Dopamine Overflow In Accumbens Research Articles

α3β4 nicotinic receptors in the medial habenula and substance P transmission in the interpeduncular nucleus modulate nicotine sensitization

The medial habenula-interpeduncular nucleus (MHb-IPN) pathway has recently been shown to modulate multiple effects nicotine in vivo, however it remains unclear which receptor subtypes in this pathway are critical for mediating these responses. To identify MHb and IPN receptors that play a role in nicotine reward, we studied receptors prevalent in these nuclei, including nicotinic acetylcholine receptors (nAChRs) and the receptor for substance P (neuokinin-1; NK1 receptor) using a model of behavioral and neurochemical sensitization to nicotine. Our results show that blockade of the α3β4 nAChR in the MHb, but not the IPN prevented increases in locomotor responding as well as increases in accumbal dopamine overflow in sensitized animals. Additionally, when NK1 receptors were blocked in the IPN, but not the MHb, a similar effect on sensitized responding was seen. Together, these results suggest that the MHb and IPN differentially modulate nicotine sensitization. Because the neurotransmission within these brain regions is primarily cholinergic and substance P ergic and these receptors are expressed in high density in both nuclei, these results could suggest a different neurophysiological signaling role or different neuroanatomical location of these receptors in this pathway. Furthermore, while α3β4 nAChRs have been suggested as a possible pharmacological target for nicotine addiction, this is the first evidence that substance P also plays a role in mediating responding to nicotine.

Adolescent nicotine sensitization and effects of nicotine on accumbal dopamine release in a rodent model of increased dopamine D2 receptor sensitivity

Our laboratory has reported neonatal quinpirole (D2/D3 agonist) treatment to rats increases dopamine D2 receptor sensitivity that persists throughout the animal's lifetime. This model appears to have clinical relevance to schizophrenia, and smoking is common in this population. Male and female Sprague-dawley rats were neonatally treated with quinpirole from postnatal (P) days 1–21. After habituation from P30 to 32, animals were administered saline or nicotine (0.3, 0.5, or 0.7mg/kg free base) every other day from P33 to 49 and locomotor activity was assessed. Generally, animals neonatally treated with quinpirole and administered nicotine during adolescence demonstrated increased behavioral activity and/or sensitization compared to animals neonatally given saline and sensitized to nicotine as well as controls. However, animals neonatally treated with quinpirole and given the 0.7mg/kg dose of nicotine demonstrated elevated activity throughout testing but did not show sensitization, and only mild sex differences were reported. Therefore, microdialysis was performed on male rats sensitized to the 0.5mg/kg dose of nicotine, and results revealed that neonatal quinpirole sensitized dopamine overflow in response to nicotine to 500% above animals neonatally given saline and sensitized to nicotine at peak levels. In addition, neonatal quinpirole increased the accumbal BDNF in response to nicotine compared to all other groups, and nicotine alone also produced significant increases in striatal and accumbal BDNF. This study reveals that neonatal quinpirole enhanced adolescent nicotine sensitization, accumbal dopamine overflow, and BDNF protein in response to nicotine, which may be related to changes in the brain's reward system.

Brain extracellular levels of the putative antipsychotic CI-1007 and its effects on striatal and nucleus accumbens dopamine overflow in the awake rat.

The compound CI-1007 (R-(+)-1,2,3,6-tetrahydro-4-phenyl-1 [(3-phenyl-3-cyclohexen-1-yl)methyl]pyridine maleate) has been identified as a partial dopamine agonist and putative antipsychotic in in-vitro and in-vivo neurochemical, neurophysiological and behavioural tests. By use of microdialysis in conjunction with high-performance liquid chromatography (HPLC) with electrochemical detection, the effects of the drug on brain dopamine release, previously observed in anaesthetized animals, were shown to occur in awake animals also. Detection of peripherally administered CI-1007 in the brain, i.e. evidence of the drug's ability to penetrate the blood-brain barrier, was achieved by use of in-vivo brain microdialysis in awake, freely moving rats and capillary HPLC in combination with tandem mass spectrometry (HPLC/MS/MS). Intravenous administration of CI-1007 (40 mg kg-1) significantly inhibits dopamine release in the nucleus accumbens, a region associated with dopamine hyperactivity in schizophrenia, while having a non-significant impact on the striatal dopamine neurotransmission which is critical to regular motor function. The differential neurochemical profile of the drug indicates its potential usefulness in treating positive disease symptoms and implies that its extrapyramidal side effects are lower than those of typical antipsychotics.

