Dopaminergic and nitric oxide systems interact to regulate the electrical activity of neurons in the medial septal nucleus in rats.

Research characterizing the neuronal substrate of anxiety has implicated different brain areas, including the medial septal nucleus (m-SEPT). Previous reports indicated a role of dopamine and nitric oxide (NO) in anxiety-related behaviors. In this study, the extracellular single-unit recording was performed from the m-SEPT in adult male albino Wistar rats. Baseline activity was recorded for 5 min, and the post-injection recording was performed for another 5 min after the microinjection of each drug. The results showed that (1) both D1- and D2-like receptor agonists (SKF-38393 and quinpirole) enhanced the firing rate of m-SEPT neurons; (2) both D1- and D2-like antagonists (SCH-23390 and sulpiride) attenuated the firing rate of m-SEPT neurons; (3) L-arginine (NO precursor) increased the firing rate of m-SEPT neurons, but a non-specific NOS inhibitor, L-NAME, elicited no significant alterations; (4) the non-specific NOS inhibitor reversed the enhanced firing rate produced by SKF-38393 and quinpirole; (5) neither of the dopaminergic antagonists changed the enhanced activity resulted from the application of the NO precursor. These results contribute to our understanding of the complex neurotransmitter interactions in the m-SEPT and showed that both dopaminergic and NO neurotransmission are involved in the modulation of the firing rate of neurons in the m-SEPT.

Differential Roles of Intra-accumbal Orexin Receptors in Acquisition and Expression of Methamphetamine-Induced Conditioned Place Preference in the Rats.

A large amount of document has revealed that the orexin system in the reward circuity, including the nucleus accumbens (NAc), contributes to the modification of drug reinforcement. It has proven that the orexin receptors (OXRs) are expressed on dopamine terminals in the NAc; therefore, it can modulate reward-related behaviors. In the present study, the conditioned place preference (CPP) paradigm was used to evaluate the role of OXRs in the NAc in the acquisition and expression of methamphetamine (METH)-induced CPP. Based on previous studies, animals received METH (1 mg/kg; sc) on a 5-day schedule to induce CPP. The rats bilaterally received SB334867, OX1R antagonist, or TCS OX2 29, OX2R antagonist, (1, 10, and 30 nM/0.5 µl DMSO 12%) over five days of conditioning by METH to display the role of OXRs in reward acquisition. Moreover, the rats bilaterally received SB334867 or TCS OX2 29 in the NAc before the post-conditioning test to consider the impact of OXR antagonists on the expression of METH-induced CPP. The data revealed that the administration of SB334867 or TCS OX2 29 in the NAc led to a decrease in the acquisition of METH-induced CPP. Additionally, intra-accumbal injection of OX1R antagonist inhibited the expression of METH-induced CPP, while the OX2R antagonist failed to change this expression. Finally, the intra-NAc microinjection of both OXR antagonists was more effective in inhibiting acquisition than blocking the expression phase of METH. Data from the current study confirms that OXRs in the NAc regulate the reward-related effects of METH.

D1- but not D2-like dopamine receptor antagonist in the CA1 region of the hippocampus reduced stress-induced reinstatement in extinguished morphine-conditioning place preference in the food-deprived rats.

Relapsing to drugs of abuse is a challenging problem in treatment of addiction and stress is believed to be a major risk factor in relapse to drugs. The hippocampus region and dopamine signaling play a critical role in reward-related behaviors. The purpose of this study is to identify the involvement of D1- and D2-like receptors in the CA1 region of hippocampus in the reinstatement induced by a combination of food deprivation stress and a sub-threshold dose of morphine in extinguished morphine-conditioning place preference in rats. Adult male rats treated with one specific doses of SCH-23390 or sulpiride (0.5, 2 and 4 µg/0.5 µl vehicle/side) as D1- and D2-like receptors antagonists into the CA1 in separate groups, following the conditioning and extinction phase of morphine-conditioning place preference, before initiating the food deprivation stress on the last day of extinction. Then, the food deprived animals examined for reinstatement by injection of the sub-threshold dose of morphine (0.5 mg/kg, s.c.) on reinstatement day. Conditioning place preference scores and locomotor activities were recorded during test. Our results showed that combination of food deprivation stress and a sub-threshold dose of morphine induced the reinstatement of morphine-conditioning place preference. The induced reinstatement was decreased by two higher doses of SCH-23390 (2 and 4 µg/0.5 µl vehicle/side). However, the sulpiride (0.5, 2 and 4 µg/0.5 µl vehicle/side) could not reduce the reinstatement. Results showed that the role of D1-like receptor in the CA1 region was more prominent than D2-like receptor in reinstatement induced by food deprivation stress and re-exposure to morphine. Therefore the D1-like receptor in the CA1 might be a potential therapeutic target for treatment of opiate addiction.

