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Official websites use. Share sensitive information only on official, secure websites. To whom correspondence should be addressed. E-mail: ocivelli uci. Edited by Solomon H. Author contributions: S. Drug addiction is mediated by complex neuronal processes that converge on the shell of the nucleus accumbens NAcSh. Also, injecting MCH potentiates cocaine-induced hyperactivity in mice. Mice lacking MCH1R exhibit decreased cocaine-induced conditioned place preference, as well as cocaine sensitization. Using a specific MCH1R antagonist, we further show that acute blockade of the MCH system not only reduces cocaine self-administration, but also attenuates cue- and cocaine-induced reinstatement. Thus, the MCH system has an important modulatory role in cocaine reward and reinforcement by potentiating the dopaminergic system in the NAcSh, which may provide a new rationale for treating cocaine addiction. The shell of the nucleus accumbens NAcSh is known to be an important center for reward and motivation. It is a major terminal area of the mesolimbic dopamingergic system, which projects from the ventral tegmental area VTA 1. The NAcSh receives inputs from different brain areas, in particular, from the lateral hypothalamus LH 3. The importance of the LH in reward and motivation has been shown by Olds 4 , who discovered that electrical activation of the LH induced an extraordinarily intense self-stimulation response in rats. However, the transmitter s responsible for this action have not been established definitively. MCH is a cyclic, amino acid peptide originally isolated from salmon pituitary as a melanophore-concentrating factor It is also present in rats and humans 11 , 12 , where it is viewed as important in regulating nutritional homeostasis 8 , Indeed, the effects of the MCH system on feeding and mood behaviors already have been shown to depend on its activity in the NAcSh This report provides evidence for this hypothesis by showing that the dopamine and MCH systems can interact in the NAcSh at the level of their receptors, and that MCH administration can increase dopamine responses in vitro and in vivo. Also, chronic blockade of the MCH system attenuates motivation for cocaine, whereas acute blockade inhibits cocaine self-administration, as well as cue- and cocaine-induced reinstatement. Therefore, our results suggest that the MCH system is a powerful potentiator of the dopaminergic system in NAcSh neurons, exhibiting an important modulatory role in cocaine reward and reinforcement, which may provide a new rationale for treating cocaine addiction. Previously, we and others showed that MCH1R is expressed in many areas of the brain 9 , Our localization data suggest that the MCH system in the NAcSh may modulate dopamine-related responses and dopamine-mediated addictive behaviors. To understand whether the MCH system interacts functionally with the dopamine system in the NAcSh, we used whole-cell patch-clamp electrophysiology to monitor spike firing in NAcSh neurons. Application of MCH 2. These results indicate that the MCH and dopamine systems can interact to elevate spike firing in the NAcSh, and that blockade of the MCH system would inhibit that increase in spike firing. This hypothesis was confirmed by using a biochemical approach. Together, these experiments led us to conclude that MCH and dopamine receptors can act in concert to enhance the activity of NAcSh neurons. To analyze whether MCH system activation can potentiate the dopamine system in vivo, we investigated whether cocaine-induced hyperactivity is enhanced by exogenous MCH administration. Mice were injected either with vehicle or MCH 0. Time course of horizontal activity after vehicle, MCH 1 nmol i. However, i. Effect of MCH on cocaine induced locomotor activity. B Time course of horizontal activity after vehicle, MCH 1 nmol i. C Cumulative horizontal activity. To test whether the endogenous MCH system contributes to dopamine-mediated reward in vivo, MCH1R KO mice were tested for their cocaine-seeking behavior in the conditioned place preference CPP paradigm and for any changes in their locomotor activity during cocaine sensitization. We used a 2-chamber, nonbiased, CPP model. Mice were injected either with vehicle or cocaine on alternate days, and confined in one of the chambers during conditioning sessions. During the postconditioning session, increased time spent in the cocaine-paired chamber was taken to indicate increased cocaine-seeking behavior. We found that cocaine induced CPP in WT mice, as shown by the increase in the time spent in the cocaine-associated compartment during the postconditioning session Fig. The data were normalized to saline treated WT group a. MCH1R KO mice also were tested for sensitization of locomotor activity in response to repeated cocaine administration. WT mice showed significantly increased locomotor activity after repeated daily cocaine injections Fig. Together, these results indicate that the lack of MCH1R results in a decreased sensitivity to the rewarding properties of cocaine in vivo. Therefore, MCH system activation is involved in the rewarding effect of cocaine. Chronic exposure to cocaine has been shown to increase DARPP phosphorylation at Thr 75 in the caudate putamen and nucleus accumbens In contrast, we found that MCH1R KO mice do not exhibit a similar change in phosphorylation level after chronic cocaine treatment. Cocaine self-administration is the most common paradigm for studying drugs of abuse. We studied how pharmacological blockade of the MCH system affects cocaine self-administration. This antagonist inhibits MCH1R with a nanomolar affinity, it is highly specific, showing no affinity for a panel of receptors and channels, is active in vivo, and does not exert unwanted side effects The cocaine