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Nucleus accumbens dopamine and discriminated approach learning: interactive effects of 6-hydroxydopamine lesions and systemic apomorphine administration.

RATIONALE: Although dopaminergic mechanisms have been implicated in incentive motivational processes, their role in appetitive conditioning remains poorly understood. OBJECTIVES: To investigate the effects of dopamine (DA) depleting lesions of the nucleus accumbens and the direct acting dopamine receptor agonist apomorphine on the learning of discriminated appetitive approach behavior in a Pavlovian autoshaping paradigm. METHODS: Rats received bilateral infusions of either phosphate-buffered saline (shams) or 6-hydroxydopamine (lesions) directly into the nucleus accumbens. Ten days later, rats were trained on an autoshaping task whereby a previously neutral light stimulus was paired with food reward. Presentation of another stimulus (CS-) was never followed by reward. Over 100 pairings subjects developed a conditioned response of approaching the reward-predictive stimulus (CS+). Prior to each autoshaping session subjects were administered either saline (1 ml/kg SC) or apomorphine (30 microg/kg; 100 microg/kg), in a between-subjects design. RESULTS: Lesioned subjects showed a delay in the acquisition of discriminated approach and were insensitive to a subsequent change in the contingency of the task whereby approaches to the CS+ now prevented food delivery. Low dose apomorphine profoundly impaired learning in both sham and lesioned subjects. Despite increasing the overall number of CS+ and CS- approaches, the higher dose of apomorphine allowed discriminated approach in sham-operated animals only. CONCLUSIONS: These data are compatible with the hypothesis that nucleus accumbens dopamine serves to energize rather than guide conditioned approach to appetitive cues. They also support the notion that DA inputs in this region confer flexibility of approach to cues predictive of reward.

Animals↗

Repeated apomorphine administration alters dopamine D1 and D2 receptor densities in pigeon basal telencephalon.

When pigeons are repeatedly administered a dose of apomorphine they show an increasing behavioral response, much as rodents do. In birds this expresses itself in an augmented pecking response. This sensitization is assumed to be largely due to a conditioning process. Here we present evidence that sensitization is accompanied by an alteration of the D(1) to D(2) dopamine receptor densities. An experimental group of pigeons was repeatedly injected with apomorphine, and a control group with saline. The basal forebrain tissue, known to be rich in dopamine receptors, was subjected to binding assays using tritiated specific D(1) and D(2) dopamine receptor antagonists. There was a trend towards an increase in D(1) and a significant decrease in D(2) receptor densities in apomorphine-treated birds compared to the saline-treated controls. We conclude that extended apomorphine treatment modifies the D(1) dopamine receptor density in the opposite manner to the D(2) dopamine receptor density.

Animals↗

Key issues from the clinical trials of apomorphine SL.

The central nervous system has the capacity to enhance the activity of dysfunctional penile tissue in men with erectile dysfunction (ED). Phase III clinical trials have been conducted using Apomorphine SL (TAP Pharmaceuticals, Deerfield, IL) as a centrally acting treatment for ED. Apomorphine SL has been administered to over 3,000 men in over 75,000 doses. In three phase III crossover double blind studies 854 patients were given a total of 8,263 tablets of apomorphine SL in 2 and 4 mg doses. The patients were between 18 and 70 years old and had multiple co-morbid conditions. Outcome measures included intercourse rates and erection rates on a per attempt basis as well as psychometric instruments and partner response evaluations. The results show that 74.1% of patients had moderate or severe grades of ED on inclusion into the studies, 31% had hypertension, 16% had documented coronary artery disease, 16% had dyslipidemia, and 16% had diabetes. Erections occurred rapidly (10-25 min). In 54.4% of attempts at 4 mg (vs 33.8% placebo, P < 0.001) erections suitable for intercourse were documented. A majority of the attempts at intercourse (50.6%, P < 0.001) were successful at 4 mg a doubling of baseline rates. Mild nausea was the most common but infrequent side effect and the rare occurrence of syncope was the most significant. No cardiac deaths were attributed. It is concluded that the clinical trials of apomorphine SL demonstrate a safe and significant rate of restoration of erectile function by means of a central mode of action. Efficacy has been shown in men with cardiovascular disease and severe grades of ED.

Adolescent↗

Growth hormone response in low-dose apomorphine test correlates with nigrostriatal dopamine transporter binding in patients with Parkinson's disease.

