT 920 (0.1 mg/kg) was substantially (but not always completely) reversed by the selective D-1 receptor agonist SKF38393 (up to 12 mg/kg) and the non-selective D-1 receptor agonist CY208243 (up to 3 mg/kg). The immobility induced by (–)-3-PPP (16 mg/kg) was also reversed by CY208243 and SKF38393, but the reversal was due to an increase in grooming behaviour in mice challenged with the D-1 receptor agonists, whether or not the animals had also received (–)-3-PPP. There was no reversal of the depression of rearing or sniffing. In contrast, CY208243 and SKF38393 also antagonized the immobility induced by B-HT 920, but the reversal was accompanied by at least partial reversals of the depression of sniffing, rearing and grooming. The reversal of quinpirole-induced immobility by SKF38393 and CY208243 was antagonized by SCH23390 (0.1 mg/kg). The selective D-2 receptor antagonist raclopride (0.025 to 0.4 mg/kg) could not reverse quinpirole-induced immobility. High doses of either raclopride (0.4 mg/kg) or SCH23390 (> 0.1 mg/kg) significantly increased immobility. Although raclopride itself (0.2 mg/kg) produced a substantial increase in DOPAC and homovanillic acid (HVA) levels in the striatum, it did not antagonize the autoreceptor mediated effects of quinpirole (0.1 mg/kg) in reducing the striatal dihydroxyphenylacetic acid (DOPAC) to DA ratio. However, the same dose of raclopride was partly effective in reducing the effects of lower doses of quinpirole (0.01 and 0.03 mg/kg) on the striatal DOPAC to DA ratio. Raclopride (0.2 mg/kg) also partially but significantly reduced the locomotor stimulant effects of d-amphetamine in reserpinized mice. Biochemical analyses in the striata indicated that CY208243 slightly retarded DA turnover (as assessed by the DOPAC/DA ratio). SKF38393 itself also slightly reduced DA turnover. In automated activity cages, using mice depleted of DA with reserpine and a-methyltyrosine, all the D-2 receptor agonists tested, in combination with SKF38393, produced an increase in activity. It is concluded that the depression induced by D-2 receptor agonists is due substantially to a deprivation of DA at postsynaptic D-1 receptors and that, at the doses tested, the D-2 receptor agonists were able to exert measurable stimulation of the postsynaptic receptors. The ability of D-1 receptor agonists to reverse the D-2-mediated depression is due to the stimulation of the postsynaptic D-1 receptors with the injected D-1 receptor agonist (replacing the endogenous transmitter) and the stimulation of the postsynaptic D-2 receptors by the injected D-2 receptor agonist.