Locomotor Stimulation In Mice Research Articles

A high-affinity cocaine binding site associated with the brain acid soluble protein 1

Cocaine exerts its stimulant effect by inhibiting dopamine (DA) reuptake, leading to increased dopamine signaling. This action is thought to reflect the binding of cocaine to the dopamine transporter (DAT) to inhibit its function. However, cocaine is a relatively weak inhibitor of DAT, and many DAT inhibitors do not share cocaine’s behavioral actions. Further, recent reports show more potent actions of the drug, implying the existence of a high-affinity receptor for cocaine. We now report high-affinity binding of cocaine associated with the brain acid soluble protein 1 (BASP1) with a dissociation constant (Kd) of 7 nM. Knocking down BASP1 in the striatum inhibits [3H]cocaine binding to striatal synaptosomes. Depleting BASP1 in the nucleus accumbens but not the dorsal striatum diminishes locomotor stimulation in mice. Our findings imply that BASP1 is a pharmacologically relevant receptor for cocaine.

Role of Efficacy as a Determinant of Locomotor Activation Induced by Mu Opioid Receptor Ligands in Male and Female Mice

Background Ligand efficacy is a key determinant of drug effects mediated by receptor targets, and efficacy can be exploited to generate safer therapeutics. As one step in a broader project to examine efficacy requirements for different mu opioid receptors (MOR) effects, this study evaluated locomotor stimulation in mice produced by 8 MOR ligands that vary from low to high efficacy. Additionally, opioid agonist/antagonist mixtures provide a novel strategy to precisely calibrate levels of MOR activation, measure efficacy requirements for different MOR-mediated endpoints, and identify efficacy levels that might optimally balance effectiveness to produce therapeutic effects while minimizing side effects. Accordingly, effects of MOR ligands were compared to effects of fixed-proportion mixtures of the opioid agonist fentanyl and antagonist naltrexone. We hypothesized that MOR ligands would produce an efficacy-dependent increase in locomotor stimulation, and that fentanyl/naltrexone mixtures could be used to compare efficacy requirements for mouse locomotor stimulation relative to other in vivo and in vitro opioid effects. Methods We determined locomotor activity in adult male and female ICR mice after acute treatment with (in order of increasing efficacy) naltrexone, NAQ, nalbuphine, buprenorphine, hydrocodone, morphine, fentanyl, and methadone. Fentanyl/naltrexone mixtures were studied in fixed proportions of 1:1, 3.2:1, 10:1, 32:1, and 100:1. Dose-effect curves for each drug or mixture were determined in separate groups of 12 mice (6 male, 6 female). Treatments were administered subcutaneously 5 min before 100-min sessions, and locomotor activity was quantified as photobeam breaks in chambers with a floor area of 16.8 × 12.7 cm2. Data were analyzed by one- or two-way ANOVA, and the criterion for significance was p<0.05. Additionally, data with the mixtures were used to compare efficacy requirements for locomotor stimulation in this study relative to previously published data for other in vitro and in vivo effects. Results MOR ligands produced an efficacy-dependent increase in maximal locomotor stimulation. Similarly, for the fentanyl/naltrexone mixtures, increasing fentanyl proportions (pFentanyl) also produced graded increases in maximum stimulation (Emax). The curve relating pFentanyl to Emax was used to quantify the effective pFentanyl required to produce 50% of the fentanyl-alone Emax (EP50), and the EP50 (95%CL) served as a metric of the efficacy requirement for any opioid endpoint. In this case, the EP50 for locomotor stimulation in mice was high relative to EP50 values for other in vivo endpoints including fentanyl discrimination in rats and thermal antinociception in mice, rats, and rhesus monkeys, but lower than the EP50 for an in vitro assay of GTPɣS binding in mMOR-CHO cells. Conclusions These results confirm efficacy-dependent stimulation of locomotor activity by MOR ligands in mice. Additionally, these results illustrate the utility of fentanyl/naltrexone mixtures to precisely calibrate net MOR activation, quantify efficacy requirements across different opioid effects, and identify efficacy levels that might be sufficient to produce some drug effects but not others.