Challenge with low-dose apomorphine causes a rise in growth hormone (GH) in patients with Parkinson's disease (PD). We studied 18 patients with early PD, who showed an increase of GH in the low-dose apomorphine test, by means of [(123)I] FP-CIT-SPECT. The mean specific dopamine transporter binding of the 18 patients was 1.50 +/- 0.56 in the striatum, 1.20 +/- 0.59 in the putamen, and 1.76 +/- 0.59 in the caudate nucleus. The increase of GH (1.05 +/- 1.01 ng/ml at baseline to 9.46 +/- 6.36 ng/ml 45 min after apomorphine injection; p < 0.001) was significant. There was a significant negative correlation of the increase of GH with the mean specific dopamine transporter binding in all three regions (r between -0.490 and -0.587; p between 0.04 and 0.01). Challenge with low-dose apomorphine may therefore be used as an indirect tool to measure the extent of nigrostriatal neurodegeneration in early PD.

Adult↗

Cholinergic-dopaminergic interaction in the striatum: the effect of 6-hydroxydopamine or pimozide treatment on the increased striatal acetylcholine levels induced by apomorphine, piribedil and d-amphetamine.

Apomorphine (1 and 2 mg/kg), piribedil (15 and 60 mg/kg) and d-amphetamine (5 and 10 mg/kg) increased rat striatal acetylcholine levels without affecting choline. Pretreatment with pimozide (0.5 mg/kg) completely antagonized the effect of apomorphine and piribedil and by itself markedly decreased striatal acetylcholine levels. d-Amphetamine signigicantly antagonized the effect of pimozide. Nine days after pretreatment with 6-hydroxydopamine plus pargyline, striatal dopamine was decreased by 78% while acetylcholine and choline levels remained unaltered. Under these conditions, the effect of d-amphetamine was completely abolished while apomorphine and piribedil were just as active as in the vehicle-treated group. The results suggest that d-amphetamine acted indirectly to increase striatal acetylcholine levels probably through the release of dopamine and/or noradrenaline, while apomorphine and piribedil acted directly at dopamine receptor sites.

Acetylcholine↗

Lesions of substantia nigra by kainic acid: effects on apomorphine-induced stereotyped behaviour.

Fifteen days after bilateral lesions of the substantia nigra by local infusion of kainic acid (0.75 microgram) or after intranigral injection of vehicle, rats were administered 0.1, 0.25, 1.0 and 2.5 mg/kg s.c. of apomorphine and the stereotyped items (locomotion, sniffing and gnawing) were recorded on an event-recorder and motility was measured by a photocell apparatus. After low doses of apomorphine (0.1, 0.2 mg/kg), rats lesioned in the substantia nigra with kainic acid showed a degree of stimulation of motility and of sniffing similar to controls; on the other hand, in rats lesioned with kainic acid in the nigra, a dramatic reduction of gnawing and its replacement by sniffing was observed after administration of higher doses of apomorphine (1.0, 2.5 mg/kg). Bilateral infusion of kainic acid (0.75 microgram) into the reticular information, 2.0 mm dorsal to the substantia nigra, had no effect on apomorphine-induced stereotyped behaviour. These results are in agreement with the concept that the substantia nigra, through non-DA pars reticulata neurons, mediates motor and behavioural syndromes of striatal origin.

Animals↗

Relationships between selective denervation of dopamine terminal fields in the anterior forebrain and behavioral responses to amphetamine and apomorphine.

To investigate the relationship between denervation of dopamine (DA) terminal fields in the anterior forebrain and the behavioral responses to amphetamine (1.5 mg/kg) and apomorphine (1 mg/kg), we injected 6-hydroxydopamine (6-OHDA) bilaterally into the anterolateral hypothalamus (ALH) or into specific mesolimbic and anterior striatal terminal fields after pretreatment with desmethylimipramine to protect noradrenergic axons and terminals from 6-OHDA toxicity. After drug testing was completed, the extent of denervation was determined by fluorescent histochemical analysis. When nearly all of the mesolimbicocortical and anteroventral striatal DA terminal fields were denervated by bilateral ALH 6-OHDA, the locomotor response to amphetamine was abolished, and the locomotor and stereotyped sniffing responses to apomorphine were increased. When fewer DA terminal fields were denervated, different results were obtained: the locomotor response to amphetamine decreased or did not change; stereotyped sniffing elicited by apomorphine did not increase or sniffing was replaced by stereotyped licking and biting. The results suggest a mass action relationship between DA terminal fields in the anterior forebrain and the locomotor response to amphetamine. The relationship between the same DA lesions and responses to apomorphine appears to be more complex.

