Interactions between morphine and the morphine-glucuronides measured by conditioned place preference and locomotor activity

After intake of heroin or morphine, active metabolites are formed in the body. The two most important morphine metabolites are morphine-6-glucuronide (M6G) and morphine-3-glucuronide (M3G). M6G and M3G are present for longer time periods and in higher concentrations than the parent drug, but their potential contribution to reward and to development of dependence and addiction is not clear.We tested the effects of morphine and M6G separately (doses of 10, 20, 30 and 50 µmol/kg), administered together, and also in combination with with 200 µml/kg M3G in male C57BL/6J-Bom mice. M3G in doses of 50, 100, 200, 300 and 400 µmol/kg were also tested alone. We evaluated the rewarding effects in a conditioning place preference (CPP) model and the psychomotor stimulating effects by recording locomotor activity.Mice were subjected to three consecutive conditioning days with drugs or saline before testing. Changes in locomotor activity from conditioning day one to day three were also compared to the expression of CPP on the test day.This study revealed that coadministration of morphine and M6G induced CPP of similar magnitude to the sum of equimolar doses of these compounds alone, and different ratios of the two drugs did not affect the results. M3G did not cause CPP and reduced the CPP induced by both morphine and M6G when coadministered with these drugs. Morphine induced locomotor activity was reduced by coadministration of M3G, but this was not seen when M3G was co-injected with M6G. The changes in locomotor activity during the conditioning periods did not correlated with the expression of CPP.This study revealed that the morphine-glucuronides in different and complex ways can influence the pharmacological effects of psychomotor activation and reward observed after intake of morphine.     

Conditioned place preference induced by morphine and morphine-6-glucuronide in mice

Morphine-6-glucuronide (M6G), an active metabolite of morphine has been shown to produce analgesia and fewer side effects than morphine, and the introduction of M6G as a new drug for treatment of postoperative pain is planned in 2007. Following morphine intake in humans, the metabolites morphine-3-glucuronide (M3G) and M6G are present in substantial concentrations and for longer periods than the parent drug. The possible reward effects of the morphine glucuronides have previously not been well studied. In the present study, conditioned place preference (CPP) was recorded after conditioning with subcutaneous injections of 5, 10, 20, 30 or 50 micromol/kg morphine or M6G, or 240 or 500 micromol/kg M3G in C57BL/6J-Bom mice, using a biased two compartment ("closed" and "open") counterbalanced paradigm. CPP was induced after treatment with both morphine and M6G with dose dependent increase up to 30 micromol/kg after treatment in the "closed" compartment. No dose response was observed in the "open" compartment, with maximal CPP after 10 micromol/kg morphine or M6G. M3G caused a tendency of condition place aversion (CPA), although not statistically significant. In the present study morphine and M6G demonstrated comparable reward effects, at doses that differed depending on which compartment the mice were conditioned in. M3G showed a tendency to exhibit aversive properties.     

The role of mu opioid receptors in psychomotor stimulation and conditioned place preference induced by morphine-6-glucuronide

Previous studies have shown that morphine-6-glucuronide (M6G), a metabolite of morphine, induces reward and psychomotor stimulation but the role of the mu opioid receptor in these actions of the drug is not fully characterized. Thus, using mice lacking exon-2 of the mu opioid receptor and their wild-type littermates/controls, we determined the role of this receptor in psychomotor stimulation, sensitization, and conditioned place preference (CPP) induced by M6G. For comparison, we also assessed the role of the mu opioid receptor in the rewarding action of morphine. For the measurement of locomotor activity and sensitization, mice were habituated to motor activity chambers for 1h, then injected with M6G (10mg/kg) and locomotor activity was recorded for an additional 1h. The same treatment was given for five days and mice were tested for sensitization a week later. For the CPP experiments, mice were tested for baseline place preference on day 1, then received single or repeated alternate-day saline/drug or drug/saline conditioning and tested for CPP the following day. Mice were also tested for CPP under a drugged state. M6G induced psychomotor stimulation, a response that was enhanced upon repeated administration of the drug, showing that locomotor sensitization developed to the motor stimulatory action of M6G. However, M6G induced a weaker CPP response compared to morphine. None of these actions of M6G was detected in mice lacking the mu opioid receptor. Together, the current results suggest that M6G induces psychomotor stimulation and a weaker rewarding action via the mu opioid receptor.     