Novel Synthetic Cathinone Eutylone and its Structural Isomers Interact with Monoamine Transporters and Induce Locomotor Stimulation in Mice

Synthetic cathinones are a class of new psychoactive substances that produce psychostimulant effects and pose public health risks. Currently, derivatives of the synthetic cathinone, methylone, are being confiscated worldwide. Eutylone is a popular emerging methylone analog with ethyl substitutions at the a-carbon and amine positions. Little information is presently available about the pharmacological effects of eutylone. Therefore, based on its structure we hypothesized that the compound interacts with monoamine transporters in the brain and produces locomotor stimulation akin to other cathinone psychostimulants. To test this hypothesis, we compared the effects of eutylone and its structural isomers, pentylone and dibutylone, using in vitro transporter assays in rat brain synaptosomes and in vivo locomotor activity assessments in mice. All drugs displayed dose-dependent inhibition of [3H]neurotransmitter uptake at dopamine transporters (DAT) and norepinephrine transporters (NET), with 10-fold more potent effects at DAT (IC50 = ~120 nM). Contrary to dibutylone, which displayed no effects at serotonin transporters (SERT), eutylone and pentylone inhibited uptake at SERT (0.69 & 1.24 μM respectively) and displayed weak partial releasing actions at SERT (both ~50% of maximal response). All drugs stimulated dose-dependent locomotion in mice, with eutylone displaying the most potent and efficacious effects (ED50 = 2.1 mg/kg, s.c.). Rank order of ED50 potency values for locomotor stimulation was eutylone ≥ pentylone > cocaine > dibutylone. Our results demonstrate that eutylone is a potent monoamine transporter inhibitor that displays preference for DAT and induces locomotor stimulation. Eutylone displayed pharmacological effects very similar to those produced by pentylone, which suggests that eutylone will exhibit similar abuse liability and pose risks for psychostimulant side-effects in human users.

A ghrelin receptor antagonist reduces the ability of ghrelin, alcohol or amphetamine to induce a dopamine release in the ventral tegmental area and in nucleus accumbens shell in rats

The orexigenic peptide ghrelin increases the release of dopamine in the nucleus accumbens (NAc) shell via central ghrelin receptors, especially those located in the ventral tegmental area (VTA). The activity of the VTA dopamine neurons projecting to NAc shell, involves somatodendritic dopamine release within the VTA. However, the effects of ghrelin on the concomitant dopamine release in the VTA and NAc shell is unknown. It is further unknown whether addictive drugs, such as alcohol and amphetamine, enhance the dopamine levels in both these areas via ghrelin receptor dependent mechanisms. Thus, the effects of a ghrelin receptor antagonist, JMV2959, on the ability of i) central ghrelin ii) systemic alcohol or iii) systemic amphetamine to increase the dopamine release in the VTA and in the NAc shell in rats by using in vivo microdialysis was explored. We showed that systemic administration of JMV2959 blocks the ability of central ghrelin to increases dopamine release in the VTA and the NAc shell, and reduces the alcohol- and amphetamine-induced dopamine release in both these areas. Locomotor activity studies was then conducted in an attempt to correlate the ghrelin-induced dopamine release in the VTA to a behavioural outcome. These revealed that local infusion of a dopamine D1 receptor antagonist into the VTA blocks the ability of central ghrelin to cause a locomotor stimulation in mice. Collectively, this study adds to the growing body of evidence indicating that ghrelin signalling modulates the ability of ghrelin, and addictive drugs, to activate the mesoaccumbal dopamine pathway.