Amphetamine↗

The effects of chloral hydrate anesthesia on the metabolic response in the substantia nigra to apomorphine.

The effects of apomorphine (1 mg/kg) upon glucose utilization in the substantia nigra (pars compacta) have been examined with 2-deoxyglucose autoradiography in conscious rats and rats anesthetized with chloral hydrate. In conscious rats, apomorphine increased glucose use by 23%, whereas in the anesthetized rats, apomorphine decreased glucose use by 24%. These divergent responses to apomorphine highlight the difficulties associated with the use of chloral hydrate anesthesia in the study of nigro-striatal systems.

Animals↗

Single unit responses of substantia nigra pars reticulata neurons to apomorphine: effects of striatal lesions and anesthesia.

Many of the behavioral consequences of dopamine system activation are thought to be mediated by substantia nigra pars reticulata output pathways. Extracellular, single unit recording studies were conducted to determine how i.v. administration of the dopamine agonist, apomorphine, affects the activity of these pars reticulata neurons. Results revealed that a 320 micrograms/kg dose of the drug, considered sufficient to stimulate striatal postsynaptic dopamine receptors, caused highly variable changes in reticulata cell firing. Cells exhibited increases, decreases, or no changes in firing. Many cells also displayed marked minute to minute changes in firing. This non-uniform pattern of responses was not related to state of consciousness since similar responses were observed in both chloral hydrate-anesthetized as well as conscious, paralyzed rats. Both the increases and decreases could be reversed by subsequent administration of haloperidol. The variable responses to apomorphine were reduced but not totally prevented by striatal kainic acid lesions, suggesting that changes in striatonigral transmission may account for some but not all of the firing changes which were observed. A lower dose of apomorphine (20 micrograms/kg), thought to act primarily at dopamine cell autoreceptors, had little effect on reticulata cell firing and did not modify the variable responses normally observed after the higher dose. These results contrast strikingly with the consistent excitatory responses to apomorphine which have previously been observed in the globus pallidus and suggest that complex or multiple indirect effects of the drug may contribute to the varied reticulata responses.

Anesthesia, General↗

Cerebral glucose utilization following striatal lesions: the effects of the GABA agonist, muscimol, and the dopaminergic agonist, apomorphine.

Local cerebral glucose utilization was measured in 21 discrete regions of the rat CNS following unilateral kainic acid lesions of the caudate nucleus, and subsequent pharmacological challenge with GABAergic or dopaminergic agonists. The most pronounced increases in glucose use were observed in those ipsilateral areas of the brain to which the lesioned striatum normally projects (globus pallidus, entopeduncular nucleus and substantia nigra pars reticulata), although several other brain regions with known anatomical connections to the striato-pallido-nigral system were also affected, most notably in the thalamus and epithalamus. These effects were similar to those reported previously from a different group of animals in which the injection protocol was slightly different. The consequences of striatal lesion, in terms of alterations in local rates of glucose use, were attenuated by i.v. administration of the putative GABA agonist, muscimol. In one area, the ventromedial thalamus, glucose use was more markedly affected by muscimol treatment bilaterally in lesioned animals than in intact animals. The consequences of striatal lesions upon the response to the putative dopaminergic agonist apomorphine, were both complex and profound. In some regions (e.g. globus pallidus), striatal lesion eliminated, or masked the normal response to apomorphine. Elsewhere, the apomorphine response, although in evidence, was significantly attenuated by striatal lesion (e.g. entopeduncular nucleus), but in only two brain areas, substantia nigra pars reticulata and ventrolateral thalamus, the apomorphine response was significantly potentiated by striatal lesion. These studies add further weight to the concept of disinhibition, mediated via striatal GABA fibres, as an organizing principle in striatonigral function and indicate a complex interaction of intrinsic GABAergic pathways with dopaminergic systems in the integrated response to stimulation of dopaminergic receptors in the extrapyramidal motor system.

Animals↗

Activity of A9 and A10 dopaminergic neurons in unrestrained rats: further characterization and effects of apomorphine and cholecystokinin.