Selective Modulation of Hippocampal Theta Oscillations in Response to Morphine versus Natural Reward.

Despite the overlapping neural circuits underlying natural and drug rewards, several studies have suggested different behavioral and neurochemical mechanisms in response to drug vs. natural rewards. The strong link between hippocampal theta oscillations (4-12 Hz) and reward-associated learning and memory has raised the hypothesis that this rhythm in hippocampal CA1 might be differently modulated by drug- and natural-conditioned place preference (CPP). Time-frequency analysis of recorded local field potentials (LFPs) from the CA1 of freely moving male rats previously exposed to a natural (in this case, food), drug (in this case, morphine), or saline (control) reward cue in the CPP paradigm showed that the hippocampal CA1 theta activity represents a different pattern for entrance to the rewarded compared to unrewarded compartment during the post-test session of morphine- and natural-CPP. Comparing LFP activity in the CA1 between the saline and morphine/natural groups showed that the maximum theta power occurred before entering the unrewarded compartment and after the entrance to the rewarded compartment in morphine and natural groups, respectively. In conclusion, our findings suggest that drug and natural rewards could differently affect the theta dynamic in the hippocampal CA1 region during reward-associated learning and contextual cueing in the CPP paradigm.

Differential Aspects of Natural and Morphine Reward-related Behaviors in Conditioned Place Preference Paradigm.

Introduction: Natural rewards are essential for survival. However, drug-seeking behaviors can be maladaptive and endanger survival. The present study was conducted to enhance our understanding of how animals respond to food and morphine as natural and drug rewards, respectively, in a conditioned place preference (CPP) paradigm. Methods: We designed a protocol to induce food CPP and compare it as a natural reward with morphine CPP in rats. The protocol for reward induction in both groups (foods and morphine) consisted of three phases: pre-test, conditioning, and post-test. In morphine groups, we injected morphine as a reward (5 mg/kg, SC). To induce natural reward, we used two different protocols. In the first one, the rats were deprived of food for 24 h. In the other method, the rats were restricted to food for 14 days. During the conditioning period, the animals received daily chow, biscuits, or popcorn as a reward inducer. Results: Results revealed that CPP was not induced in food-deprived rats. A combination of food restriction (as a facilitator) and a biscuit or popcorn-induced reward using CPP. In contrast, food deprivation did not facilitate food CPP in response to regular food. Interestingly the CPP score of the group which received biscuits during a 7-day conditioning period was more than that of the morphine group. Conclusion: In conclusion, food restriction could be a better protocol than food deprivation to facilitate food reward.

Acute morphine administration, morphine dependence, and naloxone-induced withdrawal syndrome affect the resting-state functional connectivity and Local Field Potentials of the rat prefrontal cortex.

Opiates are among the widely abused substances worldwide. Also, the clinical use of opioids can cause unwanted and potentially severe consequences such as developing tolerance and dependence. This study simultaneously measured the changes induced after morphine dependence and naloxone-induced withdrawal syndrome on the resting-state functional connectivity (rsFC) and Local Field Potential (LFP) power in the prefrontal cortex of the rat. The obtained results revealed that acute morphine administration significantly increased the LFP power in all frequency bands, as well as the rsFC strength of the prefrontal cortex, and naloxone injection reversed this effect. In contrast, chronic morphine administration reduced neural activity and general correlation values in intrinsic signals, as well as the LFP power in all frequency bands. In morphine-dependent rats, after each morphine administration, the LFP power in all frequency bands and the rsFC strength of the prefrontal cortex were increased, and these effects were further enhanced after naloxone precipitated withdrawal syndrome. The present study concludes that general correlation merely reflects the field activity of the local cortices imaged.