Effects of 18-methoxycoronaridine on ghrelin-induced increases in sucrose intake and accumbal dopamine overflow in female rats.

18-methoxycoronaridine (18-MC), a selective antagonist of α3β4 nicotinic receptors, has been previously shown, in rats, to reduce the self-administration of several drugs of abuse, reduce operant responding for sucrose, and prevent the development of sucrose-induced obesity. It has become increasingly apparent that there is a significant overlap between the systems regulating drug reward and food intake, therefore, we investigated whether 18-MC might modulate the effects of ghrelin, one of several orexigenic peptides recently implicated in both feeding and drug reward. In female Sprague-Dawley rats, we determined whether acute 18-MC treatment would reduce both ghrelin-induced increases in sucrose intake and ghrelin-elicited increases in accumbal dopamine levels. Pretreatment with 18-MC (20 mg/kg, i.p.), given prior to the administration of ghrelin (1 μg, lateral ventricle), blocked ghrelin-induced increases in sucrose (5%) intake in a two-bottle open access paradigm. Using in vivo microdialysis, 18-MC (both 20 and 40 mg/kg) prevented ghrelin (2 μg, intraventral tegmental area)-induced increases in extracellular dopamine in the nucleus accumbens. 18-MC had no effect on deposition of fat or on serum levels of glucose, triglycerides, and cholesterol in ghrelin-treated rats. The present results suggest that one potential mechanism by which 18-MC exerts its effects on palatable food consumption is via modulation of ghrelin's effects.

PRECLINICAL STUDY: Systemic administration of ghrelin induces conditioned place preference and stimulates accumbal dopamine

Ghrelin, preferably via hypothalamic circuits, is known to play a role in the control of energy balance, body weight homeostasis and appetite. In addition, it has recently been found that ghrelin injection into key structures of the mesolimbic dopamine (DA) system increases food intake in rodents and induces locomotor stimulation as well as accumbal DA overflow in mice. Thus, centrally administered ghrelin activates the mesolimbic DA system. However, ghrelin is mainly produced in and secreted from peripheral tissues. The present series of experiments were therefore undertaken to investigate the effects of peripherally injected ghrelin on mesolimbic DA system activity and, herewith, associated behaviours. It was found that intraperitoneal administration of ghrelin (0.33 mg/kg) induces locomotor stimulation, conditioned place preference and accumbal DA-overflow in mice. In conclusion, peripherally produced ghrelin, e.g. in the stomach, may activate the mesolimbic DA system. This suggests that the pre-prandial rise in plasma ghrelin may, via this system, increase the incentive value for motivated behaviours such as food seeking.

Presynaptic opioid and nicotinic receptor modulation of dopamine overflow in the nucleus accumbens.

Behaviorally relevant stimuli prompt midbrain dopamine (DA) neurons to switch from tonic to burst firing patterns. Similar shifts to burst activity are thought to contribute to the addictive effects of opiates and nicotine. The nucleus accumbens DA overflow produced by these drugs is a key element in their pathological effects. Using electrochemical techniques in brain slices, we explored the effects of opioids on single-spike and burst stimuli-evoked DA overflow in the dorsal and ventral striatum. In specific subregions of the nucleus accumbens, mu-opioids inhibit DA overflow elicited with single-spike stimuli while leaving that produced by burst stimuli unaffected. This is similar to published effects of nicotinic receptor blockade or desensitization, and is mediated by opioid receptor-induced inhibition of cholinergic interneurons. Whereas delta-opioids have similar effects, kappa-opioids inhibit evoked DA overflow throughout the striatum in a manner that is not overcome with high-frequency stimuli. These observations reveal remarkable mechanistic overlap between the effects of nicotine and opiates within the dopamine reward pathway.