self-administration paradigm measures the primary reinforcing effect of cocaine. Animals were trained to self-administer cocaine 0. Vehicle or TPI 5, 10, or 20 nmol, i. TPI also was tested in rats working on a progressive ratio PR schedule. Under this schedule, rats are required to increase the number of nose-pokes required for each successive cocaine injection. The breakpoint was defined as the number of completed nose pokes before a min period when no infusions were obtained by the rat. An increase in the breakpoint after TPI injection indicates that blockade of the MCH system increases the reinforcing properties of cocaine, whereas a decrease indicates that MCH blockade reduces the motivation for cocaine. The steps of the exponential progression were the same as those used previously As shown in Fig. The decrease in PR breakpoint suggests that blockade of the MCH system leads to a reduction in the rewarding efficacy of cocaine and in motivational properties for cocaine-taking behavior. Effect of MCH system blockade on cocaine self-administration. A TPI 5—20 nmol i. To analyze this possibility, we first tested whether TPI elicits its inhibitory action by acting in the NAcSh. Relapse is an important aspect of drug addiction It is generally studied after cocaine self-administration and then extinction of response to cocaine; after this time, cocaine response can be reinstated by exposing the animals to cocaine-associated cues, priming injection of cocaine or stress. In our experiments, the cue was a house light that predicted drug availability and a 5. In these tests, the house light was turned on at the start of the session, and every nose poke resulted in activation of the discrete cue light, but not in cocaine delivery. Effect of MCH system blockade on cue-, cocaine-, and stress-induced reinstatements. C Stress-induced reinstatement: TPI inhibition of reinstatement induced by priming injection of yohimbine 2. Next, we determined the effect of TPI on reinstatement of cocaine-seeking primed by injections of cocaine. Cocaine self-administration training and extinction were carried out as described above. These 2 experiments indicate that the endogenous MCH system is critical for relapse induced by cocaine-associated cues, as well as a priming injection of cocaine. Another stimulus that triggers relapse-like behavior in rats is brief exposure to stress. Therefore, we tested the effect of TPI on stress-induced reinstatement of cocaine seeking. Priming injections of yohimbine 2. Thus, MCH1R activation does not appear to contribute to relapse induced by a stressor. Together, our results strongly suggest that endogenous MCH has a critical role during reinstatement of cocaine-seeking by a cocaine-associated cue or by a cocaine priming injection, but not by stress. The MCH system has been implicated in modulating several behavioral responses 13 ; most notably, MCH has been shown to induce increases in food intake when administered centrally 8. In this article, we report another role for the MCH system related to its receptor expression in the NAcSh: namely, a modulatory role on dopamine-related responses and, in particular, on cocaine addiction. This data is in agreement with the data of Georgescu et al. Although the high degree of overlap found in both studies points toward coexpression of D1R and D2R in medium spiny neurons, it should be mentioned that this is still a matter of controversy 28 , and that using MCH1R, D1R, and D2R triple in situ hybridization analyses could resolve this issue. In vivo data have shown that several motivated behaviors require both D1 and D2 receptor activation in the NAc For example, dopamine-induced spike firing in NAcSh neurons requires activation of both D1 and D2 receptors We show here that MCH can modulate this response, and that its effects support the need for D1 and D2 coactivation. MCH had no significant effect on spike firing in NAcSh neurons, whether alone or together with one of the D1 or D2 agonists, although it had been previously shown to decrease D1R induced GluR1 phosphorylation This difference may be due to differences in the methodologies GluR1 phosphorylation vs. However, from our data, only when D1 and D2 are coactivated is observed the potentiating effect of MCH in spike firing. The modulatory role of MCH on the dopamine system was further investigated at the cellular level. DARPP is known to be a central target of dopamine-induced intracellular responses This potentiating effect of MCH is consistent with the one discovered through our electrophysiological observations. Our data support the view that DARPP phosphorylation at Thr 75 has a role in mediating the development of cocaine sensitization, as already shown by the fact that Thr75Ala DARPP mutant mice do not exhibit cocaine sensitization Therefore, these results show that MCH system activation has a potentiating effect on the responses that dopamine induces in the NAcSh. Next, we explored whether MCH system activation can potentiate dopamine effects in vivo. We tested whether MCH injections can enhance cocaine-induced hyperactivity. Thus, injection of exogenous MCH can potentiate a cocaine effect known to be induced by increased dopamine concentration in the nerve terminals. These in vivo data are consistent with our in vitro data, and show that the MCH system can potentiate dopamine effects. These mice exhibited attenuated cocaine-induced CPP and a blunted development of cocaine sensitization, which suggests that MCH system inhibition decreases the rewarding effects of cocaine. Tyhon et al. This discrepancy may be explained by differences in procedures. CPP under this biased design requires the animal to also overcome the aversion or anxiety toward the nonpreferred compartment 34 — Therefore, the CPP score under a biased CPP procedure represents not only