ABSTRACT
The dynorphin/κ-opioid receptor system has been implicated in the pathogenesis and pathophysiology of several psychiatric disorders. In the present review, we present evidence indicating a key role for this system in modulating neurotransmission in brain circuits that subserve mood, motivation, and cognitive function. We overview the pharmacology, signaling, post-translational, post-transcriptional, transcriptional, epigenetic and cis regulation of the dynorphin/κ-opioid receptor system, and critically review functional neuroanatomical, neurochemical, and pharmacological evidence, suggesting that alterations in this system may contribute to affective disorders, drug addiction, and schizophrenia. We also overview the dynorphin/κ-opioid receptor system in the genetics of psychiatric disorders and discuss implications of the reviewed material for therapeutics development.
Subject(s)
Dynorphins/physiology , Mental Disorders/etiology , Receptors, Opioid, kappa/physiology , Brain/physiology , Central Nervous System Stimulants/pharmacology , Cyclic AMP Response Element-Binding Protein/physiology , Dynorphins/genetics , Enkephalins/genetics , Epigenesis, Genetic , Gene Expression Regulation , Humans , Mental Disorders/drug therapy , Models, Animal , Protein Precursors/genetics , Receptors, Opioid, kappa/agonists , Receptors, Opioid, kappa/genetics , Self Stimulation , Signal TransductionABSTRACT
Mu opioid receptor (MOR) regulation of somatodendritic dopamine neurotransmission in the ventral tegmental area (VTA) was investigated using conventional microdialysis in freely moving rats and mice. Reverse dialysis of the MOR agonist DAMGO (50 and 100 microm) into the VTA of rats produced a concentration-dependent increase in dialysate dopamine concentrations. Basal dopamine overflow in the VTA was unaltered in mice lacking the MOR gene. However, basal gamma-aminobutyric acid (GABA) overflow in these animals was significantly increased, whereas glutamate overflow was decreased. Intra-VTA perfusion of DAMGO into wild-type (WT) mice increased dopamine overflow. GABA concentrations were decreased, whereas glutamate concentrations in the VTA were unaltered. Consistent with the loss of MOR, no effect of DAMGO was observed in MOR knockout (KO) mice. These data provide the first direct demonstration of tonically active MOR systems in the VTA that regulate basal glutamatergic and GABAergic neurotransmission in this region. We hypothesize that increased GABAergic neurotransmission following constitutive deletion of MOR is due to the elimination of a tonic inhibitory influence of MOR on GABAergic neurons in the VTA, whereas decreased glutamatergic neurotransmission in MOR KO mice is a consequence of intensified GABA tone on glutamatergic neurons and/or terminals. As a consequence, somatodendritic dopamine release is unaltered. Furthermore, MOR KO mice do not exhibit the positive correlation between basal dopamine levels and the glutamate/GABA ratio observed in WT mice. Together, our findings indicate a critical role of VTA MOR in maintaining an intricate balance between excitatory and inhibitory inputs to dopaminergic neurons.
Subject(s)
Dendrites/physiology , Dopamine/metabolism , Glutamic Acid/physiology , Receptors, Opioid, mu/physiology , gamma-Aminobutyric Acid/physiology , Animals , Dendrites/drug effects , Enkephalin, Ala(2)-MePhe(4)-Gly(5)-/pharmacology , Male , Mice , Mice, Inbred C57BL , Mice, Knockout , Rats , Rats, Sprague-Dawley , Receptors, Opioid, mu/agonists , Somatosensory Cortex/drug effects , Somatosensory Cortex/physiology , Ventral Tegmental Area/drug effects , Ventral Tegmental Area/physiologyABSTRACT
Studies in experimental animals have shown that individuals exhibiting enhanced sensitivity to the locomotor-activating and rewarding properties of drugs of abuse are at increased risk for the development of compulsive drug-seeking behavior. The purpose of the present study was to assess the effect of constitutive deletion of delta-opioid receptors (DOPr) on the rewarding properties of morphine as well as on the development of sensitization and tolerance to the locomotor-activating effects of morphine. Locomotor activity testing revealed that mice lacking DOPr exhibit an augmentation of context-dependent sensitization following repeated, alternate injections of morphine (20 mg/kg; s.c.; 5 days). In contrast, the development of tolerance to the locomotor-activating effects of morphine following chronic morphine administration (morphine pellet: 25 mg: 3 days) is reduced relative to WT mice. The conditioned rewarding effects of morphine were reduced significantly in DOPrKO mice as compared to WT controls. Similar findings were obtained in response to pharmacological inactivation of DOPr in WT mice, indicating that observed effects are not due to developmental adaptations that occur as a consequence of constitutive deletion of DOPr. Together, these findings indicate that the endogenous DOPr system is recruited in response to both repeated and chronic morphine administration and that this recruitment serves an essential function in the development of tolerance, behavioral sensitization, and the conditioning of opiate reward. Importantly, they demonstrate that DOPr has a distinct role in the development of each of these drug-induced adaptations. The anti-rewarding and tolerance-reducing properties of DOPr antagonists may offer new opportunities for the treatment and prevention of opioid dependence as well as for the development of effective analgesics with reduced abuse liability.