Reduced psychostimulant effects on dopamine dynamics in the nucleus accumbens of μ-opioid receptor knockout mice

Dopamine neurotransmission in the nucleus accumbens plays a pivotal role in the reinforcing properties of drugs of abuse. Two interacting processes regulate nucleus accumbens dopamine overflow: release of dopamine from presynaptic terminals and the subsequent reuptake by dopamine transporters. Opioid neurotransmission, primarily through μ-opioid receptors has also been strongly implicated in drug reward. We have previously shown that mice lacking the μ-opioid receptor display decreased cocaine self-administration. In addition, we found decreased impulse activity of midbrain dopaminergic neurons and an increased GABAergic input to these neurons in μ-opioid receptor knockout mice. In the present study we investigated whether these changes in dopaminergic cell bodies are accompanied by altered dopamine dynamics at the terminal level. To that aim, we measured nucleus accumbens dopamine overflow using fast scan cyclic voltammetry. Our data demonstrate that in μ-opioid receptor knockout mice 1) the reuptake of dopamine in the nucleus accumbens is slower, and 2) the relative effect of cocaine and amphetamine on the reuptake of dopamine is smaller compared with wild type mice. These data provide a mechanism for the decreased reinforcing properties of cocaine observed in μ-opioid receptor knockout mice.

Is an α-conotoxin MII–sensitive mechanism involved in the neurochemical, stimulatory, and rewarding effects of ethanol?

Ethanol and nicotine are the most commonly abused drugs among human beings, and a large body of evidence, from both epidemiologic and preclinical studies, indicates that there is a positive correlation between intake of both drugs. Findings of studies from our research group have demonstrated that nicotinic acetylcholine receptors, especially those located in the ventral tegmental area, are important for the stimulatory, rewarding, and dopamine-enhancing effects of ethanol. Furthermore, results of recent work indicate that the α 4β 2 ∗ and the α 7 ∗ receptor subunits of the nicotinic acetylcholine receptors do not seem to be involved in the neurochemical and behavioral effects of ethanol in rodents. The aim of the current study was to investigate further the role of different nicotinic acetylcholine receptor subunits in the stimulatory, dopamine-enhancing, and rewarding effects of ethanol in rodents by using the peptide α-conotoxin MII (5 nmol; an antagonist of the α 3β 2 ∗, β 3 ∗, and α 6 ∗ subunits of the nicotinic acetylcholine receptor) administered locally into the ventral tegmental area. A significant reduction of ethanol-induced accumbal dopamine overflow, measured by means of in vivo microdialysis, and of locomotor stimulation was observed in mice. Furthermore, α-conotoxin MII was demonstrated to reduce voluntary ethanol intake significantly in both rats and mice. These results indicate that α-conotoxin MII–sensitive receptors may be important in mediating the stimulatory, dopamine-enhancing, and rewarding effects of ethanol, and that α-conotoxin MII–sensitive receptors may constitute targets for development of new adjuvant for treatment of ethanol dependence.

Oral sucrose stimulation increases accumbens dopamine in the rat.