Activation of the amylin pathway modulates cocaine-induced activation of the mesolimbic dopamine system in male mice

Besides food intake reduction, activation of the amylin pathway by salmon calcitonin (sCT), an amylin and calcitonin receptor agonist, inhibits alcohol-mediated behaviors in rodents. This involves brain areas processing reward, i.e. the laterodorsal (LDTg), ventral tegmental area (VTA) and nucleus accumbens (NAc). However, the effects of stimulation of the amylin pathway on behaviors caused by cocaine and the brain areas involved in these processes have not yet been investigated. We therefore explored in male mice, the effects of systemic administration of sCT on cocaine-induced locomotor stimulation, dopamine release in the NAc and cocaine reward, as well as reward-dependent memory of cocaine, in the conditioned place preference (CPP) paradigm. Moreover, the outcome of systemic sCT and cocaine co-administration for five days on locomotor activity was investigated. Lastly, the impact of sCT infusions into the LDTg, VTA, NAc shell or core on cocaine-evoked locomotor stimulation was explored. We found that sCT attenuated cocaine-induced locomotor stimulation and accumbal dopamine release, without altering cocaine's rewarding properties or reward-dependent memory retrieval in the CPP paradigm. Five days of cocaine administration caused locomotor stimulation in mice pre-treated with vehicle, but not with sCT. In mice infused with vehicle into the aforementioned reward-related areas, cocaine caused locomotor stimulation, a response that was not evident following sCT infusions. The current findings suggest a novel role for the amylinergic pathway as regulator of cocaine-evoked activation of the mesolimbic dopamine system, opening the way for the investigation of the amylin signalling in the modulation of other drugs of abuse.

Effects of sub-chronic amylin receptor activation on alcohol-induced locomotor stimulation and monoamine levels in mice

RationaleAmylin receptors consist of the calcitonin receptor (CTR) and one of three receptor activity-modifying proteins (RAMPs). The identification of amylin receptors in areas processing reward, namely laterodorsal tegmental area (LDTg), ventral tegmental area (VTA), and nucleus accumbens (NAc), has attributed them a role as reward regulators. Indeed, acute activation of amylin receptors by the amylin receptor agonist salmon calcitonin (sCT) attenuates alcohol-induced behaviours in rodents.ObjectivesThe effects of long-term administration of sCT on alcohol-related behaviours and the molecular mechanisms underlying these processes are not yet elucidated. To fill this knowledge gap, we investigated the effects of sub-chronic sCT treatment on the locomotor stimulatory responses to alcohol in mice and the molecular pathways involved.MethodsWe assessed the behavioural effects of sub-chronic sCT treatment by means of locomotor activity experiments in mice. We used western blot to identify changes of the CTR levels and ex vivo biochemical analysis to detect changes in monoamines and their metabolites.ResultsAfter discontinuation for 5 days of sCT treatment, alcohol did not induce locomotor stimulation in mice pre-treated with sCT when compared with vehicle, without altering secondary behavioural parameters of the locomotor activity experiment or the protein levels of the CTR in reward-related areas in the same set of animals. Moreover, repeated sCT treatment altered monoaminergic neurotransmission in various brain areas, including increased serotonin and decreased dopamine turnover in the VTA. Lastly, we identified a differential effect of repeated sCT and acute alcohol administration on alcohol-induced locomotion in mice, where sCT initially attenuated and later increased this alcohol response. It was further found that this treatment combination did not affect secondary behavioural parameters measured in this locomotor activity experiments.ConclusionsThese data suggest that sub-chronic sCT treatment differentially alters the ability of alcohol to cause locomotor stimulation, possibly through molecular mechanisms involving various neurotransmitter systems and not the CTR levels per se.

Glucagon-like peptide-1 receptors within the nucleus of the solitary tract regulate alcohol-mediated behaviors in rodents

The ability of glucagon-like peptide-1 (GLP-1) to reduce food intake involves activation of GLP-1 receptors (GLP-1R) in the nucleus of the solitary tract (NTS). It has also been demonstrated that systemic administration of GLP-1R agonists attenuates alcohol-mediated behaviors via, to date, unknown mechanisms. Therefore, we evaluated the effects of NTS-GLP-1R activation by exendin-4 (Ex4) on alcohol-induced locomotor stimulation, accumbal dopamine release and memory of alcohol reward in the conditioned place preference (CPP) model in mice. Moreover, the ability of Ex4 infusion into the NTS on alcohol intake was explored in rats. Ex4 into the NTS inhibits the acute effects of alcohol as measured by alcohol-induced locomotor stimulation, accumbal dopamine release and the memory consolidation of alcohol reward in the CPP paradigm. In addition, NTS-Ex4 dose-dependently decreases alcohol intake in rats consuming alcohol for 12 weeks. Pharmacological suppression of GLP-1R in the NTS prevents the ability of systemic Ex4 to block the alcohol-induced locomotor stimulation in mice. These data add a functional role of GLP-1R within the NTS, involving alcohol-related behaviors. In addition, they may provide insight into the GLP-1R containing brain areas that modulate the ability of GLP-1R agonists to reduce alcohol reinforcement. Collectively, this further supports GLP-1R as potential treatment targets for alcohol use disorder.