The activity of single dopaminergic (DA) neurons (total n = 77) in the midbrain of awake, unrestrained rats was examined. The firing rates and patterns of ventral tegmental area (A10) cells (n = 39) were similar to those of identified DA neurons in anesthetized and paralyzed rats. Unit activity was briefly stimulated (increased firing rate and burst activity) in response to an auditory stimulus and during manual stimulation of the vibrissae. Similar changes occurred during orienting responses and periods of sniffing. Twenty-six percent of A10 cells recorded appeared to be electronically coupled which matched the prevalence previously observed among A9 neurons. The effects on A9 and A10 DA cell activity of apomorphine and the carboxyterminal octapeptide of cholecystokinin (CCK-8) were then determined. Sequential doses of apomorphine (5-320 micrograms/kg, i.v.) reduced the firing rate of each neuron tested (n = 19). Ten of these cells were classified as 'sensitive' to the drug (ED50 less than 20 micrograms/kg), while the remainder were considerably 'less sensitive' (ED50 greater than 30 micrograms/kg). Cells of either sensitivity were as likely to be found in A9 as in A10. Sulfated CCK-8 (1-16 micrograms/kg, i.v.) excited (firing rate and bursting increased) 73% of the A9 neurons sampled but produced inconsistent effects on A10 cell firing. CCK-8 pretreatment increased the percentage of A9 and A10 neurons which were classified as 'sensitive' to apomorphine (82%). Enhanced sensitivity to apomorphine occurred regardless of the effect of CCK-8 on unit firing. Thus, as has been found in anesthetized rats, CCK-8 appeared to enhance the inhibitory effects of a DA agonist on DA neurons.

Animals↗

Neuropeptide modulation of apomorphine-induced stereotyped behavior.

The nucleus accumbens contains many neuropeptides whose functions are presently unknown. The purpose of this study was to determine the extent to which these neuropeptides act in conjunction with the mesolimbic dopamine system. Microinjections of cholecystokinin, neurotensin, met-enkephalin, somatostatin, bombesin, as well as glutamate and muscimol, were made into the medial nucleus accumbens after systemic injection of apomorphine. Cholecystokinin and neurotensin, in nanogram doses, potentiated apomorphine-induced stereotypy. Met-enkephalin reduced, while somatostatin and bombesin were without effect on, apomorphine-induced stereotypy. In addition, both glutamate and muscimol potentiated this effect. These results suggest that several neuropeptides and amino acids act in the nucleus accumbens to modulate apomorphine-induced stereotyped behaviors.

Animals↗

Correlated asymmetries in striatal D1 and D2 binding: relationship to apomorphine-induced rotation.

Long-Evans derived rats were tested for nocturnal, amphetamine-induced and apomorphine-induced rotation (circling behavior); the rats' left and right striata were subsequently dissected and D1 and D2 receptor densities (Bmax) were assayed in the same striatal homogenates using [3H]SCH-23390 and [3H]N-methylspiperone, respectively. D1 and D2 Bmax values were correlated (r = 0.68). Moreover, left-right asymmetries in D1 and D2 Bmax values were more highly correlated (r = 0.84). Although asymmetries in D1 and D2 binding were not by themselves related to rotational behavior, an asymmetry in the ratio or balance of D1 and D2 binding was associated with the direction of apomorphine-induced rotation: the D1/D2 ratio of Bmax values was significantly higher in the striatum ipsilateral to the preferred direction of apomorphine-induced rotation. These results suggest that normal variations in numbers of D1 and D2 receptors are determined by a common mechanism, that D1 and D2 receptors are functionally coupled, and that, with respect to activation of striatal receptors, D1 is inhibitory and D2 is excitatory. The effects of apomorphine, a mixed D1 and D2 agonist, appear to reflect the balance between D1 and D2 receptors.

Animals↗

The effects of intracranial administration of the dopamine agonist apomorphine on penile reflexes and seminal emission in the rat.

Previous studies employing systemic administration of the dopamine agonist apomorphine have shown that the dose response curves for apomorphine's effects on penile reflexes and seminal emission differ, suggesting that experimentally separable populations of dopamine receptors regulate these two responses. The present experiments examined the locations of central nervous system DA receptors mediating genital responses in the restrained, supine rate by injecting apomorphine into the medial preoptic area and the paraventricular nucleus through chronic, indwelling cannulae. Medial preoptic area injections facilitated penile reflexes, but not seminal emission, while paraventricular injections facilitated seminal emission. These results suggest that systemically administered apomorphine may facilitate penile reflexes by acting on the medial preoptic area and may enhance seminal emission by acting on the paraventricular nucleus.