Subject(s)
Drug Tolerance/physiology , Morphine/pharmacology , Motor Activity/drug effects , Receptors, Opioid, delta/metabolism , Reward , Animals , Conditioning, Psychological/physiology , Male , Mice , Mice, Inbred C57BL , Mice, Knockout , Naltrexone/analogs & derivatives , Naltrexone/pharmacology , Narcotic Antagonists/pharmacology , Receptors, Opioid, delta/antagonists & inhibitors , Spatial Behavior/drug effectsABSTRACT
Drug addiction is a chronic, relapsing disorder that is characterized by a compulsion to take drug regardless of the adverse consequences that may ensue. Although the involvement of mesoaccumbal dopamine neurons in the initiation of drug abuse is well-established, neuroadaptations within the limbic cortical- striatopallidal circuit that occur as a consequence of repeated drug use are thought to lead to the behavioral dysregulation that characterizes addiction. Opioid receptors and their endogenous ligands are enriched in brain regions comprising this system and are, thus, strategically located to modulate neurotransmission therein. This article will review data suggesting an important role of mu-opioid receptor (MOPr) and delta opioid receptor (DOPr) systems in mediating the rewarding effects of several classes of abused drugs and that aberrant activity of these opioid systems may not only contribute to the behavioral dysregulation that characterizes addiction but to individual differences in addiction vulnerability.
Subject(s)
Brain/metabolism , Opioid Peptides/metabolism , Receptors, Opioid, delta/metabolism , Receptors, Opioid, mu/metabolism , Substance-Related Disorders/metabolism , Animals , Brain/drug effects , Brain/physiopathology , Humans , Limbic System/drug effects , Limbic System/metabolism , Limbic System/physiopathology , Narcotics/pharmacology , Neural Pathways/drug effects , Neural Pathways/metabolism , Neural Pathways/physiopathology , Receptors, Opioid, delta/drug effects , Receptors, Opioid, mu/drug effects , Reward , Substance-Related Disorders/drug therapy , Substance-Related Disorders/physiopathologyABSTRACT
Drug addiction is a chronic relapsing disease in which drug administration becomes the primary stimulus that drives behavior regardless of the adverse consequence that may ensue. As drug use becomes more compulsive, motivation for natural rewards that normally drive behavior decreases. The discontinuation of drug use is associated with somatic signs of withdrawal, dysphoria, anxiety, and anhedonia. These consequences of drug use are thought to contribute to the maintenance of drug use and to the reinstatement of compulsive drug use that occurs during the early phase of abstinence. Even, however, after prolonged periods of abstinence, 80-90% of human addicts relapse to addiction, suggesting that repeated drug use produces enduring changes in brain circuits that subserve incentive motivation and stimulus-response (habit) learning. A major goal of addiction research is the identification of the neural mechanisms by which drugs of abuse produce these effects. This article will review data showing that the dynorphin/kappa-opioid receptor (KOPr) system serves an essential function in opposing alterations in behavior and brain neurochemistry that occur as a consequence of repeated drug use and that aberrant activity of this system may not only contribute to the dysregulation of behavior that characterizes addiction but to individual differences in vulnerability to the pharmacological actions of cocaine and alcohol. We will provide evidence that the repeated administration of cocaine and alcohol up-regulates the dynorphin/KOPr system and that pharmacological treatments that target this system may prove effective in the treatment of drug addiction.
Subject(s)
Dynorphins/metabolism , Receptors, Opioid, kappa/metabolism , Substance-Related Disorders/physiopathology , Alcoholism/physiopathology , Animals , Behavior, Addictive/physiopathology , Brain/metabolism , Cocaine/pharmacology , Cocaine/toxicity , Cocaine-Related Disorders/physiopathology , Ethanol/pharmacology , Ethanol/toxicity , Humans , Up-RegulationABSTRACT
Quantitative and conventional microdialysis were used to investigate the effects of constitutive deletion of the prodynorphin gene on basal dopamine (DA) dynamics in the nucleus accumbens (NAc) and the responsiveness of DA neurons to an acute cocaine challenge. Saline- and cocaine-evoked locomotor activity were also assessed. Quantitative microdialysis revealed that basal extracellular DA levels were decreased, while the DA extraction fraction, an indirect measure of DA uptake, was unchanged in dynorphin (DYN) knockout (KO) mice. The ability of cocaine to increase NAc DA levels was reduced in KO. Similarly, cocaine-evoked locomotor activity was decreased in KO. The selective kappa opioid receptor agonist U-69593 decreased NAc dialysate DA levels in wildtype mice and this effect was enhanced in KO. Administration of the selective kappa opioid receptor (KOPr) antagonist nor-binaltorphimine to KO mice attenuated the decrease in cocaine-induced DA levels. However, it was ineffective in altering the decreased locomotor response to cocaine. These studies demonstrate that constitutive deletion of prodynorphin is associated with a reduction of extracellular NAc DA levels and a decreased responsiveness to acute cocaine. Data regarding the effects of U-69593 and nor-binaltorphimine in KO suggest that the kappa opioid receptor is up-regulated as a consequence of prodynorphin gene deletion and that this adaptation underlies the decrease in basal DA dynamics and cocaine-evoked DA levels observed in DYN KO mice. These findings suggest that the phenotype of DYN KO mice is not solely due to loss of endogenous opioid peptide but also reflects developmental compensations that occur at the level of the opioid receptor.