Although taste can influence meal size and body weight, the neural substrate for these effects remains obscure. Dopamine, particularly in the nucleus accumbens, has been implicated in both natural and nonnatural rewards. To isolate the orosensory effects of taste from possible postingestive consequences, we investigated the quantitative relationship between sham feeding of sucrose and extracellular dopamine in the nucleus accumbens with microdialysis in rats. Sucrose intake linearly increased as a function of concentration (0.03 M, 18.07 +/- 2.41 ml; 0.1 M, 30.92 +/- 2.60 ml; 0.3 M, 43.28 +/- 2.88 ml). Sham feeding also stimulated accumbens dopamine overflow as a function of sucrose solution concentration (0.03 M, 120.76 +/- 2.6%; 0.1 M, 140.28 +/- 7.8%; 0.3 M, 146.27 +/- 5.05%). A second experiment used the same protocol but clamped the amount of sucrose ingested and revealed a similar, concentration-dependent dopamine activation in the nucleus accumbens. This is the first demonstration of a quantitative relationship between the concentration-dependent rewarding effect of orosensory stimulation by sucrose during eating and the overflow of dopamine in the nucleus accumbens. This finding provides new and strong support for accumbens dopamine in the rewarding effect of sucrose.

Behavioral sensitization to quinpirole is not associated with increased nucleus accumbens dopamine overflow

This study assessed the relationship between extracellular nucleus accumbens (NAc) dopamine (DA) concentrations and sensitized locomotor activation following repeated administration of the DA D2-like receptor agonist quinpirole. Locomotor activity measures and nucleus accumbens microdialysis samples were collected concurrently in response to the first (acute) and tenth (repeated) quinpirole injection (0.5 mg/kg s.c., every other day). Results indicate that acute quinpirole produced locomotor activation and that repeated quinpirole resulted in locomotor sensitization. Acute quinpirole significantly decreased the detection of extracellular concentrations of DA and the DA metabolites dihydroxyphenylacetic acid (DOPAC) and homovanillic acid (HVA) in the NAc. Following repeated quinpirole, basal NAc DA levels were decreased, whereas basal DOPAC levels were increased. Nevertheless, quinpirole challenge elicited a significant decrease in DA, DOPAC and HVA following repeated treatment. In addition, although acute quinpirole did not affect NAc levels of the serotonin metabolite 5-hydroxyindolacetic acid (5-HIAA), quinpirole challenge produced a significant increase in 5-HIAA levels following repeated treatment. Taken together, these data indicate that functional DA autoreceptor subsensitivity is not a necessary condition for the expression of behavioral sensitization to quinpirole. Instead, it appears that behavioral sensitization to quinpirole occurs predominantly as a consequence of neuroadaptations that are post-synaptic to DA release.

Role of different nicotinic acetylcholine receptors in mediating behavioral and neurochemical effects of ethanol in mice

Ethanol and nicotine are the most abused drugs, and it is well known that co-abuse of ethanol and nicotine is frequent in human beings. We have previously obtained results indicating that the ethanol-induced stimulation of both the mesolimbic dopamine system and locomotor activity may involve activation of central nicotinic acetylcholine receptors (nAChRs), especially those located in the ventral tegmental area. Different subpopulations of nAChRs have been identified, and, in the present series of experiments, we have studied the effects of various nAChR antagonists on the stimulation of dopamine overflow in the nucleus accumbens and on locomotor activity induced by ethanol in male mice. Ethanol (2.0 g/kg, i.p.) enhanced dopamine overflow in the nucleus accumbens by approximately 40%, measured by means of in vivo microdialysis in awake, freely moving mice. Mecamylamine (negative allosteric modulator of nAChR; 2.0 mg/kg, i.p.) blocked the ethanol-induced stimulation of both locomotor activity and accumbal dopamine overflow. Methyllycaconitine citrate (α 7 antagonist; 2.0 mg/kg, i.p.) and dihydro-β-erythroidine (competitive and selective α 4β 2 antagonist; 0.5 mg/kg, s.c.), in doses that had no marked effects per se, did not significantly reduce the behavioral and neurochemical stimulation caused by ethanol. The present results support the suggestion that the stimulatory effects of ethanol on locomotor activity and dopamine release do not involve the α 4β 2 or α 7 subunit compositions of the nAChR and that the effects of mecamylamine are mediated through a site not directly associated with the α 4β 2 or α 7 nAChR subunits.