the motivation for cocaine, but also the overcoming of the anxiety experienced in the conditioning compartment. However, the unbiased design used in our CPP experiments excludes the anxiety factor, an important factor, because the MCH system has been reported to regulate anxiety 37 , 38 , and MCH1RKO mice tend to be less anxious Therefore, we suggest that the differences in CPP results derive from differences in the designs of the assays. Our data also could be seen to conflict with studies that have reported that MCH1R KO mice are hypersensitive to amphetamine 39 , 40 , which is thought to act via the same mechanism as cocaine. However, some evidence suggests that cocaine and amphetamine induce hyperactivity via different mechanisms. The effect of cocaine on dopamine accumulation depends on the firing of dopaminergic neurons, whereas the effects of amphetamine do not 41 , Other KO mice strains, such as the NMDA receptor-deficient mice NR1-KD , did not exhibit hyperactivity on acute cocaine administration 43 while their amphetamine response was not altered. Therefore, it is possible that the effects of cocaine on behavior are distinct from those of amphetamines. KO mice represent a chronically inhibited model that does not allow one to study the effects that acute blockade of a neuronal system has on drug self-administration and drug relapse. Using an MCH1R antagonist, we show that acute blockade of the MCH system reduces cocaine self-administration in a dose-dependent manner. This effect is seen when the antagonist is administered in the shell, but not in the core of the NAc. However, this effect may be due to either an increase in cocaine reinforcing effect or a decrease in motivation for cocaine 25 , To differentiate between these 2 possibilities, we tested whether acute blockade of the MCH system can affect cocaine self-administration under a PR schedule. Although this decrease is smaller than that found for other compounds that are able to inhibit cocaine reward 45 — 47 , it is significant, which indicates that blockade of the MCH system does not increase the reinforcing effect of cocaine. Therefore, TPI decreases motivation to self-administer cocaine. These data strongly support the results we obtained with mice lacking MCH1R. The inhibitory effect of TPI on cocaine self-administration, a dopamine-induced response, may seem contrary to the increased performance that another MCH1R antagonist, peptide 30 has in the forced swim test 19 , a model where reduced dopamine levels correlate with increased depression-like state. However, it should be noted that the physiology of depression, more than that of cocaine addiction, involves the activities of neurotransmitter systems others than dopamine 48 , and that, for unknown reasons, different MCH1R antagonists exert variable performance in antidepressant-like assays There is considerable interest in the cellular mechanisms of reinstatement of drug-seeking. Our data add another player in these mechanisms by showing that acute blockade of the MCH system significantly reduces relapse to cocaine-seeking that is induced by cocaine or by a conditioned cue, but not by a stressor. Previous studies have shown that the relapse induced by a conditioned cue or by cocaine activates the mesolimbic dopamine system, whereas the relapse induced by stress involves activities of the corticotrophin-releasing factor CRF and of the noradrenergic NE system 50 , Therefore, our data indicate that the mesolimbic dopamine system is most directly related to the role of the MCH system in reinstatement. Together, our data allow us to conclude that blockade of the endogenous MCH system inhibits cocaine taking and seeking, in agreement with our discovery that activation of the MCH system potentiates the dopamine system to induce cellular responses in NAcSh. Thus, a specific MCH1R antagonist may represent a previously undescribed therapeutic tool for treating cocaine addiction or other disorders that result from dopamine imbalances in the NAcSh. Detailed methods are described in the SI Materials and Methods. Male Sprague—Dawley Charles River rats were used for all of the other experiments. Electrophysiology experiments were performed by using whole-cell recording as described previously CPP was conducted in 2-chambered boxes containing visual, olfactory, and tactile cues with the unbiased method for drug conditioning see SI Materials and Methods. The time spent in each compartment during pre and postconditioning tests 20 min was measured. Response in the active hole resulted in illumination of the signal light 5. Response in the inactive hole was counted, but had no consequences see SI Materials and Methods. We thank Dr. Paul Greengard, Dr. Su Qian of Merck. This article contains supporting information online at www. This section collects any data citations, data availability statements, or supplementary materials included in this article. As a library, NLM provides access to scientific literature. The melanin-concentrating hormone system modulates cocaine reward Shinjae Chung Shinjae Chung Departments of a Pharmacology and. Find articles by Shinjae Chung. Find articles by F Woodward Hopf. Find articles by Hiroshi Nagasaki. Chun-Ying Li Departments of a Pharmacology and. Find articles by Chun-Ying Li. James D Belluzzi Departments of a Pharmacology and. Find articles by James D Belluzzi. Find articles by Antonello Bonci. Olivier Civelli Departments of a Pharmacology and. Find articles by Olivier Civelli. Departments of a Pharmacology and. Received Nov 10; Issue date Apr PMC Copyright notice. Open in a new tab. The authors declare no conflict of interest. Similar articles. Add to Collections. Create a new collection. Add to an existing collection. Choose a collection Unable to load your collection due to an error Please try again. Add Cancel.