Subject(s)
Cocaine/pharmacology , Dopamine Uptake Inhibitors/pharmacology , Dopamine/metabolism , Dynorphins/deficiency , Nucleus Accumbens/drug effects , Analgesics/pharmacology , Animals , Area Under Curve , Behavior, Animal , Benzeneacetamides/pharmacology , Chromatography, High Pressure Liquid/methods , Dose-Response Relationship, Drug , Drug Interactions , Electrochemistry/methods , Mice , Mice, Inbred C57BL , Mice, Knockout , Microdialysis/methods , Motor Activity/drug effects , Naltrexone/analogs & derivatives , Naltrexone/pharmacology , Narcotic Antagonists/pharmacology , Nucleus Accumbens/physiology , Pyrrolidines/pharmacologyABSTRACT
Conventional brain microdialysis was used to assess basal and cocaine-induced dopamine (DA) levels in the nucleus accumbens of wildtype (WT) C57BL/6J mice and mice with constitutive deletion of ether mu- or delta-opioid receptors (MOR or DOR knockout [KO], respectively). Locomotor activity was assessed in these same animals. Basal locomotor activity of DOR KO was elevated relative to MOR KO, but did not differ from that of WT mice. DOR mice, but not WT or MOR KO, exhibited a significant increase in activity in response to an injection of saline. The acute administration of cocaine produced a dose-related increase in locomotor activity in the three genotypes. The locomotor activating effects of a low dose (10 mg/kg) of cocaine were enhanced in DOR KO mice whereas the locomotor activating effects of both a low and higher (20 mg/kg) dose of cocaine were reduced in MOR KO animals. Microdialysis studies revealed no difference between genotypes in basal DA levels. Acute administration of cocaine, but not saline, increased DA levels in WT and KO animals. Paradoxically, however, the magnitude of this effect was smaller in DOR KO as compared with that in either WT or MOR KO. These data indicate that constitutive deletion of either MOR or DOR results in contrasting effects upon responsiveness to cocaine, which is consistent with the distinct phenotypes previously described for these mutants.
Subject(s)
Brain Chemistry/drug effects , Brain/drug effects , Cocaine-Related Disorders/metabolism , Cocaine/pharmacology , Receptors, Opioid, delta/physiology , Receptors, Opioid, mu/physiology , Animals , Brain/metabolism , Brain/physiopathology , Brain Chemistry/physiology , Cocaine-Related Disorders/genetics , Cocaine-Related Disorders/physiopathology , Dopamine/metabolism , Dose-Response Relationship, Drug , Extracellular Fluid/drug effects , Extracellular Fluid/metabolism , Genotype , Mice , Mice, Knockout , Microdialysis , Motor Activity/drug effects , Motor Activity/physiology , Nucleus Accumbens/drug effects , Nucleus Accumbens/metabolism , Opioid Peptides/metabolism , Receptors, Opioid, delta/deficiency , Receptors, Opioid, delta/genetics , Receptors, Opioid, mu/deficiency , Receptors, Opioid, mu/geneticsABSTRACT
Behavioral immobility during forced swim is usually considered a consequence of inescapable stress, and is used to screen antidepressant drugs. However, immobility in this test may also result from inhibition of neural functions because of brain hypothermia due to body cooling. To explore this possibility, we measured brain temperature dynamics during a 10-min forced swim in cold (25 degrees C) and warm (37 degrees C) water and correlated brain temperatures with behavioral changes. Cold water forced swim resulted in significant brain hypothermia (-6-7 degrees C) and immobility, while no immobility was observed during warm water forced swim, when brain temperature transiently increased (0.5 degrees C) then decreased below baseline in the post-swim period. These data suggest that immobility, which rapidly develops during forced swim in cold water, may result from dramatic inhibition of neural functions because of severe brain hypothermia.