Similar role of mPFC orexin-1 receptors in the acquisition and expression of morphine- and food-induced conditioned place preference in male rats.

Self-control problems are a typical character of drug addiction and excessive food consumption and it has been shown that natural rewards and drugs of abuse share parts of the same neural substrate and reward processing in the brain. Different brain areas are involved in natural and drug reward processing including the mesolimbic pathway, amygdala, nucleus accumbens (NAc), and prefrontal cortex. Considering the important role of orexins in the addictive behavior and the presence of orexin-1 subtype receptors (Orx1R) in the medial prefrontal cortex (mPFC), this study investigated the role of mPFC in natural- and drug-reward seeking behaviors to deepen our understanding of possible similarities or differences. To induce food- or morphine-conditioned place preference (CPP), adult male Wistar rats underwent CPP testing and received intra-mPFC doses of SB334867 (3, 10, or 30 nM/0.5 µl DMSO 12%), as an Orx1R antagonist, during the acquisition or expression phases of the CPP test. Results indicated that microinjection of Orx1R antagonist into the mPFC had similar effects on both morphine- and food-induced CPP and attenuated CPP scores in the acquisition and expression phases of the CPP test. The data demonstrated that Orx1Rs in the mPFC regulate the reward-related effects of morphine- and food-induced reward.

Cannabidiol modulates the METH-induced conditioned place preference through D2-like dopamine receptors in the hippocampal CA1 region.

The main problem with addiction is a relapse with a high rate in methamphetamine (METH) abusers. Using addictive drugs repetitively will cause the reward. METH reward is due to an increase in dopamine levels, and the endocannabinoid system (ECS) has a modulatory role in reward through CB1 receptors. On the other hand, the hippocampus plays an important role in learning and memory, so it is involved in the neuroplasticity caused by METH abuse. Cannabidiol (CBD) has been shown to reduce the effects of METH through different mechanisms such as increasing the ECS activity, regulating emotional memory in the ventral hippocampus through D2-like dopamine receptors, and decreasing the mesolimbic dopaminergic activity. The present study tried to find out the role of hippocampal CA1 D2-like dopamine receptors (D2R) in the effects of cannabidiol on the acquisition and expression of METH-induced conditioned place preference (METH-CPP) in rats by using microinjection of sulpiride as a D2R antagonist. For this purpose, different groups of animals received different doses of sulpiride (0.25, 1, and 4 µg/0.5 µL DMSO; CA1), once prior to the injection of CBD (10 µg/5 µL for acquisition and 50 µg/5 µL for expression; ICV) and once in the absence of CBD. Control groups were also considered. In brief, findings showed that cannabidiol decreases METH-induced CPP. Intra-CA1 administration of sulpiride reversed the decreasing effects of cannabidiol on METH-induced CPP in both acquisition and expression phases but more prominent in the expression phase. The results showed that sulpiride did not affect the METH-induced CPP in the absence of cannabidiol. In conclusion, this study demonstrated that cannabidiol decreased METH-induced CPP in part through interaction with hippocampal CA1 D2-dopamine receptors.

The cross-talk between dopaminergic and nitric oxide systems in the medial septal nucleus, and their distinct effects on anxiety-like behaviors in male rats.

Anxiety disorders, which have a noticeable global prevalence and may be caused by many factors, include a spectrum of disorders that share features of excessive fear- and anxiety-related behavioral disturbances. Different brain areas and neurotransmitter systems have been under investigation for anxiety-related disorders. In this study, we investigated the possible interaction between the dopaminergic and nitric oxide (NO) neurotransmitter systems in the medial septal nucleus and their roles in anxiety-like behaviors using elevated plus-maze (EPM) test in male rats. Our results showed that: (i) both D1-and D2-like receptor agonists, SKF-38393 and quinpirole, augmented anxiety-like behaviors at their two highest applied doses in the EPM test; (ii) both D1-and D2-like receptor antagonists, SCH- 23390 and sulpiride, reduced anxiety-like behaviors at their two highest applied doses in the EPM test; (iii) L-Arginine, a NO precursor, increased anxiety-like behaviors, but L-NAME, a non-specific nitric oxide synthase (NOS) inhibitor, reduced them in the EPM test; (iv) L-NAME could not reverse the anxiety-like parameters produced by SKF-38393, but it significantly reduced the anxiety-like behaviors induced by quinpirole; (v) Neither SCH- 23390 nor sulpiride changed anxiety-related behaviors induced by L-Arginine. It can be concluded that both dopaminergic and nitric oxide systems in the medial septal nucleus are involved in modulating anxiety-like behaviors. While NO has an involvement in the exerted effects by the D2-like agonist, such effects were not observed at the applied range of the doses for D1-and D2-like antagonists.