Morphine-3-glucuronide inhibits morphine induced, but enhances morphine-6-glucuronide induced locomotor activity in mice

The main metabolite of morphine, morphine-3-glucuronide (M3G) has no opioid effects. Some studies have rather indicated that it antagonizes the antinociceptive and respiratory depressive effects of both morphine and the active metabolite morphine-6-glucuronide (M6G). We studied the possible influence of M3G on the psychostimulant properties of morphine and M6G measured by locomotor activity. Mice were given two injections, one with either 80, 240 or 500 micromol/kg M3G or saline followed by an injection of 20 or 30 micromol/kg morphine or M6G. M3G influenced the locomotor activity induced by both morphine and M6G, but in opposite directions. M3G reduced the morphine induced locomotor activity during the first hour following morphine injection in a concentration dependent manner. M3G pretreatment did not significantly influence brain concentrations of morphine indicating that the interaction was of a pharmacodynamic type. In contrast M3G pretreatment increased the M6G induced locomotor activity. M3G pretreatment increased serum and brain M6G concentrations to an extent indicating that this interaction was mainly of a pharmacokinetic type. In conclusion our results disclose complicated interactions between morphine and its two metabolites with respect to induction of locomotor activity and possibly also with respect to mechanisms related to drug reward.     

Effects of beta-funaltrexamine and naloxonazine on single-trial morphine-conditioned place preference and locomotor activity

The current study assessed the ability of the selective irreversible mu-opioid receptor antagonists beta-funaltrexamine (betaFNA) and naloxonazine (NALZ) to alter the locomotor and rewarding effects of a single intravenous injection of morphine using the conditioned place preference (CPP) model. In the first experiment, rats were conditioned with a single injection of morphine (10 mg/kg iv) paired with one compartment of a CPP apparatus and then were tested for CPP at either 1 or 7 days after conditioning. Rats showed hypoactivity following acute morphine on the conditioning trial and showed CPP when tested either 1 or 7 days later. In the next experiments, rats were pretreated with betaFNA (20 mg/kg sc, 20 h before conditioning), NALZ (15 or 30 mg/kg sc, 24 h before conditioning) or saline and then were conditioned with a single injection of morphine (10 mg/kg iv) or saline. Pretreatment with NALZ alone, but not betaFNA, significantly decreased locomotor activity; neither antagonist alone produced a significant shift in preference for either compartment of the CPP apparatus. Pretreatment with either betaFNA or NALZ blocked completely morphine-induced hypoactivity, but neither antagonist had a significant effect on morphine CPP. These results indicate that mu-opioid receptors are more critically involved in acute morphine-induced hypoactivity than in acute morphine reward.     

Behavioural sensitization in mice induced by morphine-glucuronide metabolites

Sensitization is thought to be involved in central aspects of drug addiction. Both morphine-3-glucuronid (M3G) and morphine-6-glucuronid (M6G) are rapidly formed in high concentrations shortly after heroin and morphine consumption. Their role in the development of sensitization has not previously been studied. In our study, mice received three injections of M6G or morphine at six day intervals. M6G induced locomotor sensitization comparable to morphine as early as the first injection. In a second experiment two injections of M6G or morphine were given, separated by 6, 12, 18, 24 or 30 days. A sensitized response was observed for both morphine and M6G up to 18 days after the first injection. In a third experiment with two injections, the first with M6G and the second with morphine, or the opposite sequence, M6G did not induce cross-sensitization to morphine although morphine induced cross-sensitization to M6G. Finally, pretreatment with M3G induced sensitization of morphine locomotor activity but not M6G. In conclusion M6G induced long-lasting sensitization similar but not identical to morphine. M3G was shown to sensitize morphine induced locomotor activity in a similar way to morphine pretreatment. This suggests that morphine-glucuronide metabolites may play a role in the development of addiction to morphine.     