Subject(s)
Behavior, Animal/physiology , Brain/physiopathology , Hypothermia/psychology , Motor Activity/physiology , Swimming/physiology , Swimming/psychology , Animals , Brain Chemistry/physiology , Calcium Channels/physiology , Cold Temperature , Dopamine/metabolism , Hypothermia/physiopathology , Male , Mice , Neostriatum/metabolism , Neostriatum/physiology , Sodium Channels/physiologyABSTRACT
Despite an abundance of studies on mechanisms of behavioral sensitization, considerable uncertainty exists as to whether alterations in dopamine neurotransmission underlie the exacerbated behavioral effects of cocaine observed during the early stages of abstinence. One of the factors contributing to the uncertainty and controversy may be the limitations in utilized measurement techniques (mostly conventional microdialysis). The techniques of quantitative microdialysis under transient conditions and rotating disk electrode voltammetry were used to characterize basal dopamine dynamics as well as time-related changes in extracellular dopamine concentrations and dopamine uptake that occur in response to an acute drug challenge in control animals and animals with previous history of cocaine. Basal extracellular dopamine concentrations were unaltered on abstinence day 3 from repeated cocaine administration (5 days, 20 mg/kg, i.p.). The extraction fraction of dopamine, an indirect measure of dopamine uptake, was significantly lower in cocaine-sensitized animals relative to controls. These two facts, taken together, suggest that basal dopamine release is depressed in cocaine-sensitized animals on abstinence day 3. At the same time, a cocaine challenge decreased the extraction fraction and increased the extracellular dopamine concentration in both experimental groups. The magnitude of the increase in extracellular dopamine concentration was greater in cocaine-sensitized animals, while the ability of cocaine to decrease the extraction fraction was unaltered, suggesting that the increase in extracellular dopamine concentration reflects an increase in drug-evoked dopamine release. Moreover, cocaine-pretreated rats demonstrated greater depolarization-induced dopamine release and the ability of dopamine D(2) receptor agonist, quinpirole, to inhibit release was decreased in these animals. These data demonstrate that a cocaine treatment regimen resulting in behavioral sensitization is associated with a reduction in basal dopamine release, an enhancement in both cocaine and K(+)-evoked dopamine release, and a subsensitivity of dopamine D(2) autoreceptors that regulate dopamine release in the nucleus accumbens.
Subject(s)
Cocaine/pharmacology , Dopamine Uptake Inhibitors/pharmacology , Dopamine/metabolism , Nucleus Accumbens/drug effects , Nucleus Accumbens/metabolism , Substance Withdrawal Syndrome/metabolism , Animals , Cocaine/metabolism , Dopamine Uptake Inhibitors/metabolism , Extracellular Space/metabolism , Illicit Drugs/pharmacology , Male , Microdialysis , Rats , Rats, Sprague-Dawley , Time FactorsABSTRACT
An involvement of the D(3) dopamine receptor in the regulation of extracellular dopamine has been suggested. However, the mechanisms mediating this effect are unclear. We have used the technique of no net flux microdialysis under transient conditions to examine the influence of the D(3) -preferring agonist (+)-PD128907 upon extracellular dopamine levels in the nucleus accumbens of the mouse. (+)-PD 128907 (0.1 mg/kg intraperitoneally) significantly decreased extracellular dopamine. This decrease was associated with a marked increase in the extraction fraction, which suggests an increase in dopamine clearance. The ability of D(3) -preferring compounds to modulate dopamine uptake was investigated in vitro using rotating disk electrode voltammetry. (+)-PD 128907 (10 nm) significantly increased the initial clearance rate of 3 microm dopamine in rat nucleus accumbens tissue suspensions. Kinetic analysis revealed no change in the apparent K (m) of uptake but it showed a 33% increase in V (max). In contrast, the D(3) antagonist GR 103691 (10 nm) significantly decreased dopamine uptake. Consistent with the low levels of D(3) receptors in the dorsal striatum, neither compound affected uptake in tissue suspensions from this brain region. These data indicate that D(3) receptor activation increases dopamine uptake in the nucleus accumbens and suggest that this receptor subtype can regulate extracellular dopamine by modulating the DA transporter activity.