Blockade of orexin receptors in the hippocampal dentate gyrus reduced the extinction latency of morphine-induced place preference in male rats.

Relapse to drugs such as opioids is a major challenge in addiction therapy. It has been known that the orexinergic system has a significant role in mediating reward processing and addiction, as shown by the conditioned place preference (CPP). The dentate gyrus (DG) of the hippocampus receives orexinergic projections from the lateral hypothalamus that has been approved as a critical area arbitrating the maintenance of drug-seeking behavior following the extinction. The present study aimed to investigate the effects of intra-DG administration of the orexin-1 receptor (OX1R) and orexin-2 receptor (OX2R) antagonists on the extinction of morphine-induced CPP in male rats. Animals received different doses of SB334867 (as OX1R antagonist) or TCS OX2 29 (as OX2R antagonist) (0.5, 2.5, and 12.5 nM/0.5 µl DMSO 12 %) bilaterally into the DG during the extinction phase, after CPP had been induced by subcutaneous injection of morphine (5 mg/kg) during a 3-day conditioning phase. The conditioning scores were recorded during the test. The results demonstrated that intra-DG administration of the highest dose of OX1R antagonist (12.5 nM/0.5 µl DMSO 12 %) shortened the extinction latency of morphine-CPP compared to the DMSO group, while the OX2R antagonist did not significantly alter the latency. Findings imply that the blockade of OX1R, but not OX2R, within the DG facilitates the extinction of morphine-induced reward. In conclusion, the OX1R antagonist might be kept in mind as a convenient therapeutic factor in repressing drug-seeking behaviors in an optimum amount of treatment considering the low dose-treatments applied.

Cannabidiol attenuated the maintenance and reinstatement of extinguished methylphenidate-induced conditioned place preference in rats.

Methylphenidate (MPH) is a mild CNS stimulant that has been used in hyperactive children, and patients with neurodegenerative and major depressive disorders. Exposure to MPH-associated cues enhances craving and arousal in drug users. On the other hand, cannabidiol (CBD) has antipsychotic potential that might be useful in alleviating symptoms of drug addiction. The aim of this study was to investigate the effect of CBD administration on extinction and reinstatement of MPH-induced conditioning place preference (CPP) in rats. Male rats received MPH (1, 2.5 or 5 mg/kg, i.p) or morphine (5 or 10 mg/kg, s.c.) during the conditioning phase. Following the establishment of CPP, during extinction training, 60 min prior to every CPP session, animals were given daily ICV CBD (10 or 50 µg/5 µL), vehicle alone (DMSO) 10 % or were treatment-naïve. On the reinstatement day animals after receiving the initial dose of MPH, 0.5 mg/kg, and were placed into the CPP box to evaluate the CPP scoring for 10-min. Our findings indicated that morphine (5 and 10 mg/kg; s.c.) and MPH (1 and 2.5 mg/kg; i.p.) induced a CPP. The ICV administration of both doses of CBD (10 and 50 µg/5 µL) prevented the reinstatement of MPH-induced CPP, which displayed shorter extinction latency compared to treatment-naïve or DMSO 10 % groups. Therefore, CBD's site of action is a potential target for reducing the risk of MPH relapse; however, more investigation is required.

Role of orexinergic receptors in the dentate gyrus of the hippocampus in the acquisition and expression of morphine-induced conditioned place preference in rats.