Animals↗

Neurotensin counteracts apomorphine-induced inhibition of dopamine release as studied by microdialysis in rat neostriatum.

Microdialysis in the neostriatum of the halothane-anesthetized male rats was used to study the effect of neurotensin on the release of dopamine and its metabolites in the absence or presence of systemic apomorphine treatment. Perfusate levels of dopamine, 3,4-dihydroxyphenylacetic acid (DOPAC) and homovanillic acid (HVA) were assayed by high-performance liquid chromatography in combination with electrochemical detection. Perfusion with neurotensin (1000 nM but not 10 nM) increased the dialysate levels of dopamine without affecting those of DOPAC and HVA. Systemic treatment with apomorphine (0.05 and 0.5 mg/kg, s.c.) reduced the dialysate levels of dopamine, DOPAC and HVA in a dose-related way. Neurotensin (10 nM but not 1 nM) counteracted the inhibitory effect of apomorphine on dialysate levels of dopamine without affecting those of DOPAC and HVA. The results indicate a facilitatory effect of neurotensin on dopamine release in rat neostriatum. It is suggested that activation of neurotensin receptors may cause a reduction in the affinity of dopamine autoreceptors, since the low dose of neurotensin is able to counteract the inhibitory effect of apomorphine on dopamine release.

3,4-Dihydroxyphenylacetic Acid↗

Apomorphine decreases extracellular GABA in the ventral pallidum of rats with 6-OHDA lesions in the nucleus accumbens.

Inhibition of a tonically active gamma-aminobutyric acid (GABA) projection from the nucleus accumbens to the ventral pallidum (VP) is thought to mediate the locomotor response elicited by dopamine in the nucleus accumbens. To evaluate this hypothesis, dopamine was depleted in the nucleus accumbens using 6-hydroxydopamine which produced an upregulated locomotor response to systemic apomorphine (0.2 mg/kg, s.c.). Simultaneously, the level of extracellular GABA in the VP was monitored using microdialysis. Apomorphine injection produced an elevation in locomotor activity only in the lesioned rats. While apomorphine reduced extracellular GABA in both control and lesioned rats, the reduction had an earlier onset and was more consistent in lesioned animals. Although the onset of the decline in extracellular GABA in the VP of lesioned rats corresponded to the onset of apomorphine-induced motor activity, a significant reduction in GABA persisted for 180 min, while the behavior returned to control levels by 60 min after injection. These data support a possible role for dopamine receptor-mediated inhibition of accumbal GABA neurons projecting to the VP in the initiation of locomotor activity.

Animals↗

3H-Apomorphine interactions with dopamine receptors in calf brain.

3H-apomorphine binds to membranes from areas of the corpus striatum and limbic system of calf brain saturable and with a drug specificity indicating that it labels dopamine receptors. In terms of drug specificity, log-logit displacement curve slopes and number of binding sites, 3H-apomorphine interacts with receptors in a manner more like 3H-dopamine than 3H-haloperidol. These properties of 3H-apomorphine binding are those of an apparently "pure" agonist in contrast to the partial agonist effects of apomorphine upon the dopamine-sensitive adenylate cyclase.

Animals↗

Selective labeling of apomorphine receptors by 3H-LSD.

There are at least two types of dopamine receptors: the 3H-dopamine or 3H-apomorphine receptor (with high or nM affinity for dopamine), and the 3H-neuroleptic receptor (with low or microM affinity for dopamine). While 3H-LSD can label the 3H-neuroleptic receptor, this study was done in order to label the 3H-apomorphine/dopamine receptor site. In the presence of excess phentolamine, serotonin and spiperone (to preclude binding to alpha-adrenergic, serotonergic and neuroleptic receptors, respectively) similar concentrations of dopaminergic drugs inhibited the binding (to calf caudate) of 3H-LSD and 3H-apomorphine. This is compatible with the concept that the 3H-apomorphine/dopamine receptor and the 3H-neuroleptic/dopamine receptor are separate.

Animals↗