Role of monoaminergic systems and ambient temperature in bath salts constituent 3,4-methylenedioxypyrovalerone (MDPV)-elicited hyperthermia and locomotor stimulation in mice

3,4-Methylenedioxypyrovalerone (MDPV) is a common constituent of illicit bath salts products, and in vitro studies implicate monoamine transporters as mediators of its pharmacological effects. Locomotor and thermoregulatory effects of MDPV depend on ambient temperature, so the current studies aimed to gauge the involvement of dopamine (DA), norepinephrine (NE), and serotonin (5-HT) in MDPV-induced locomotor stimulation and hyperthermia in the mouse at different ambient temperatures. Mice were pretreated with the selective 5-HT-reuptake inhibitor fluoxetine (3 mg/kg), the NE-reuptake inhibitor desipramine (3 mg/kg), the DA-reuptake inhibitor bupropion (10 mg/kg), or saline, followed by 10 mg/kg MDPV while thermoregulation and locomotor activity were monitored via radiotelemetry. In other studies, mice were pretreated for three days with saline, 100 mg/kg of the tryptophan hydroxylase inhibitor para-chlorophenylalanine (p-CPA), or 100 mg/kg of the tyrosine hydroxylase inhibitor α-methyl-para-tyrosine (α-MPT) before receiving 10 mg/kg MDPV on the fourth day. All manipulations were conducted at both 20 °C and 28 °C ambient temperatures. MDPV increased locomotor activity under both ambient conditions and modestly increased core body temperature at 20 °C; however, neither pretreatment with monoamine reuptake inhibitors nor monoamine synthesis inhibitors significantly altered these effects. At 28 °C, MDPV induced a more pronounced hyperthermic effect which was attenuated by bupropion, desipramine, or fluoxetine pretreatment, but not by the monoamine synthesis inhibitors. These results suggest that MDPV may have a more complex pharmacological profile than suggested by in vitro studies, perhaps extending beyond interactions with monoamine transporters. A more thorough binding profile of MDPV at various brain recognition sites should be developed.This article is part of the Special Issue entitled ‘Designer Drugs and Legal Highs.’

Novel and High Affinity 2-[(Diphenylmethyl)sulfinyl]acetamide (Modafinil) Analogues as Atypical Dopamine Transporter Inhibitors.

The development of pharmacotherapeutic treatments of psychostimulant abuse has remained a challenge, despite significant efforts made toward relevant mechanistic targets, such as the dopamine transporter (DAT). The atypical DAT inhibitors have received attention due to their promising pharmacological profiles in animal models of cocaine and methamphetamine abuse. Herein, we report a series of modafinil analogues that have an atypical DAT inhibitor profile. We extended SAR by chemically manipulating the oxidation states of the sulfoxide and the amide functional groups, halogenating the phenyl rings, and/or functionalizing the terminal nitrogen with substituted piperazines, resulting in several novel leads such as 11b, which demonstrated high DAT affinity (Ki = 2.5 nM) and selectivity without producing concomitant locomotor stimulation in mice, as compared to cocaine. These results are consistent with an atypical DAT inhibitor profile and suggest that 11b may be a potential lead for development as a psychostimulant abuse medication.

The Anorexigenic Peptide Neuromedin U (NMU) Attenuates Amphetamine-Induced Locomotor Stimulation, Accumbal Dopamine Release and Expression of Conditioned Place Preference in Mice.