Subject(s)
Dopamine/metabolism , Extracellular Space/metabolism , Nucleus Accumbens/metabolism , Receptors, Dopamine D2/metabolism , Animals , Benzopyrans/pharmacology , Biphenyl Compounds/pharmacology , Dopamine/analysis , Dopamine/pharmacokinetics , Dopamine Agonists/pharmacology , Dopamine Antagonists/pharmacology , Dopamine D2 Receptor Antagonists , Dose-Response Relationship, Drug , Electrochemistry , Extracellular Space/chemistry , In Vitro Techniques , Injections, Intraperitoneal , Ligands , Male , Mice , Microdialysis , Nucleus Accumbens/drug effects , Oxazines/pharmacology , Piperazines/pharmacology , Receptors, Dopamine D2/agonists , Receptors, Dopamine D3ABSTRACT
An involvement of the D3 dopamine receptor in the modulation of extracellular dopamine concentrations is suggested by pharmacological studies. However, recent studies using D3 receptor knock out mice indicated that several functions previously attributed to the D3 receptor are mediated by other receptor types. In the present study, we used the no-net flux microdialysis technique to characterize: (i) basal dopamine dynamics in the ventral striatum of D3 knock out and wild type mice and (ii) the effects of the putative D3-receptor selective agonist (+)-PD 128907. Neither the extracellular dopamine concentration nor the in vivo extraction fraction, an indirect measure of basal dopamine uptake, differed between D3 knock out and wild type mice. Moreover, no differences in potassium (60 mM) or cocaine (5 or 20 mg/kg i.p.) evoked dopamine concentrations were detected between the two genotypes. However, intra-striatal or systemic administration of doses of (+)-PD 128907 that failed to modify dopamine concentrations in knock out mice significantly decreased dialysate dopamine concentrations in the wild type. Comparison of the concentration-response curve for (+)-PD 128907 revealed IC(25) values of 61 and 1327 nM in wild type and knock out mice, respectively, after intra-striatal infusions. Similar differences were obtained after systemic administration of the D3 preferring agonist (IC(25) 0.05 and 0.44 mg/kg i.p. in wild type and knock out mice, respectively). We conclude that the activation of the D3 receptor decreases extracellular dopamine levels and that, at sufficiently low doses, the effects of (+)-PD 128907 on extracellular dopamine are selectively mediated by the D3 receptor.
Subject(s)
Benzopyrans/pharmacology , Dopamine Agonists/pharmacology , Dopamine/metabolism , Oxazines/pharmacology , Receptors, Dopamine D2/agonists , Animals , Brain/anatomy & histology , Cocaine/pharmacology , Dopamine Uptake Inhibitors/pharmacology , Kinetics , Male , Mice , Mice, Knockout , Microdialysis , Potassium/pharmacology , Receptors, Dopamine D2/genetics , Receptors, Dopamine D3ABSTRACT
The repeated, intermittent use of cocaine and other drugs of abuse produces profound and often long-lasting alterations in behavior and brain chemistry. It has been suggested that these consequences of drug use play a critical role in drug craving and relapse to addiction. This article reviews the effects of psychostimulant administration on dopaminergic and excitatory amino acid neurotransmission in brain regions comprising the brain's motive circuit and provides evidence that the activation of endogenous kappa-opioid receptor systems in these regions opposes the behavioral and neurochemical consequences of repeated drug use. The role of this opioid system in mediating alterations in mood and affect that occur during abstinence from repeated psychostimulant use are also discussed.
Subject(s)
Amphetamine/pharmacology , Cocaine-Related Disorders/physiopathology , Cocaine/pharmacology , Dopamine Uptake Inhibitors/pharmacology , Receptors, Opioid, kappa/drug effects , Receptors, Opioid, kappa/physiology , Adaptation, Psychological , Affect , Animals , Basal Ganglia/drug effects , Basal Ganglia/physiology , Brain Chemistry/drug effects , Humans , Ligands , Receptors, Dopamine/drug effects , Receptors, Dopamine/physiology , Thalamic Nuclei/drug effects , Thalamic Nuclei/physiologyABSTRACT
RATIONALE: Since cues that predict aversive outcomes can elicit both avoidance and 20 kHz ultrasonic vocalizations (USVs) in adult rats, 20 kHz USVs may also index the conditioned aversive effects of drugs. OBJECTIVE: We evaluated whether exposure to compartments associated with drugs with aversive effects would selectively increase 20 but not 50 kHz USVs in rats. METHOD: Rats were injected with naloxone (NAL) or lithium chloride (LiCl) and placed in one compartment or with saline (VEH) and placed in another compartment for three 50-min conditioning sessions. 20 kHz USVs, 50 kHz USVs, and time spent in each chamber were recorded during subsequent 15-min testing sessions during which rats had access to both compartments (expt 1) or were confined to the drug- or VEH-paired compartment (expt 2). RESULTS: In expt 1, animals conditioned either with NAL (0.3 and 3.0 mg/kg) or LiCl (10 and 30 mg/kg) emitted increased 20 kHz USVs in the drug-paired compartment, relative to VEH-conditioned controls. Conditioning with high doses of both drugs also increased conditioned place aversion and decreased emission of 50 kHz USVs. In expt 2, restriction of animals to the compartment paired with high doses of NAL and LiCl also increased emission of 20 kHz USVs and decreased 50 kHz USVs, relative to VEH-conditioned controls. CONCLUSIONS: In rats, cues associated with drugs with aversive effects increase 20 kHz USVs and decrease 50 kHz USVs, suggesting that USVs may provide a useful model for predicting the conditioned aversive effects of drugs.
