Acquisition Of Morphine-induced Conditioned Place Preference Research Articles

α-Conotoxin TxIB Inhibits Development of Morphine-Induced Conditioned Place Preference in Mice via Blocking α6β2* Nicotinic Acetylcholine Receptors.

Morphine, the main component of opium, is a commonly used analgesic in clinical practice, but its abuse potential limits its clinical application. Nicotinic acetylcholine receptors (nAChRs) in the mesolimbic circuitry play an important role in the rewarding effects of abused drugs. Previous studies have showed that α6β2* (* designated other subunits) nAChRs are mainly distributed in dopaminergic neurons in the midbrain area, which regulates the release of dopamine. So α6β2* nAChRs are regarded as a new target to treat drug abuse. α-Conotoxin TxIB was discovered in our lab, which is the most selective ligand to inhibit α6β2* nAChRs only. Antagonists of α6β2* nAChRs decreased nicotine, cocaine, and ethanol rewarding effects previously. However, their role in morphine addiction has not been reported so far. Thus, it is worth evaluating the effect of α-conotoxin TxIB on the morphine-induced conditioned place preference (CPP) and its behavioral changes in mice. Our results showed that TxIB inhibited expression and acquisition of morphine-induced CPP and did not produce a rewarding effect by itself. Moreover, repeated injections of TxIB have no effect on learning, memory, locomotor activity, and anxiety-like behavior. Therefore, blocking α6/α3β2β3 nAChRs inhibits the development of morphine-induced CPP. α-Conotoxin TxIB may be a potentially useful compound to mitigate the acquisition and/or retention of drug-context associations.

Differential alteration in gut microbiome profiles during acquisition, extinction and reinstatement of morphine-induced CPP

Substance addiction is a chronic and complicated disease involving genetic and environmental factors. Coregulated by the above factors, perturbations of the gut microbiome have been shown to have an essential role in the development of many neuropsychiatric disorders, including addiction. However, shifts in the gut microbiome during different stages of morphine addiction remain uncharacterized. In the present study, we harvested fecal samples from mice at the acquisition (both the control and morphine groups), extinction and reinstatement stages of morphine-induced conditioned place preference (CPP). Gut microbiome profiles were detected with 16S ribosomal RNA gene sequencing. We observed an increase in community richness following morphine conditioning, and it decreased after 4 weeks of abstinence. The abundance of Verrucomicrobia increased and Bacteroides decreased at the acquisition of morphine-induced CPP, while a recovery trend was found at the extinction stage. Several discriminative genera were identified for the characterization of different stages of morphine CPP. Functional analysis of taxa with differential abundance between CPP stages was mainly enriched in the pathways of amino acid metabolism. Taken together, our findings will extend the association between dysbiosis of the gut microbiome and the opioid-induced rewarding or reinforcing behaviors.

Corydaline and l-tetrahydropalmatine attenuate morphine-induced conditioned place preference and the changes in dopamine D2 and GluA1 AMPA receptor expression in rats

Corydalis is a Chinese herb that has been used in China for hundreds of years for analgesic and other purposes. Corydaline and l-tetrahydropalmatine (l-THP) are the main active ingredients of Corydalis. This study was aimed to study the potential utility of corydaline and l-THP in the treatment of opioid abuse and addiction and explore the possible mechanisms underlying their pharmacological actions. Conditioned place preference (CPP) was used to evaluate the rewarding effects of morphine and Western-blot immunoreactive assays were used to evaluate morphine-induced changes in dopamine D2 receptor and GluA1 AMPA receptor and GluA2 AMPA receptor expression in the brain of rats. Systemic administration of corydaline (5 mg/kg, i.p.) or l-THP (1.25, 2.5,5 mg/kg) significantly inhibited the acquisition and expression of morphine-induced CPP in a dose-dependent manner. Corydaline or l-THP alone, at the same doses, failed to produce CPP or conditioned place aversion, and didn't affect locomotor activity. We then examined the expression of dopamine D2 receptor and GluA1 AMPA receptor and GluA2 AMPA receptor subunit expression in rats after acquisition of morphine-induced CPP. We found that repeated administration of morphine produced a significant reduction in dopamine D2 receptor expression in the prefrontal cortex, hippocamps, and striatum, while an increase in the striatal GluA1 AMPA receptor expression. Pretreatment with corydaline or l-THP blocked morphine-induced dopamine D2 receptor down-regulation and GluA1 AMPA receptor up-regulation in these brain regions. Corydaline and l-THP may have therapeutic potential in prevention and treatment of opioid abuse and addiction. The underlying mechanisms may be related to their antagonism on morphine-induced changes in dopamine and glutamate transmission in the brain.