Region-specific effects of brain corticotropin-releasing factor receptor type 1 blockade on footshock-stress- or drug-priming-induced reinstatement of morphine conditioned place preference in rats

Rationale Systemic injections of the selective corticotropin-releasing factor 1 (CRF1) receptor antagonist CP-154,526 attenuate footshock-stress-induced reinstatement of heroin and cocaine seeking and morphine conditioned place preference (CPP). Intracranial injections of the nonselective CRF receptor antagonist d-Phe-CRF into the bed nucleus of the stria terminalis (BNST), but not the amygdala, attenuate footshock-induced reinstatement of cocaine seeking. However, the brain sites involved in the effect of CP-154,526 on footshock-induced reinstatement of opiate seeking are unknown. Objective We used a CPP version of the reinstatement model to examine the role of CRF1 receptors in the BNST, amygdala, and nucleus accumbens (NAc) in footshock- or drug-priming-induced reinstatement of extinguished morphine CPP. Methods Rats acquired morphine CPP over a period of 8 days during which they were given four morphine (10 mg/kg s.c.) and four saline injections and were subsequently confined to distinct chambers for 50 min. Subsequently, the morphine CPP was extinguished in 14 daily sessions during which rats were given saline injections and given access to both the saline- and morphine-paired chambers. The rats were then tested for reinstatement of morphine CPP induced by priming injections of morphine (0 or 3.0 mg/kg s.c.) or by exposure to intermittent footshock (15 min, 0.5 mA). Prior to the test sessions, the rats were given intracranial injections of CP-154,526 (1.0 μg) or vehicle into the BNST, amygdala, or NAc. Results CP-154,526 injections into the BNST, but not the amygdala or NAc, attenuated footshock-stress-induced reinstatement of morphine CPP. In contrast, CP-154,526 injections into the amygdala or NAc, but not the BNST, attenuated morphine-priming-induced reinstatement of morphine CPP. Conclusion The present results demonstrate dissociable roles of CRF1 receptors in the BNST, amygdala, and NAc in footshock-stress- vs morphine-priming-induced reinstatement of drug CPP. The authors J. Wang and Q. Fang contributed equally to this work.     

下边缘皮质中神经元周围基质网络（PNNs）的降解联合消退训练能够抑制吗啡奖赏记忆的复燃

目的：利用硫酸软骨素酶（ChABC）降解下边缘皮质神经元周围基质网络（PNNs）之后,观察对于吗啡奖赏记忆复燃的影响。方法：采用条件性位置偏爱（CPP）模型,用核团微注射方法。结果：（1）与PBS对照组相比,消退训练前在下边缘皮质微注射ChABC将PNNs降解之后联合消退训练能够抑制消退后低剂量吗啡引起的CPP复燃;（2）与对照组相比,吗啡CPP训练前下边缘皮质微注射ChABC,对于吗啡CPP的获得没有影响。结论：本研究发现下边缘皮质神经元周围基质网络（PNNs）/硫酸软骨素蛋白聚糖（CSPGs）降解之后,可以抑制消退后低剂量药物引起的觅药行为的复现。这种作用可能是通过PNNs/CSPGs降解能够增强吗啡奖赏消退记忆来发挥作用的。     

Further studies on nicotine-induced conditioned place preference in the rat

Rats received subcutaneous (SC) injections of either nicotine (NIC, 0.001 to 2.0 mg/kg) or saline (SAL, 1 ml/kg) immediately prior to conditioning sessions in a conditioned place preference (CPP) paradigm. NIC was paired for 3 conditioning sessions with one environment of a 3 compartment CPP apparatus; SAL was paired with another environment. The animals were then tested for place preference by determining the proportion of time spent in each compartment during a 15 min test session. A dose-response curve was obtained for the place conditioning effect of nicotine as measured by its ability to alter baseline preferences calculated from control rats. NIC's place preference, but not place aversion, effect was linearly correlated with respect to dosage within the range of 0.1 to 0.8 mg/kg. NIC, 0.8 mg/kg, induced a place preference when it was administered immediately prior to conditioning sessions, but not when administered 20, 60 or 120 min prior to the sessions. Three repeated conditioning and testing cycles, or the daily administration of NIC for 2 weeks between conditioning and testing cycles had little or no effect on NIC place conditioning. Lobeline (2, 10 and 20 mg/kg) or cotinine (1 to 50 mg/kg) failed to condition a place preference. NIC, 0.1 or 1.2 mg/kg SC, administered to rat pups on postnatal days 5 through 8, did not alter subsequent place preference (induced by 0.8 mg/kg of NIC) measured at approximately 40 and 70 days of age. Periodic measurements of spontaneous motor activity, forelimb grip strength and negative geotaxis were unaltered by the perinatal exposure to nicotine.     