Subject(s)
Antimanic Agents/pharmacology , Conditioning, Psychological/drug effects , Lithium Chloride/pharmacology , Naloxone/pharmacology , Narcotic Antagonists/pharmacology , Ultrasonics , Vocalization, Animal/drug effects , Analgesics, Opioid/pharmacology , Animals , Conditioning, Psychological/physiology , Cues , Male , Morphine/pharmacology , Rats , Rats, Sprague-Dawley , Vocalization, Animal/physiologyABSTRACT
Previous studies showed that prior administration of kappa-opioid agonists decreased the development of sensitization to some of the behavioral effects of cocaine. The present study sought to determine whether the development of sensitization to cocaine's reinforcing effects was also sensitive to antagonism by kappa-opioid agonists. During a pretreatment phase, the kappa-opioid agonist, U69593 (0.0 or 0.32 mg/kg) was administered prior to (1) 2 daily injections of cocaine (0.0 or 20.0 mg/kg), or (2) cocaine or saline administered via a yoking procedure. Cocaine pretreatment decreased the latency to acquisition of cocaine self-administration. However, prior administration of U69593 during the pretreatment phase failed to attenuate the development of this sensitized response to cocaine's reinforcing effect. In other groups, the effect of acute U69593 pretreatment on the maintenance of cocaine self-administration was examined during a 10 hr session. During training and testing, a stimulus was associated with each self-administered cocaine infusion for one group whereas responding of another group was reinforced by a cocaine infusion alone. On the test day, pretreatment with U69593 (0.32 mg/kg) decreased responding during each hour of the 10 hr session for the group that was reinforced with cocaine plus the cocaine-associated stimulus. U69593 failed to produce a long-lasting disruption of cocaine self-administration for rats that were trained and tested without the cocaine-associated stimulus. These data suggest that the acquisition and maintenance of cocaine self-administration are differentially sensitive to manipulations of kappa-opioid systems. Further, the disruption of cocaine self-administration by U69593 may be due to interactions with mechanisms that underlie facilitative effects of stimuli that have been associated with self-administered cocaine infusions.
Subject(s)
Behavior, Addictive/metabolism , Benzeneacetamides , Cocaine-Related Disorders/metabolism , Cocaine/administration & dosage , Pyrrolidines/administration & dosage , Receptors, Opioid, kappa/agonists , Animals , Behavior, Animal/drug effects , Conditioning, Psychological/drug effects , Disease Models, Animal , Drug Administration Schedule , Injections, Intravenous , Male , Photic Stimulation , Rats , Rats, Sprague-Dawley , Reaction Time/drug effects , Reinforcement, Psychology , Self AdministrationABSTRACT
The technique of microdialysis enables the monitoring of neurotransmitters and other molecules in the extracellular environment. This method has undergone several modifications and is now widely used for sampling and quantitating neurotransmitters, neuropeptides, and hormones in the brain and periphery. This unit describes the principles of conventional and quantitative microdialysis as well as strategies in designing a dialysis experiment. It establishes the groundwork for the basic techniques of preparation, conduct, and analysis of dialysis experiments in rodents and subhuman primates. Although the methods described are those used for monitoring CNS function, they can be easily applied with minor modification to other organ systems.
Subject(s)
Microdialysis , Animal Structures/chemistry , Animals , Columbidae , Diffusion , Equipment Design , Humans , Inorganic Chemicals/analysis , Mammals , Mathematics , Microdialysis/instrumentation , Microdialysis/methods , Oncorhynchus mykiss , Organ Specificity , Organic Chemicals/analysis , Research DesignABSTRACT
Microdialysis is an established sampling technique for the in vivo measurement of a variety of substances in both blood and tissue. These include neurotransmitters and neuropeptides, enzymes, and electrolytes, as well as various hormones and pharmaceutical agents. More recently, microdialysis has been used to infuse exogenous as well as endogenous substances into the brain and spinal cord. In microdialysis, a semipermeable dialysis membrane is introduced into the fluid or tissue compartment to be sampled and perfused with physiological fluid. As a result of passive diffusion, molecules migrate across the membrane along their concentration gradient. Molecules found in high concentrations within the tissue compartment migrate across the membrane into the dialysis tubing where they can be collected for subsequent quantification, whereas molecules found in high concentrations within the membrane diffuse outward into the surrounding tissue compartment. This unit describes construction of concentric and side-by-side microdialysis probes, which differ with respect to size and inflow/outflow tube compositions, as well as a modification of a horizontal probe. This unit also covers methods for surgically implanting microdialysis probes in rats and mice and for conducting microdialysis in vitro and in vivo in rodents. Finally, procedures are provided for carrying out quantitative microdialysis techniques.