The influence of a new derivate of kisspeptin-10 – Kissorphin (KSO) on the rewarding effects of morphine in the conditioned place preference (CPP) test in male rats

Kissorphin (KSO) is a new peptide derived from kisspeptin-10. Previous study has indicated that this peptide displays neuropeptide FF (NPFF)-like anti-opioid activity. Herein, we examined the influence of KSO (1; 3, and 10 nmol, intravenously [i.v.]), on the rewarding action of morphine (5 mg/kg, intraperitoneally [i.p.]), using the unbiased design of the conditioned place preference (CPP) paradigm in rats. To test the effect of KSO on the acquisition of morphine-induced CPP, KSO and morphine were co-injected during conditioning with no drugs treatment on the test day. To investigate the effect of KSO on the expression of morphine-induced CPP, morphine alone was given during the conditioning phase (1 × 3 days) and KSO was administered 5 min prior to the placement in the CPP apparatus on the test day. To estimate the influence of KSO on the reinstatement of morphine-induced CPP, KSO was given 5 min before a priming dose of morphine (5 mg/kg, i.p.) on the reinstatement test day. The results show that KSO inhibited the acquisition, expression and reinstatement of morphine-induced CPP. The strongest effect of KSO was observed at the dose of 10 nmol (acquisition and reinstatement) or 1 nmol (expression). KSO given alone, neither induced place preference, nor aversion. Furthermore, the morphine-modulating effects of KSO were markedly antagonized by pretreatment with RF9 (10 nmol, i.v.), the NPFF receptors selective antagonist. Thus, KSO inhibited the morphine-induced CPP mainly by involving specific activation of NPFF receptors. Overall, these data further support the anti-opioid character of KSO.

An Assessment between D1 Receptor Agonist and D2 receptor Antagonist into the Ventral Tegmental Area on Conditioned Place Preference and Locomotor Activity.

Background:The release of dopamine (DA) has certain roles in the induction of conditioned place preference (CPP) and motor learning in the ventral tegmental area (VTA). The aim of this study was to investigate the excitatory effects of DA through DA-D1 agonist (SKF38393) and elimination of the inhibitory effects of DA through DA-D2 antagonist (eticlopride) into the VTA and its synergistic effects with an ineffective dose of morphine in the induction of CPP.Materials and Methods:Morphine (2.5 mg/kg; s. c.) did not induce a significant CPP, without any effect on the locomotor activity during the testing phase. SKF38393 (0.125, 0.5, and 1 μg/side) and eticlopride (0.5, 1, and 2 μg/side) individually or simultaneously were microinjected bilaterally into the VTA.Results:The administration of SKF38393 (1 and 2 μg/rat) with ineffective morphine and also without morphine caused CPP on test day, while eticlopride (2 μg/rat) caused CPP with morphine only. Locomotor activity increased in groups receiving D1 agonist and D2 antagonist that presumed to be caused by the reinforcing effect. In addition, the concurrent administration of ineffective doses of D1 agonist and D2 antagonist into the VTA with ineffective morphine caused CPP but not with saline.Conclusions:This study showed that there was a need for morphine to activate the reward circuit through the D2 receptor in the VTA while the administration of the D1 agonist could independently activate the reward circuit. In addition, there was a probable synergistic effect using ineffective doses of D1 and D2 receptors, in the acquisition of morphine-induced CPP.