Amphetamine dependence, besides its substantial economical consequence, is a serious cause of mortality and morbidity. By investigations of the neurochemical correlates through which addictive drugs, such as amphetamine, activate the mesoaccumbal dopamine system unique targets for treatment of drug addiction can be identified. This reward link consists of a dopamine projection from the ventral tegmental area to the nucleus accumbens (NAc) suggesting that these brain areas are important for reward. The physiological function of gut-brain peptides has expanded beyond food intake modulation and involves regulation of drug reinforcement. A novel candidate for reward regulation is the anorexigenic peptide neuromedin U (NMU). We therefore investigated the effects of intracerebroventricular (icv) administration of NMU on amphetamine’s well-documented effects on the mesoaccumbal dopamine system, i.e. locomotor stimulation and accumbal dopamine release in mice. In addition, the effect of accumbal NMU administration on locomotor activity was examined. The effect of NMU, icv or intra-NAc, on the expression of conditioned place preference (CPP) was elucidated. Firstly, we showed that icv administration of NMU attenuate the amphetamine-induced locomotor stimulation, accumbal dopamine release and expression of CPP in mice. Secondly, we found that a lower dose of NMU (icv) reduce the amphetamine-induced locomotor stimulation in mice. Thirdly, we demonstrated that NMU administration into the NAc block the ability of amphetamine to cause a locomotor stimulation in mice. However, accumbal NMU administration did not attenuate the amphetamine-induced expression of CPP in mice. Our novel data suggest that central NMU signalling is involved in development of amphetamine dependence.

Sub-chronic Ghrelin Receptor Blockade Attenuates Alcohol- and Amphetamine-Induced Locomotor Stimulation in Mice.

Ghrelin initially emerged as a gut-brain hormone controlling food intake, meal initiation and appetite mainly via hypothalamic circuits in both rodents and humans. The findings that ghrelin receptors (GHS-R1A) are expressed in reward-related areas, including the nucleus accumbens (NAc) and ventral tegmental area (VTA), suggest that ghrelin is a novel reward regulator. Indeed, ghrelin signalling mediates the rewarding and motivational properties of addictive drugs. In addition, daily co-administration of a GHS-R1A antagonist and various addictive drugs prevents the drug-induced locomotor sensitization in rats. The present series of experiment were designed to evaluate the effect of repeated pharmacological GHS-R1A suppression on drug-induced locomotor stimulation in more detail. We showed that sub-chronic pre-treatment of the GHS-R1A antagonist, JMV2959, attenuated the ability of acute administration of alcohol as well as of amphetamine to stimulate locomotion. However, there was no effect of sub-chronic JMV2959 treatment on locomotor activity per se or on the expression of the GHS-R1A gene (Ghsr) in the VTA or the NAc compared with vehicle treatment. In addition, sub-chronic ghrelin treatment caused a locomotor sensitization. While previous research has pinpointed ghrelin as an appetite regulator the present study together with previous studies suggest that ghrelin signalling modulates various reward-mediated behaviours in rodents. Collectively, this suggests that the GHS-R1A could be a key target for novel treatment strategies for addiction.

Adenosinergic regulation of binge-like ethanol drinking and associated locomotor effects in male C57BL/6J mice

We recently observed that the addition of caffeine (a nonselective adenosine receptor antagonist) to a 20% ethanol solution significantly altered the intoxication profile of male C57BL/6J (B6) mice induced by voluntary binge-like consumption in the ‘Drinking-in-the-Dark’ (DID) paradigm. In the current study, the roles of A1 and A2A adenosine receptor subtypes, specifically, in binge-like ethanol consumption and associated locomotor effects were explored. Adult male B6 mice (PND 60–70) were allowed to consume 20% ethanol (v/v) or 2% sucrose (w/v) for 6days via DID. On day 7, mice received a systemic administration (i.p.) of the A1 antagonist DPCPX (1, 3, 6mg/kg), the A2A antagonist MSX-3 (1, 2, 4mg/kg), or vehicle immediately prior to fluid access in DID. Antagonism of the A1 receptor via DPCPX was found to dose-dependently decrease binge-like ethanol intake and associated blood ethanol concentrations (p's<0.05), although no effect was observed on sucrose intake. Antagonism of A2A had no effect on ethanol or sucrose consumption, however, MSX-3 elicited robust locomotor stimulation in mice consuming either solution (p's<0.05). Together, these findings suggest unique roles for the A1 and A2A adenosine receptor subtypes in binge-like ethanol intake and its associated locomotor effects.