Subject(s)
Microdialysis/methods , Rats/metabolism , Animals , Brain Chemistry , Catheterization , Equipment Design , Implants, Experimental , Mice , Microdialysis/instrumentation , Stereotaxic TechniquesABSTRACT
Coadministration of kappa-opioid receptor agonists (kappa-agonists) with cocaine prevents alterations in dialysate dopamine (DA) concentration in the nucleus accumbens (Acb) that occur during abstinence from repeated cocaine treatment. Quantitative microdialysis was used to determine the mechanism producing these effects. Rats were injected with cocaine (20 mg/kg, i.p.), or saline, and the selective kappa-agonist U-69593 (0.32 mg/kg, s.c.), or vehicle, once daily for 5 d. Extracellular DA concentration (DA(ext)) and extraction fraction (E(d)), an indirect measure of DA uptake, were determined 3 d later. Repeated cocaine treatment increased E(d), whereas repeated U-69593 treatment decreased E(d), relative to controls. Coadministration of both drugs yielded intermediate E(d) values not different from controls. In vitro DA uptake assays confirmed that repeated U-69593 treatment produces a dose-related, region-specific decrease in DA uptake and showed that acute U-69593 administration increases DA uptake in a nor-binaltorphimine reversible manner. Repeated U-69593 also led to a decrease in [(125)I]RTI-55 binding to the DA transporter (DAT), but did not decrease total DAT protein. These results demonstrate that kappa-opioid receptor activation modulates DA uptake in the Acb in a manner opposite to that of cocaine: repeated U-69593 administration decreases the basal rate of DA uptake, and acute U-69593 administration transiently increases DA uptake. kappa-agonist treatment also alters DAT function. The action of kappa-agonists on DA uptake or DAT binding, or both, may be the mechanism(s) mediating the previously reported "cocaine-antagonist" effect of kappa-opioid receptor agonists.
Subject(s)
Benzeneacetamides , Cocaine/antagonists & inhibitors , Dopamine/pharmacokinetics , Membrane Glycoproteins , Membrane Transport Proteins , Naltrexone/analogs & derivatives , Nerve Tissue Proteins , Nucleus Accumbens/drug effects , Nucleus Accumbens/metabolism , Receptors, Opioid, kappa/metabolism , Analysis of Variance , Animals , Autoradiography , Carrier Proteins/metabolism , Cocaine/administration & dosage , Corpus Striatum/chemistry , Corpus Striatum/metabolism , Dopamine/analysis , Dopamine Plasma Membrane Transport Proteins , Drug Administration Schedule , Drug Antagonism , Extracellular Space/chemistry , Extracellular Space/metabolism , Injections, Intraperitoneal , Injections, Subcutaneous , Iodine Radioisotopes , Linear Models , Male , Microdialysis , Naltrexone/administration & dosage , Nucleus Accumbens/chemistry , Pyrrolidines/administration & dosage , Rats , Rats, Sprague-Dawley , Receptors, Opioid, kappa/agonistsABSTRACT
In vivo microdialysis was used to characterize basal dopamine dynamics and cocaine-evoked dopamine levels in the medial prefrontal cortex of male Sprague-Dawley rats that had previously received once daily injections of cocaine (days 1-5; 20mg/kg, i.p.) in combination with the selective kappa-opioid receptor agonist U-69593 (days 3-5; 0.32mg/kg, s.c.) or its vehicle. The influence of these treatments on [3H]dopamine uptake in medial prefrontal cortex synaptosomes was also determined. Three days following the cessation of drug treatment, animals with prior history of cocaine administration exhibited enhanced psychomotor stimulation in response to a subsequent cocaine challenge. This effect was not apparent in animals that had previously received the cocaine treatment regimen in combination with the kappa-opioid receptor agonist U-69593. Cocaine challenge increased prefrontal dopamine levels in all pretreatment groups, but cocaine-pre-exposed animals had lower cocaine-evoked dopamine levels and higher basal in vivo extraction fraction, indicative of an increase in basal dopamine uptake relative to controls. Pretreatment with U-69593 prevented these effects of cocaine. Measurement of [3H]dopamine uptake in synaptosomes revealed a significant increase in uptake three days after the cessation of cocaine treatment. No increase in uptake was observed in animals that had received the cocaine treatment regimen in combination with U-69593. These results demonstrate that the early phase of abstinence from cocaine is associated with marked alterations in medial prefrontal cortex dopamine neurotransmission and that these neuroadaptations are prevented by the activation of kappa-opioid receptors. Furthermore, they raise the possibility that mesocortical dopamine neurons may be an important neural substrate upon which kappa-opioid agonists act to prevent the development of cocaine-induced behavioral sensitization.