Conditioned rewarding effects of morphine and methadone in mice pre-exposed to cocaine

BackgroundMethadone is widely accepted as the most effective treatment of opioid dependence. However, clinical observations indicate that the medication is less effective in individuals abusing cocaine. Diminished therapeutic efficacy of methadone in cocaine users is intriguing, but its mechanism has not been studied.MethodsHere, the conditioned place preference (CPP) procedure was used to examine the effects of the dose, number of conditioning sessions and pre-exposure to cocaine on the rewarding effects of morphine and methadone. Vehicle-pre-exposed and cocainesensitized mice (five injections of 10 mg/kg over 16 days) were conditioned using methadone (0, 0.1, 0.5, 3, and 5 mg/kg) or morphine (0, 1, and 10 mg/kg). Place preference was measured after one and again after two additional conditioning sessions.ResultsAs expected, morphine at 10 mg/kg produced CPP following just one conditioning session. While a single conditioning session with 1 mg/kg of morphine produced no CPP, the rewarding effect became apparent following two additional conditioning sessions as well as in mice pre-exposed to cocaine. Methadone produced CPP following one conditioning session at doses of 0.5, 3 and 5 mg/kg. However, unlike with morphine, methadone's rewarding effect was not enhanced by two additional conditioning sessions or by pre-exposure with cocaine.ConclusionsPrior exposure to cocaine increases unconditioned motivational effects of morphine but not of methadone.     

下边缘皮质中神经元周围基质网络(PNNs)的降解联合消退训练能够抑制吗啡奖赏记忆的复燃

目的:利用硫酸软骨素酶(ChABC)降解下边缘皮质神经元周围基质网络(PNNs)之后,观察对于吗啡奖赏记忆复燃的影响。方法:采用条件性位置偏爱(CPP)模型,用核团微注射方法。结果:(1)与PBS对照组相比,消退训练前在下边缘皮质微注射ChABC将PNNs降解之后联合消退训练能够抑制消退后低剂量吗啡引起的CPP复燃;(2)与对照组相比,吗啡CPP训练前下边缘皮质微注射ChABC,对于吗啡CPP的获得没有影响。结论:本研究发现下边缘皮质神经元周围基质网络(PNNs)/硫酸软骨素蛋白聚糖(CSPGs)降解之后,可以抑制消退后低剂量药物引起的觅药行为的复现。这种作用可能是通过PNNs/CSPGs降解能够增强吗啡奖赏消退记忆来发挥作用的。     

Acquisition and expression of ethanol-induced conditioned place preference in mice is inhibited by naloxone

The effects of opioid antagonists on conditioned reward produced by ethanol provide variable and sometimes conflicting results, especially in mice. In the present set of experiments, male C57BL/6 mice received 4 vehicle and 4 ethanol conditionings, and the rewarding effects of ethanol were assessed in an unbiased version of the conditioned place preference (CPP) apparatus and an unbiased stimulus assignment procedure. Intraperitoneal (ip) administration of ethanol (2 g/kg, but not 1 g/kg) resulted in the conditioned reward when conditionings lasted for 6 min but not when conditioning lasted for 20 min. Administration of the non-selective opioid receptor antagonist naloxone (1 and 5 mg/kg) before the conditionings attenuated the acquisition of ethanol-induced place preference. Naloxone (1 mg/kg) also inhibited expression of the CPP response, but it did not alter the preference of vehicle-conditioned mice, suggesting the lack of its own motivational effects in this experimental setting. Taken together, the present results suggest that an unbiased version of ethanol-induced CPP in C57BL/6 mice could be a valid model for the study of the motivational effects of ethanol, confirming and expanding previous findings that have demonstrated inhibitory effects of opioid receptor antagonist on alcohol conditioned reward.     

Cross-reinstatement by cocaine and amphetamine of morphine-induced place preference in mice

The cross-reinstatement by psychostimulants of a conditioned place preference (CPP) induced by morphine was evaluated in mice. In Experiment 1, we examined the effects of a single dose of cocaine and amphetamine on a previously extinguished morphine CPP. After acquisition of CPP induced by morphine (40 mg/kg), animals underwent daily extinction sessions of 15 min duration until the CPP was extinguished. Subsequently, animals received a non-contingent injection of cocaine (25 mg/kg) or amphetamine (4 mg/kg), which produced the reinstatement of the extinguished morphine-induced CPP. In Experiment 2, we evaluated the reinstating effects of several priming doses of cocaine (Experiment 2A) or amphetamine (Experiment 2B). As in the first experiment, after conditioning with morphine (40 mg/kg), mice underwent daily 15 min extinction sessions. When the preference was no longer evident, we tested the effects of cocaine (0, 6.25, 12.5, 25 and 50 mg/kg) and amphetamine (0, 0.5, 1, 2 and 4 mg/kg) on the reinstatement of CPP. Doses from 12.5 mg/kg of cocaine upward and doses from 1 mg/kg of amphetamine upward effectively reinstated CPP. Our results demonstrate cross-reinstatement with psychostimulants and opiates, suggesting that in abstinent individuals, drug exposure can produce craving for the previously abused drug and relapse.     