Inhibitory effects of forced swim stress and corticosterone on the acquisition but not expression of morphine-induced conditioned place preference: Involvement of glucocorticoid receptor in the basolateral amygdala

Addiction is a common chronic psychiatric disease which represents a global problem and stress has an important role to increase drug addiction and relapse. In the present study, we investigated the effects of physical stress and exogenous corticosterone on the acquisition and expression of morphine-induced conditioned place preference (CPP). Also, we tried to find out the role of glucocorticoid receptors (GRs) of basolateral amygdala (BLA) in this regard. In the CPP paradigm, conditioning score and locomotion activity were recorded by Ethovision software. Male adult rats received forced swim stress (FSS) as a physical stress or corticosterone (10mg/kg; ip) as a dominant stress hormone in rodents, 10min before morphine injection (5mg/kg; sc) during three conditioning days (acquisition) or just prior to CPP test in the post-conditioning day (expression). In FSS procedure, animals were forced to swim for 6min in cylinder filled with water (24–27°C). To evaluate the role of glucocorticoid receptors in the BLA, different doses of mifepristone (RU38486) as a GR antagonist were injected into the BLA (0.3, 3 and 30ng/side) during 3-day conditioning phase before FSS or injection of corticosterone in morphine-CPP paradigm. The results showed that FSS and corticosterone reduce the acquisition but not expression of morphine-induced CPP. Moreover, blockade of GRs in the BLA could diminish the inhibitory effects of FSS or corticosterone on the acquisition of morphine-induced CPP. It seems that stress exerts its effect on reward pathway via glucocorticoid receptors in the BLA.

Streptozotocin-induced diabetes affects the development and maintenance of morphine reward in rats

Diabetes mellitus is a chronic illness that has been associated with the decrease of insulin in type I diabetes. Insulin has an impact not only on the direct control of food intake and plasma glucose levels, but also on brain pathways associated with reward. It affects brain reward pathways through regulation of the dopamine (DA) transporter (DAT). Moreover, it has been found to affect the ability of drugs that target the DA system. In the present study, the effects of streptozotocin (STZ)-induced diabetes on the acquisition (development) and maintenance of morphine-induced conditioned place preference (CPP) were investigated in rats. Forty adult male Albino Wistar rats were used in these experiments. For induction of diabetes, STZ was administered at a dose of 60mg/kg. After seven days, the CPP paradigm was done; conditioning score and locomotor activity were recorded by Ethovision software. The results showed that diabetes significantly increased the magnitude of conditioning scores, acquisition of morphine-induced CPP in compared to naive animals (P<0.05). Moreover, in the diabetic group, there were significant differences among conditioning scores in the post-conditioning phase and the last four days (7th–10th), but these differences elongated up to 10 days after the CPP protocol while the extinction period was eight days in the naive group. Our findings indicated that the magnitude and maintenance of morphine rewarding properties have been changed in STZ-induced diabetic animals. It seems that a level of insulin and their receptors are involved in the development and maintenance of morphine-induced CPP in the rats.

Involvement of CB1 receptors in the ventral tegmental area in the potentiation of morphine rewarding properties in acquisition but not expression in the conditioned place preference model