Subject(s)
Benzeneacetamides , Cocaine-Related Disorders/metabolism , Cocaine/pharmacology , Dopamine/metabolism , Neural Pathways/drug effects , Neurons/drug effects , Prefrontal Cortex/drug effects , Receptors, Opioid, kappa/agonists , Substance Withdrawal Syndrome/metabolism , Synaptic Transmission/drug effects , Ventral Tegmental Area/drug effects , Analgesics/pharmacology , Animals , Cocaine-Related Disorders/pathology , Cocaine-Related Disorders/physiopathology , Drug Interactions/physiology , Extracellular Space/drug effects , Extracellular Space/metabolism , Male , Microdialysis , Motor Activity/drug effects , Motor Activity/physiology , Neural Pathways/cytology , Neural Pathways/metabolism , Neurons/cytology , Neurons/metabolism , Prefrontal Cortex/cytology , Prefrontal Cortex/metabolism , Pyrrolidines/pharmacology , Rats , Rats, Sprague-Dawley , Receptors, Opioid, kappa/metabolism , Substance Withdrawal Syndrome/pathology , Substance Withdrawal Syndrome/physiopathology , Synaptic Transmission/physiology , Synaptosomes/drug effects , Synaptosomes/metabolism , Ventral Tegmental Area/cytology , Ventral Tegmental Area/metabolismABSTRACT
Sensitization to cocaine has been attributed to alterations in excitatory amino acid and dopamine neurotransmission in the mesolimbic system. The present study sought to determine whether inhibition of NAALADase, an enzyme that cleaves glutamate from the endogenous neuropeptide, N-acetyl-aspartyl-glutamate (NAAG), attenuates sensitization to the psychomotor stimulant effects of cocaine. Rats received daily injections of cocaine (20.0 mg/kg/day; i.p.) or saline for 5 days. Fifteen minutes prior to these injections they received an i.p. injection of the NAALADase inhibitor, 2-PMPA (50.0-100 mg/kg), or vehicle. Locomotor activity and stereotypy produced by a challenge dose of cocaine (15.0 mg/kg) were assessed 3 days later. Acute cocaine administration increased locomotor activity in control animals. In animals with a prior history of cocaine administration, the behavioral response to cocaine was significantly enhanced. In animals that had received 2-PMPA in combination with cocaine, the enhancement of cocaine-induced locomotor activity was attenuated. No alteration in cocaine-evoked activity was observed in animals that had received once daily injections of 2-PMPA, alone. Acute administration of 2-PMPA also did not modify saline-induced locomotor activity or activity produced by an acute cocaine challenge. These data demonstrate that NAALADase inhibition attenuates the development of sensitization to the locomotor-activating effects of cocaine. Furthermore, this action cannot be attributed to an antagonism of the acute effects of cocaine.
Subject(s)
Carboxypeptidases/antagonists & inhibitors , Cocaine/pharmacology , Dopamine Uptake Inhibitors/pharmacology , Motor Activity/drug effects , Animals , Glutamate Carboxypeptidase II , Male , Motor Activity/physiology , Organophosphorus Compounds/pharmacology , Rats , Rats, Sprague-DawleyABSTRACT
OBJECTIVES: An unbiased place preference conditioning procedure was used to examine the influence of the non-opioid peptide, dynorphin A 2-17 (DYN 2-17), upon the conditioned and unconditioned effects of opiate withdrawal in the rat. METHODS: Rats were implanted SC with two pellets containing 75 mg morphine or placebo. Single-trial place conditioning sessions with saline and the opioid receptor antagonist naloxone (0.1-1.0 mg/kg; SC) commenced 4 days later. Ten minutes before SC injections, animals received an IV infusion of saline or DYN 2-17 (0.1-5.0 mg/kg). Additional groups of placebo- and morphine-pelleted animals were conditioned with saline and DYN 2-17. During each 30-min conditioning session, somatic signs of withdrawal were quantified. Tests of place conditioning were conducted in pelleted animals 24 h later. RESULTS: Naloxone produced wet-dog shakes, body weight loss, ptosis and diarrhea in morphine-pelleted animals. Morphine-pelleted animals also exhibited significant aversions for an environment previously associated with the administration of naloxone. These effects were not observed in placebo-pelleted animals. DYN 2-17 pretreatment resulted in a dose-related attenuation of somatic withdrawal signs. However, conditioned place aversions were still observed in morphine-pelleted animals that had received DYN 2-17 in combination with naloxone. Furthermore, the magnitude of this effect did not differ from control animals. CONCLUSIONS: These data demonstrate that the administration of DYN 2-17 attenuates the somatic, but not the conditioned aversive effects of antagonist-precipitated withdrawal from morphine in the rat. Differential effects of this peptide in modulating the conditioned and unconditioned effects of opiate withdrawal are suggested.