Voluntary ethanol intake in the rat and the associated accumbal dopamine overflow are blocked by ventral tegmental mecamylamine

The mesocorticolimbic dopamine system is believed to be involved in mediating the positive reinforcing effects of drugs of abuse, including ethanol. The nicotinic acetylcholine receptor antagonist mecamylamine perfused via reversed microdialysis in the ventral tegmental area antagonizes the increase of accumbal extracellular dopamine levels after systemic ethanol, and, after systemic injection, lowers ethanol intake in the rat. In the present study the effect of ventral tegmental mecamylamine on ethanol intake and preference, as well as on extracellular accumbal dopamine levels, was investigated in the same animal. To this end, in vivo microdialysis using a double probe approach (one in the nucleus accumbens and one in the ventral tegmental area) was combined with an ethanol preference model invoking a free choice between a bottle of water and a bottle of ethanol 6% (v/v) solution. Wistar rats drinking more than 60% of their total daily fluid intake from the ethanol solution (ethanol high-preferring animals) were selected during a screening period and used for the experiments. The animals received vehicle or mecamylamine (100 μM) in the ventral tegmental area and were then presented with a choice between water and ethanol in a limited access paradigm to which they previously had been adapted. On the next day the rats that received vehicle day 1 now received mecamylamine, and vice versa. When treated with vehicle, ethanol intake and preference were unaltered, as compared to baseline behavior, and accumbal dopamine levels increased significantly to approximately 130% of the pre-drug baseline level. When receiving mecamylamine, ethanol intake and preference were reduced markedly and dopamine levels were unaltered, as compared to pre-drug baseline levels. The present results further indicate that nicotinic acetylcholine receptors in the ventral tegmental area are involved in the positive reinforcing effects of ethanol. Thus, mecamylamine or other antagonists specifically aimed at ventral tegmental nicotinic acetylcholine receptors could represent a new pharmacological treatment principle against alcohol abuse, the efficacy of which should be explored in high-scale alcohol consumers or alcoholics.

Nefazodone attenuates the behavioral and neurochemical effects of ethanol

This study evaluated the influence of nefazodone, a combined 5-HT 2A receptor antagonist and 5-HT reuptake inhibitor, on the behavioral and neurochemical effects of ethanol in nonselected male Wistar rats. In microdialysis experiments, ethanol (2.5 g/kg, IP) increased extracellular accumbal dopamine levels by 36% ( p = 0.0073) compared to baseline levels, and elevated the maximal DOPAC and HVA levels by 26% ( p = 0.0093) and 52% ( p = 0.0010), respectively. Nefazodone (50 mg/kg, SC) per se increased accumbal dopamine levels by 28% ( p = 0.0199), but, when injected 40 min before ethanol, reduced the ethanol-induced elevation of accumbal dopamine overflow ( p = 0.0132) and decreased the ethanol-induced HVA levels ( p = 0.0159). In an ethanol(6% v/v)/water free-choice paradigm, nefazodone (50 mg/kg, SC) decreased ethanol intake by 51% ( p = 0.0251) and preference by 22% ( p = 0.0251) in high- but not low-preferring rats from a nonselected Wistar strain. These results show that nefazodone modulates the mesolimbic dopamine system in a dopamine activity-dependent manner, and influences the neurochemical and behavioral effects of ethanol in the rat.