Prenatal blockade of androgen receptors reduces the number of intromissions needed to induce conditioned place preference after paced mating in female rats

Estrous female rats pace their coital contacts to control the rate of cervical/vaginal stimulation. Females that receive at least 10 paced intromissions develop a reward state, evaluated by conditioned place preference (CPP), whereas females that do not pace their coital contacts do not develop CPP. We asked if the blockade of androgen receptors could modify the number of intromissions needed during paced mating to develop CPP. Pregnant females received daily injections of the androgen receptor antagonist flutamide from day 12 of pregnancy until pups were born. When adults, females exposed prenatally to flutamide or vehicle were ovariectomized and hormonally primed to evaluate if paced and non-paced mating induced CPP. The prenatal flutamide treatment did not affect the capacity of females to develop CPP to a systemic morphine injection. Flutamide-treated females that paced their sexual contacts developed CPP with fewer intromissions than control females. No effect on conditioning was observed when females were not allowed to pace their sexual contacts. The results suggest the existence of a threshold of cervical/vaginal stimulation that correlates with the induction of a reward state and that this threshold can be reduced by prenatal blockade of androgen receptors.     

Conditioned place preference: no tolerance to the rewarding properties of morphine

The effect of repeated morphine administration on conditioned place preference (CPP) using a novel treatment schedule, i.e., drug treatment was always contingent with the conditioned environmental stimuli, was investigated. We also examined whether changes in the μ- and κ-opioid receptor binding occurred in the brain of morphine-treated animals. Intraperitoneal (i.p.) administration of morphine (2 and 10 mg/kg) induced a place preference after 8 daily conditioning trials (4 morphine injections on alternate trials), the level of preference being the same with the two doses of the opiate. No change in place preference was observed in the morphine-treated rats at 2 mg/kg, when animals were further trained up to a total of 32 conditioning trials (16 morphine injections). Conversely, after 20 conditioning trials (10 morphine injections), a stronger CPP response developed in the morphine-treated rats at 10 mg/kg. Signs of morphine withdrawal were never detected in morphine-treated rats during the experiment. Loss of body weight (index of opiate dependence) was not observed either 24 h or 48 h after the last morphine administration. μ- and κ-opioid receptor density and affinity were not affected by repeated morphine administrations at either dose. The results demonstrate that no tolerance develops to the rewarding properties of morphine. Indeed, a sensitisation effect may occur at increasing doses of the opiate. Furthermore, changes in the rewarding effect of morphine are not dependent upon alterations in opioid receptors involved in the reinforcing mechanisms. Received: 31 October 1996 / Accepted: 7 February 1997     

Effects of unilatral‐ and bilateral inhibition of rostral ventral tegmental area and central nucleus of amygdala on morphine‐induced place conditioning in male Wistar rat

The rostral ventral tegmental area (VTAR) and central nucleus of amygdala (CeA) are considered the main regions for induction of psychological dependence on abused drugs, such as morphine. The main aim of this study was to investigate the transient inhibition of each right and left side as well as both sides of the VTAR and the CeA by lidocaine (2%) on morphine reward properties using the conditioned place preference (CPP) method. Male Wistar rats (250±20 g) 7 days after recovery from surgery and cannulation were conditioned to morphine (7.5 mg/kg) in CPP apparatus. Five minutes before morphine injection in conditioning phase, lidocaine was administered either uni‐ or bilaterally into the VTAR (0.25 μL/site) or CeA (0.5 μL/site). The results revealed that lidocaine administration into the left side, but not the right side of the VTAR and the CeA reduced morphine CPP significantly. The reduction was potentiated when lidocaine was injected into both sides of the VTAR and the CeA. The number of compartment crossings was reduced when lidocaine was injected into both sides of the VTAR and the CeA as well as the left side. Rearing was reduced when lidocaine was injected into the right, but not the left side of the VTAR. Sniffing and rearing increased when animals received lidocaine in the right side and reduced in the group that received lidocaine in the left side of the CeA. It was concluded that the right and the left side of VTAR and the CeA play different roles in morphine‐induced activity and reward.     