The ventral tegmental area (VTA) is a critical part of the brain reward system and has been engaged in mediating rewarding actions. CB1 receptors are one of the receptors that mediate the actions of cannabinoids and endocannabinoids in the central nervous system. Our aim was to determine the potentiating effects of CB1 receptors within the VTA in the acquisition and expression of morphine conditioned place preference (CPP). Stereotaxic surgery was performed bilaterally on each rat to administrate WIN55,212-2 (1, 2 and 4mmol/0.3μl DMSO) as CB1 receptor agonist and AM251 (15, 45 and 90mmol/0.3μl DMSO) as CB1 receptor antagonist. A three-compartment apparatus was used for the CPP test. The results showed that two doses of WIN55,212-2 (2 and 4mmol) potentiates the rewarding effects of ineffective dose of morphine (2mg/kg). We did not see any significant difference between any other doses of WIN55,212-2 and vehicle in the group which received the effective dose of morphine (5mg/kg). Additionally, conditioning scores decreased significantly with the highest administrated dose of AM251 (90mmol) compared to the vehicle group. We did not observe any significant differences in the experiments for CPP expression by WIN55,212-2 or AM251. It seems that the cannabinoid and opioid systems are in interaction with each other and affect dopaminergic and/or non-dopaminergic neurons in the VTA. Blockade of CB1 receptors may increase GABA release, resulting in the reduction of dopamine output followed by a decrease in the acquisition of morphine-induced CPP in rats.

Involvement of p38/NF-κB signaling pathway in the nucleus accumbens in the rewarding effects of morphine in rats

The nucleus accumbens (NAc) is one of the important brain regions for the acquisition of morphine-induced conditioned place preference (CPP), which is a valuable paradigm for detecting the rewarding effects of drugs. However, the underlying molecular mechanisms remain largely unknown. In the present study, we investigated the role of p38 and nuclear factor kappa B (NF-κB) in the NAc in the acquisition of morphine-induced CPP. The results showed that repeated morphine treatment induced the acquisition of CPP and increased the phosphorylation of p38, IκB-α and NF-κB p65 in the NAc. Microinjection of p38 inhibitor SB203580 into the NAc prior to the application of morphine prevented the acquisition of CPP and inhibited the activation of p38, IκB-α and NF-κB p65. Furthermore, pre-infusion of pyrrolidine dithiocarbamate (PDTC), a selective NF-κB inhibitor, into the NAc blocked the CPP but also the phosphorylation of NF-κB p65 induced by morphine. It is concluded that the activation of p38/NF-κB p65 signaling pathway in the NAc plays a critical role in the morphine CPP.

Activation of phosphatidylinositol 3-kinase/Akt-mammalian target of Rapamycin signaling pathway in the hippocampus is essential for the acquisition of morphine-induced place preference in rats

Hippocampus is a critical structure for the acquisition of morphine-induced conditioned place preference (CPP), which is a usual learning paradigm for assessing drug reward. However, the precise mechanisms remain largely unknown. Phosphatidylinositol 3-kinase (PI3K) and its downstream targets, including Akt, mammalian target of Rapamycin (mTOR) and 70-kDa ribosomal S6 kinase (p70S6K), are critical molecules implicated in learning and memory. Here, we tested the role of PI3K/Akt-mTOR-p70S6K signaling pathway in morphine-induced CPP in the hippocampus. Our results showed that the acquisition of morphine CPP increased phosphorylation of Akt in the hippocampal CA3, but not in the nucleus accumbens (NAc), the ventral tegmental area (VTA) or the CA1. Moreover, the phosphorylated Akt exclusively expressed in the CA3 neurons. Likewise, levels of phosphorylated mTOR and p70S6K were significantly enhanced in the CA3 following morphine CPP. The alterations of these phosphorylated proteins are positively correlated with the acquisition of morphine CPP. More importantly, microinjection of PI3K inhibitor (LY294002) or mTOR inhibitor (Rapamycin) into the CA3 prevented the acquisition of CPP and inhibited the activation of PI3K-Akt signaling pathway. In addition, pre-infusion of β-FNA (β-funaltrexamine hydrochloride), a selective irreversible μ opioid receptor antagonist, into CA3 significantly prevented the acquisition of CPP and impaired Akt phosphorylation. All these results strongly implied that the PI3K-Akt signaling pathway activated by μ opioid receptor in hippocampal CA3 plays an important role in acquisition of morphine-induced CPP.