Nicotine and heroin augment cocaine-induced dopamine overflow in nucleus accumbens

The current public debate on nicotine concentrates on the abuse potential of nicotine per se. However, little is known about the interaction of nicotine with other drugs of well-established abuse liability such as cocaine. Indeed, cigarette smoking increases the intake of cocaine and other drugs of abuse. In order to test if these epidemiological data are reflected in a neurochemical correlate of the reinforcing effects of drugs of abuse, i.e., dopamine overflow in the nucleus accumbens, in vivo brain microdialysis was used to examine the effects of nicotine and cocaine either alone or in combination in freely moving rats. Furthermore, the effects of the nicotine+cocaine combination were compared to another drug combination of high abuse potential, i.e., heroin+cocaine (`speedball'). Both nicotine+cocaine as well as heroin+cocaine stimulated nucleus accumbens dopamine overflow in an additive manner. Repeated intermittent administration of nicotine did not significantly alter the effects of a subsequent challenge with the nicotine+cocaine combination. These data suggest that the clinical-epidemiological findings on either drug combination are reflected in a stimulatory interaction on nucleus accumbens dopamine overflow that is additive. No significant tolerance seems to develop to this effect of nicotine. These neurochemical findings support behavioral data suggesting that the reinforcing effects of cocaine and heroin are additive and predict that nicotine will enhance the reinforcing effects of cocaine.

Accumbal dopamine overflow after ethanol: Localization of the antagonizing effect of mecamylamine

It has been suggested that ethanol exerts its mesolimbic dopamine activating effects and its reinforcing effects via interaction with central nicotinic acetylcholine receptors, thus providing a basis for the often observed covariation between ethanol and nicotine consumption. We have previously demonstrated that the central nicotinic acetylcholine receptor antagonist mecamylamine totally counteracts the ethanol-induced elevation of extracellular dopamine in the nucleus accumbens, as measured by in vivo microdialysis. A contribution of peripheral nicotinic receptor blockade could, however, not be excluded. In the present study, mecamylamine (1.0 mg/kg, i.p.) again totally counteracted the ethanol-induced dopamine overflow, as measured by in vivo microdialysis, while the quarternary nicotinic receptor antagonist hexamethonium (10 mg/kg, i.p.) did not. Furthermore, the increase in accumbal dopamine overflow after systemic ethanol (2.5 g/kg, i.p.) was counteracted by local perfusion of mecamylamine (50 μM) in the ipsilateral ventral tegmental area, but not by mecamylamine perfusion in the nucleus accumbens. Ethanol-induced accumbal dopamine overflow was also counteracted by perfusion of hexamethonium (250 μM) in the ventral tegmental area. These results provide further evidence that ethanol-induced activation of the mesolimbic dopamine system is mediated via stimulation of central nicotinic acetylcholine receptors, and that the receptor population within the ventral tegmental area may be the most important in this regard. It is suggested that antagonists of central nicotinic acetylcholine receptors may be useful in the treatment of alcoholism.

Effect of prior ethanol experience on dopamine overflow in accumbens of AA and ANA rats

The purpose of this study was to investigate the effect of repeated ethanol administration on dopamine overflow in the nucleus accumbens of alcohol-preferring AA (Alko Alcohol) and alcohol-avoiding ANA (Alko Nonalcohol) rats. Dopamine is a possible mediator of the reinforcing effects of ethanol, but it has previously been shown that ethanol-naïve alcohol-preferring AA and alcohol-nonpreferring ANA rats do not differ in their dopaminergic reaction to an intraperitoneal ethanol injection (0.5–2.0 g/kg), as assessed by measuring extracellular dopamine in the nucleus accumbens with in vivo microdialysis. Here a group of AA rats drank 10% (v/v) ethanol voluntarily — continual access for 5–15 days, limited access for 3 weeks — while a yoked group of AA rats and a yoked group of ANA rats received the same amount intragastrically by intubation. The rats were implanted with guide cannulas on the fourth week of limited access. Dopamine overflow was monitored in the microdialysis perfusate after 1 g/kg i.p. ethanol. The AA and the ANA rats that received ethanol non-contingently showed the same dopaminergic response to this as naïve animals have before. The group that had ingested the ethanol voluntarily showed, however, a significantly smaller increase in dopamine after 1 g/kg ethanol i.p. This suggests that the active behavior associated with obtaining the contingent drug may have an important impact on the reactions of the dopamine system to the drug, producing different results than when the same drug is admistered by other routes. The hypothesis that dopamine mediates ethanol reinforcement in AA rats is not supported by the results.