Activation of the GABAB Receptor Prevents Nicotine-Induced Locomotor Stimulation in Mice

Recent studies demonstrated that activation of the GABAB receptor, either by means of orthosteric agonists or positive allosteric modulators (PAMs), inhibited different nicotine-related behaviors, including intravenous self-administration and conditioned place preference, in rodents. The present study investigated whether the anti-nicotine effects of the GABAB receptor agonist, baclofen, and GABAB PAMs, CGP7930, and GS39783, extend to nicotine stimulant effects. To this end, CD1 mice were initially treated with baclofen (0, 1.25, and 2.5 mg/kg, i.p.), CGP7930 (0, 25, and 50 mg/kg, i.g.), or GS39783 (0, 25, and 50 mg/kg, i.g.), then treated with nicotine (0 and 0.05 mg/kg, s.c.), and finally exposed to an automated apparatus for recording of locomotor activity. Pretreatment with doses of baclofen, CGP7930, or GS39783 that did not alter locomotor activity when given with nicotine vehicle fully prevented hyperlocomotion induced by 0.05 mg/kg nicotine. These data extend to nicotine stimulant effects the capacity of baclofen and GABAB PAMs to block the reinforcing, motivational, and rewarding properties of nicotine. These data strengthen the hypothesis that activation of the GABAB receptor may represent a potentially useful, anti-smoking therapeutic strategy.

The antipsychotic aripiprazole antagonizes the ethanol- and amphetamine-induced locomotor stimulation in mice

Most drugs of abuse cause a locomotor stimulation, an effect, at least in part, mediated by increased accumbal dopamine (DA) overflow. Locomotor stimulation has been suggested to be a putative endophenotype for drug addiction. We therefore investigated the effects of aripiprazole, a partial DA D 2-receptor agonist, on ethanol as well as amphetamine-induced locomotor stimulation. In the present series of experiments, we found that aripiprazole (1.25 mg/kg, intraperitoneally [i.p.]) antagonized ethanol (1.75 g/kg, i.p.) as well as amphetamine (2 mg/kg, i.p.)induced locomotor stimulation in mice. We suggest that this effect might be related to aripiprazole's ability to alleviate drug-induced hyperdopaminergia without causing hypodopaminergia. Given that altered DA functions in drug dependence have been observed, it may be suggested that aripiprazole could be a new treatment strategy for treatment of drug dependence.

Habituation to test procedure modulates the involvement of dopamine D2- but not D1-receptors in ethanol-induced locomotor stimulation in mice

Novelty associated with behavioral testing has been shown to enhance psychostimulant- and morphine-induced locomotor stimulation. Evidence has demonstrated that novelty increases dopamine (DA) activity, and habituation to a novel environment reduces such activation. However, it is not clear whether novelty modulates ethanol-induced behavioral stimulation and whether DA plays a role in this effect. The present work sought to demonstrate a role of habituation to test procedure as a factor that could modulate the involvement of DA in ethanol-induced locomotor stimulation. Non-habituated (NH) and habituated (H) Swiss mice pretreated with DA D1- (SCH23390; 0-0.045 mg/kg) or D2-receptor (sulpiride; 0-50 mg/kg) antagonists were tested for ethanol (0-2.5 g/kg)-induced locomotor stimulation. Experiments with amphetamine (0-4 mg/kg), morphine (0-5 mg/kg) and caffeine (0-15 mg/kg)were designed to compare their results to those obtained with ethanol. The effect of the non-selective opioid receptor antagonist naltrexone (0-1.5 mg/kg) was also tested on ethanol-induced locomotor stimulation. NH and H animals did not differ in their locomotor response to ethanol or caffeine; however, amphetamine- and morphine-induced stimulation was greater in NH than in H mice. SCH23390 only reduced ethanol-induced stimulation at doses that also reduced spontaneous activity in both NH and H mice. Sulpiride decreased ethanol-stimulated behavior only in the NH condition. Habituation did not modify the effect of sulpiride on amphetamine-, morphine- or caffeine-induced activation. Naltrexone (0-1.5 mg/kg) reduced ethanol-induced stimulation regardless of habituation. The present data suggest that the participation of DA D2-receptors in ethanol-induced behavioral stimulation requires the presence of novelty. Results also support the involvement of neurotransmitter systems other than DA (i.e., endogenous opioid system) as important substrates mediating ethanol-induced locomotor activation.