Effects of reversible inactivation of the ventral tegmental area on the acquisition and expression of morphine-induced conditioned place preference in the rat

The mesolimbic dopaminergic system that projects from the ventral tegmental area (VTA) to the nucleus accumbens is critical for initiation of opioid reinforcement and reward-related effects of drugs of abuse. In the present study, the effects of reversible inactivation of VTA on acquisition and expression of morphine-induced conditioned place preference (CPP) were investigated in rats. One hundred and eighty-one adult male albino Wistar rats were used in these experiments. Reversible inactivation of VTA was done through the unilateral and bilateral intra-VTA microinjection of 2% lidocaine during the acquisition and expression of morphine (5 mg/kg; s.c.)-induced CPP. Animal displacement, conditioning score and locomotor activity were recorded by Ethovision software. The results showed that bilateral but not unilateral intra-VTA administration of lidocaine significantly decreases the acquisition ( P < 0.01) and expression ( P < 0.05) of morphine-induced CPP compared to their respective saline-microinjected groups. Moreover, intra-VTA administration of lidocaine had no effect on locomotor activity in these experiments. Our results further support the idea that VTA may play an important role in the acquisition of morphine-induced CPP. In addition, there is no functional lateralization in the VTA for acquisition and/or expression of morphine-induced CPP in the rat.

Blockade of cue- and drug-induced reinstatement of morphine-induced conditioned place preference with intermittent sucrose intake

Sucrose intake has been suggested to alter the expression of morphine-induced conditioned place preference (CPP). To date, the potential effects of sucrose intake on the extinction and drug-induced reinstatement of CPP have not been determined. In the present study, sucrose solution (15%) was given prior to, during, and following the acquisition of morphine-induced CPP. Place preference was subsequently assessed during expression, extinction, and morphine-induced reinstatement. The results showed that the sucrose solution given prior to place conditioning training transiently suppressed the expression of morphine CPP. Sucrose solution given during place conditioning training had no effects on the expression, extinction, and reinstatement of CPP. When the sucrose solution was given following the acquisition of morphine CPP, the extinction of morphine CPP was accelerated, and morphine-induced reinstatement was profoundly inhibited. The above results demonstrated that sucrose intake could differentially affect the expression, extinction, and reinstatement of morphine-induced CPP, depending on the interference schedules. Our findings suggest that offering non-drug rewards could be a valuable approach to maintain abstinence and preventing relapse in drug addicts.

Dopamine D3 receptor ligands modulate the acquisition of morphine-conditioned place preference.

The dopamine D3 receptor has been shown to mediate conditioned effects of psychostimulants such as cocaine. The present work was aimed at determining whether drugs acting at D3 receptors alter acquisition of conditioned effects of opiates. We have used the conditioned place preference (CPP) in mice, which permits the measurement of approach behaviour to environmental stimuli previously paired with drug effects. To assess the interaction of morphine and D3 receptor ligands during acquisition of CPP, we have used a particular procedure, in which the animals were given the choice between compartments associated with either morphine alone or the combination of morphine with the tested agent. D3 receptor agonists (7-OH-DPAT, quinelorane, BP 897) did not induce, alone, a significant CPP but, all of them, at the doses tested, and notably BP 897, a highly selective partial agonist, significantly enhanced acquisition of morphine-induced CPP when administered together with morphine at each conditioning session. PNU-99194A, a D3 receptor-preferring antagonist, induced a CPP itself at the dose of 10 mg/kg but not at 5 or 15 mg/kg and impaired significantly at 10 and 15 mg/kg the morphine-induced CPP. In contrast, BP 897 did not alter morphine-induced analgesia, an unconditioned effect of this drug. These results suggest the stimulation of D3 receptors has no rewarding effect per se, but may synergize upon opiate-induced dopamine release with stimulation of other dopamine receptor subtypes to enhance approach behaviour to morphine-associated environment.