Transmitter release from transplants of fetal ventral mesencephalon or locus coeruleus in the rat frontal cortex and nucleus accumbens: effects of pharmacological and behaviorally activating stimuli

The present study was performed in order to establish whether dopamine (DA) release from behaviorally functional intracerebral DA transplants is dependent on changes in neuronal impulse flow, and is under control of the host brain. Rats were subjected to combined intraventricular and ventral tegmental injections of 6-hydroxydopamine (6-OHDA) in order to obtain a severe bilateral lesion of the ascending mesocorticolimbic DA projections. Cell suspension grafts of fetal ventral mesencephalic neurons were thereafter implanted into the medial frontal cortex (MFC) and the nucleus accumbens (NAc). Since the neurotoxin injections removed also the ascending noradrenergic systems, fetal locus coeruleus neurons were added to the graft suspension in one group of animals. Age-matched lesion-only and normal animals served as controls. The lesion-induced alterations in spontaneous, amphetamine- and apomorphine-induced locomotor activity and in a skilled paw reaching task were evaluated before transplantation, and at 3 and 6 months post-grafting. Microdialysis probes were finally implanted in the MFC and NAc in order to monitor extracellular DA and noradrenaline (NA) levels ( i) during administration of pharmacological agents which augment or depress catecholamine release in the intact brain; ( ii) during exposure of the rats to stressful manipulations ( handling and immobilization) or appetitive stimuli ( eating) known to enhance cortical and limbic DA or NA release in intact animals. The lesion-induced reduction in amphetamine-induced locomotor activity was reversed in all grafted animals, which also showed a higher than normal spontaneous overnight activity. Daytime spontaneous locomotor activity (which was reduced in the lesion-only rats) as well as apomorphine-induced hyperactivity was reversed by the grafts of DA neurons only. By contrast, the lesion-induced impairment in skilled forelimb use was not alleviated by the grafts. The grafted DA neurons restored normal steady-state DA overflow in the NAc, whereas they enhanced cortical DA overflow to significantly higher than normal levels. Restoration of both cortical and striatal NA overflow was observed in the group that received mixed DA and NA grafts, whereas animals that received DA grafts only did not differ from the lesioned controls. The changes in extracellular DA and NA levels measured in the grafted MFC and NAc under potassium depolarization (100 mM KCl), inhibition of terminal catecholamine reuptake (10 μM nomifensine), and sodium channel blockade (1 μM TTX) indicated that graft-derived DA or NA release had normal neuronal properties, and was dependent on an intact axonal impulse flow. The grafted DA and NA neurons lacked the ability to respond to immobilization stress with enhanced transmitter release. However, most of the animals that received DA grafts exhibited an enhanced DA overflow in the NAc following handling stimulation. When the animals were engaged in eating, a significant 20% increase in DA overflow was detected in the grafted cortices, and a marginally significant 58% increase in DA overflow was measured in the DA-grafted NAc. By contrast, neither cortical nor accumbens DA overflow was significantly affected during eating behavior in the lesion-only controls. NA overflow from the grafted locus coeruleus neurons was also unaffected by the appetitive stimulus. The present results indicate that DA or NA release from intracortically and intrastriatally grafted DA and NA neurons is dependent on changes in axonal impulse flow. The findings of enhanced DA release from intracortically grafted DA neurons during eating and from intrastriatally grafted DA neurons during rewarded behavior and handling stimulation, suggest that the transplanted DA neurons are reached by physiologically relevant inputs from the host brain. On the other hand, the lack of response to forced immobilization (i.e. a stimulus which is normally a powerful activator of cortical DA release), reveals that the functional integration of the grafted DA neurons with the host brain is incomplete.