Combination of adenosine A1 and A2A receptor blocking agents induces caffeine-like locomotor stimulation in mice

The spontaneous locomotor activity of C57BL/6J mice was examined, using an automated detection system based on infra-red beams, after administration of caffeine (3–30 mg/kg, i.p.), the adenosine A2A receptor selective antagonist SCH 58261 (0.312–2.5 mg/kg, i.p.) and the A1 selective antagonist DPCPX (1.25–5 mg/kg, i.p.). SCH 58261 failed to influence motor activity in mice habituated to the test environment. DPCPX produced a small increase in motility and locomotion (significant at the dose of 5.0 mg/kg), much weaker than that produced by caffeine. Combined administration of DPCPX (1.2 mg/kg, i.p.) and SCH 58261 (1.2 mg/kg, i.p.) produced stimulation of motility and locomotion comparable with the effect of caffeine (15 mg/kg, i.p.). In contrast to motility and locomotion, rearing counts were not significantly influenced by DPCPX, SCH 58261, their combination, or by caffeine. Caffeine (15 mg/kg, i.p.) caused an increase in NGFI-A mRNA (an immediate early gene was chosen as an index of neuronal activation) in the piriform cortex 4 h after injection. This effect was reproduced by the combination of A1 and A2A receptor antagonist. It is hypothesised that the stimulatory effect of low doses of caffeine in C57BL/6J mice is due to concomitant blockade of both A1 and A2A adenosine receptors.

GABA B receptor inhibition causes locomotor stimulation in mice

The present study investigated the effect of the administration of the GABA B receptor antagonists, SCH 50911 [(2 S)(+)-5,5-dimethyl-2-morpholineacetic acid], CGP 46381 [(3-aminopropyl)(cyclohexylmethyl)phosphinic acid] and CGP 52432 (3-[[(3,4-dichlorophenyl)methyl]amino]propyl]diethoxymethyl)phosphinic acid), on spontaneous locomotor activity in mice. All drugs were acutely administered at the doses of 10 and 30 mg/kg (i.p.). The dose of 30 mg/kg of all drugs resulted in a significant stimulation of locomotor activity. The locomotor stimulation elicited by SCH 50911 was completely blocked by haloperidol (0.1 mg/kg, i.p.), suggesting that hyperactivity induced by blockade of the GABA B receptor is mediated by enhanced dopamine release. These results suggest the existence of a GABA B receptor-mediated tonic inhibition of dopamine neurons.

Acetylcodeine, an impurity of illicitly manufactured heroin, elicits convulsions, antinociception, and locomotor stimulation in mice