Intra-hippocampal inhibition of protein kinase AII attenuates morphine-induced conditioned place preference

Morphine and other drugs of abuse modulate protein kinase A (PKA) signaling within the mesolimbic reward pathway. Using a balanced conditioned place preference (CPP) paradigm, we studied the possible involvement of protein kinase AII (PKA II) on the acquisition, expression and consolidation of morphine place conditioning in male Wistar rats. Subcutaneous administration of various doses of morphine sulfate (1-9 mg/kg) induced CPP in a dose-dependent manner. H-89, a selective PKA II inhibitor, was administered into CA1 region of the hippocampus at 1, 2.5 and 5 microM/rat. Using a 3-day schedule of conditioning, it was found that the H-89 did not produce a significant place preference or place aversion. H-89 (1, 2.5 and 5 microM/rat) significantly reduced the time spent by rats in the morphine compartment when given immediately after each conditioning session (consolidation), whereas it had no effect when administered before morphine during the conditioning phase (acquisition) or before testing for place preference in the absence of morphine (expression). It is concluded that the PKA II may play an active role in the consolidation of reward-related memory of morphine in CA1 region of the hippocampus.     

Repeated administration of dopaminergic agents in the dorsal hippocampus and morphine-induced place preference

The aim of the present experiments was to investigate whether repeated intra-hippocampal CA1 (intra-CA1) administration of dopaminergic agents can affect morphine-induced conditioned place preference (CPP). Effects of repeated intra-CA1 injections of dopamine (DA) receptor agonists and antagonists on morphine-induced CPP in rats were investigated using an unbiased 3-day schedule of place conditioning. Animals receiving once-daily subcutaneous (s.c.) injections of morphine (1-9 mg/kg) or saline (1.0 ml/kg, s.c.) showed a significant place preference in a dose-dependent manner: the maximum response was observed with 3 mg/kg morphine. Three days' intra-CA1 injections of apomorphine (0.25-1 microg/rat) followed by 5 days free of the drug, significantly decreased morphine CPP (1 and 3 mg/kg, s.c.). Moreover, pre-treatment with the highest dose of apomorphine (1 microg/rat) altered the effect of morphine to an aversive response. The morphine (1 and 3 mg/kg) CPP was also significantly decreased in animals that previously received three intra-CA1 injections of SKF 38393 (2-9 microg/rat), quinpirole (1-3 microg/rat) or sulpiride (1-3 microg/rat), and significantly increased in animals that had previously received three intra-CA1 injections of SCH 23390 (0.02 microg/rat). The 3-day pre-treatment with apomorphine, SKF 38393 or quinpirole reduced locomotor activity in the test session, while SCH 23390 and sulpiride did not have any influence on locomotor activity. It is concluded that repeated injections of DA receptor agents in the dorsal hippocampus, followed by 5 days free of the drugs, can affect morphine reward.     

Nalbuphine is effective in decreasing the rewarding effect induced by morphine in rats

Nalbuphine, a kappa-opioid agonist and mu-opioid partial agonist, has been widely used as an analgesic or an adjuvant with morphine in clinics. In rats, it attenuates tolerance and physical dependence caused by morphine, when co-administered. In this study, we investigated the effect of nalbuphine on morphine reward. Using the conditioned place preference (CPP) paradigm in rats, we demonstrated that co-administration of nalbuphine (1mg/kg, i.p.) with morphine (5mg/kg, i.p.) during conditioning could completely block the CPP induced by morphine. However, in experiments examining locomotor activity in rats, nalbuphine showed no effect on the development of behavioral sensitization induced by reported morphine administration. In microdialysis experiments, morphine induced a significant increase in the dopamine metabolites 3,4-dihydroxyphenylacetic acid and homovanillic acid in the shell region of the nucleus accumbens. Co-administration of nalbuphine blocked the increase in dopamine metabolites induced by morphine. These results may be due to the attenuating effect of nalbuphine on the dopaminergic activity of mesolimbic pathways. All of these results suggest nalbuphine could have a great potential as a pharmacotherapy for opiate abuse.