Orexinergic projections derived from the lateral hypothalamus (LH) play a crucial role in the acquisition and expression of morphine-conditioned place preference (CPP). It has been demonstrated in previous that orexinergic receptors are expressed in the dentate gyrus (DG) region of the hippocampus, which receives projections of LH orexinergic neurons. This study examined the effects of intra-DG orexin-1 (OX1) and orexin-2 (OX2) receptor antagonists on the acquisition and expression of CPP induced by morphine. Two separate cannulas were inserted bilaterally into the DG, and a CPP paradigm was performed. The CPP scores and locomotor activities were recorded using Ethovision software. The results showed that intra-DG microinjection of SB334867 as a selective OX1R antagonist (0.5, 2.5, 12.5 nM/0.5 µl DMSO) or TCSOX229 as a selective OX2R antagonist (0.5, 2.5, 12.5 nM/0.5 µl DMSO) before a morphine subcutaneous injection (5 mg/kg) during a three-day conditioning phase dose-dependently represses the acquisition of morphine-induced CPP in rats. Furthermore, these antagonists reduced the CPP scores in the expression phase. Consequently, it was established that orexinergic receptors in the DG are involved in the acquisition and expression of morphine-induced CPP.

The blockade of D1- and D2-like dopamine receptors within the dentate gyrus attenuates food deprivation stress-induced reinstatement of morphine-extinguished conditioned place preference in rats.

The high rate of relapse to drug abuse is one of the main problems in the treatment of addiction. Stress plays an essential role in relapsing to drug abuse. The present study investigates the role of D1- and D2-like dopamine receptors in the dentate gyrus (DG) of the hippocampus on the reinstatement of morphine (5 mg/kg)-induced conditioned place preference (CPP) both by food deprivation stress (FDS) and a sub-threshold dose of morphine (0.5 mg/kg, s.c.). All the animals in this study experienced pre-test, conditioning, post-test (expression), extinction, and reinstatement phases. The CPP scores of the pre-test and post-test were compared between the groups, and a significant difference between the CPP scores of the pre- and post-test was the criterion for the induction of CPP. Extinction continued for each animal until the calculated score for two consecutive days became the same as the pre-test score. The animals received different doses of SCH-23390 or Sulpiride (0.25, 1 and 4 µg/0.5 µl vehicle), as D1- and D2-like dopamine receptor antagonists, into the DG. After the administration of the antagonists, the animals were deprived of food for 24 h. Then, on the reinstatement day, they received a sub-threshold dose of morphine and afterwards, the conditioning scores were measured. The results demonstrated that the effective doses 50% of SCH-23390 and Sulpiride on the reinstatement induced by FDS and morphine was 1.37 and 2.28 (µg/0.5 µl vehicle per side), respectively. The results also showed that both antagonists can lead to a decrease in morphine reinstatement, and this effect was in a dose-dependent manner. In conclusion, these results indicate that D1- and D2-like dopamine receptors in the DG may be a potential target for preventing relapse to drugs in stressful life conditions.

Involvement of orexin receptors within the hippocampal dentate gyrus in morphine-induced reinstatement in food-deprived rats.

The orexinergic system is found to cooperate in mediating stress-induced drug relapse. The orexinergic terminals innervate neurons of the hippocampal dentate gyrus (DG) which is a key structure in the maintenance and reinstatement of drug addiction. However, the specific contribution of intra-DG orexin receptors to stress-induced reinstatement has not been completely known. In the current study, the effects of intra-DG administration of SB334867, an orexin-1 receptor (OX1R) antagonist, and TCS OX2 29, an orexin-2 receptor (OX2R) antagonist, were investigated on the reinstatement induced by a sub-threshold dose of morphine and food deprivation (FD) stress. Adult male rats received different doses of SB334867 or TCS OX2 29 (3, 10, and 30 nM/0.5 µl DMSO 12%) bilaterally into the DG in separate groups, following the acquisition and extinction of morphine-induced conditioned place preference (CPP). Then, the reinstatement was evaluated by the 24 h FD stress and/or a sub-threshold dose of morphine (0.5 mg/kg, s.c.). CPP scores and locomotor activities were recorded during the test. The findings indicated that pre-treatment with the highest dose of SB334867 (30 nM) and two higher doses of TCS OX2 29 (10 and 30 nM) blocked the sub-threshold dose and FD stress-induced reinstatement of morphine. The effect of TCS OX2 29 on reduction of reinstatement was more pronounced than that of SB334867. It suggests a role for the orexin receptors, especially OX2R within the DG region in the stress-induced reinstatement of morphine-seeking behaviours in extinguished rats.