Acetylcodeine is one of the major impurities present in illicitly manufactured heroin (diacetylmorphine). Data on its pharmacology and toxicology are limited and its ability to alter the toxic effects of diacetylmorphine is not known. The first objective of the present study was to compare the acute pharmacological and toxicological effects of acetylcodeine to those of codeine and diacetylmorphine in mice by assessing nociception in the tail-flick test, locomotor stimulation, and convulsive behavior. The second goal of this study was to determine whether acetylcodeine would alter the convulsant effects of diacetylmorphine. The antinociceptive potencies of acetylcodeine and codeine were similar, as reflected by their ED 50 (95% confidence limits) values of 35 (29–44) and 51 (40–65) μmol/kg, respectively. Acetylcodeine was somewhat less potent than codeine in stimulating locomotor behavior, with ED 50 values of 28 (22–37) and 12 (6–24) μmol/kg, respectively. Diacetylmorphine was considerably more potent than the other two drugs, producing antinociception and locomotor stimulation at ED 50 values of 2.4 (1.4–4.1) and 0.65 (0.36–1.2) μmol/kg, respectively. On the other hand, the convulsant effects of acetylcodeine (ED 50=138 (121–157) μmol/kg) and diacetylmorphine (ED 50=115 (81–163) μmol/kg) were similar in potency and both were more potent than codeine (ED 50=231 (188–283) μmol/kg). Finally, a subthreshold dose of acetylcodeine (72 μmol/kg) decreased the convulsant ED 50 dose of diacetylmorphine to 40 (32–49). These findings suggest that the convulsant effects of acetylcodeine are more potent than predicted by its effects on locomotor activity and antinociception. The observation that acetylcodeine potentiated the convulsant effects of diacetylmorphine suggests a mechanism for some of the heroin-related deaths reported in human addicts.

Structure-activity relationships at monoamine transporters and muscarinic receptors for N-substituted-3alpha-(3'-chloro-, 4'-chloro-, and 4',4''-dichloro-substituted-diphenyl)methoxytropanes.

The design, synthesis, and evaluation of 3alpha-(diphenylmethoxy)tropane (benztropine) analogues have provided potent and selective probes for the dopamine transporter. Structure-activity relationships (SARs) have been developed that contrast with those described for cocaine, despite significant structural similarity. Furthermore, behavioral evaluation of many of the benztropine analogues in animal models of cocaine abuse has suggested that these two classes of tropane-based dopamine uptake inhibitors have distinct pharmacological profiles. In general, the benztropine analogues do not demonstrate efficacious locomotor stimulation in mice, do not fully substitute for a cocaine discriminative stimulus, and are not appreciably self-administered in rhesus monkeys. These compounds are generally more potent than cocaine as dopamine uptake inhibitors in vitro, although their actions in vivo are not consistent with this action. These observations suggest that differing binding profiles at the serotonin and norepinephrine transporters as well as at muscarinic receptors might have significant impact on the pharmacological actions of these compounds. In addition, by varying the structures of the parent compounds and thereby modifying their physical properties, pharmacokinetics as well as pharmacodynamics will be directly affected. Therefore, in an attempt to systematically evaluate the impact of chemical modification on these actions, a series of N-substituted (H, CH3, allyl, benzyl, propylphenyl, and butylphenyl) analogues of 3'-chloro-, 4'-chloro-, and 4,4''-dichloro-3alpha-(diphenylmethoxy)tropanes were synthesized. These compounds were evaluated for displacement, in rat tissue, of [3H]WIN 35,428 from the dopamine transporter, [3H]citalopram from the serotonin transporter, [3H]nisoxetine from the norepinephrine transporter, and [3H]pirenzepine from muscarinic m1 receptors. SARs were developed and compared to a series of N-substituted-3alpha-(bis-4'-fluorophenyl)methoxytropanes. The present SARs followed previously reported studies with the single exception of the N-butylphenyl substituent, which did not provide the high affinity binding in any of these three sets of analogues, as it did in the 4',4''-difluoro series. X-ray crystallographic analyses of the three parent ligands (1a, 2a, and 3a) were compared to that of 3alpha-(bis-4'-fluorophenyl)methoxytropane which provided supportive evidence toward the proposal that the combination of steric bulk in both the 3-position and the N-substituent, in this class of compounds, is not optimal for binding at the dopamine transporter. These studies provide binding profile data that can now be used to correlate with future behavioral analyses of these compounds and may provide insight into the kind of binding profile that might be targeted as a potential treatment for cocaine abuse.

T-type calcium channel blockade blocks bromocriptine-induced locomotor stimulation in mice

T-type calcium channel blockade blocks bromocriptine-induced locomotor stimulation in mice