Kathleen M. Kantak

Evaluation of anti-cocaine antibodies and a cocaine vaccine in a rat self-administration model

Abstract Rationale: Previous pre-clinical studies with an anti-cocaine monoclonal antibody left open several issues critical to assessing the effectiveness of a vaccine for altering cocaine self-administration behavior. Objectives: The objectives of this study were to determine, first, whether changes in self-administration behavior would be systematically related to antibody level and, second, how the antibody affected the self-administration of different doses of cocaine. Methods: Two experiments were conducted using a second-order schedule of drug delivery in rats. The first was a passive-administration study using the anti-cocaine monoclonal antibody MO240 to examine the relationship between antibody level and cocaine self-administration behavior, and the second was an active-immunization study to examine the efficacy of the cocaine vaccine IPC-1010 for blocking various doses of self-administered cocaine. Results: The passive-administration experiment with control and 4-mg or 12-mg MO240 treatments showed that antagonism of the 1 mg/kg cocaine training dose was dependent on antibody level. In animals whose serum antibody levels were sustained above 0.05 mg/ml, there was a sufficient amount of antibody to reduce drug-seeking behavior and drug intake. In the active-immunization experiment, the cocaine vaccine IPC-1010 induced average serum antibody levels of 0.08 mg/ml and reduced the reacquisition of behavior by 1 mg/kg cocaine. Antagonism of cocaine self-administration after immunization was evident across a range of doses of cocaine and was only apparent in animals whose serum antibody levels exceeded 0.05 mg/ml. Furthermore, there was no evidence that the antagonism was surmountable within the dose range examined (up to 5.6 mg/kg). Conclusions: Antagonism of cocaine self-administration across a range of doses is feasible after immunization with a cocaine vaccine as long as antibody levels are of a sufficient concentration.

Context‐dependent prefrontal cortex regulation of cocaine self‐administration and reinstatement behaviors in rats

Evidence of stimulus attribute‐specificity within the prefrontal cortex (PFC) suggests that different prefrontal subregions may contribute to cocaine addiction in functionally distinct ways. Thus, the present study examined the effects of lidocaine‐induced inactivation of two distinct PFC subregions, the prelimbic (PL) or dorsal agranular insular (AId) cortices, on drug‐seeking and drug‐taking behaviors under cocaine maintenance and reinstatement testing conditions in rats trained to self‐administer 1 mg/kg cocaine under a second‐order schedule of drug delivery. Throughout maintenance and reinstatement phases, rats were exposed to conditioned light cues and contextual odor or sound cues. Results showed that PL inactivation during maintenance test sessions significantly reduced drug‐seeking and drug‐taking behaviors, and disrupted patterns of responding in rats exposed to light–sound, but not light–odor, cues. Moreover, lidocaine‐induced inactivation of the PL significantly attenuated drug‐seeking behavior during cue‐induced and cocaine prime‐induced reinstatement in rats exposed to light–sound cues only. In contrast, AId inactivation significantly attenuated cue‐induced reinstatement of drug‐seeking behavior in rats exposed to light–odor cues only. Drug‐seeking and drug‐taking behaviors in these rats were not disrupted during maintenance and cocaine prime‐induced reinstatement testing regardless of the type of contextual cues used. Together, these data suggest that PL and AId subregions play separate yet overlapping roles in regulating cocaine addiction in rats in ways that are dependent on the presence or absence of cocaine and on the types of contextual cues present in the cocaine self‐administration environment.

D-cycloserine Deters Reacquisition of Cocaine Self-Administration by Augmenting Extinction Learning

Augmentation of cue exposure (extinction) therapy with cognitive-enhancing pharmacotherapy may offer an effective strategy to combat cocaine relapse. To investigate this possibility at the preclinical level, rats and squirrel monkeys were trained to self-administer cocaine paired with a brief visual cue. Lever pressing was subsequently extinguished by withholding cocaine injections while maintaining response-contingent presentations of the cue. The glycine partial agonist D-cycloserine (DCS; 15 and 30 mg/kg in rats, 3 and 10 mg/kg in monkeys) was evaluated for its effects on the rate of extinction and subsequent reacquisition of cocaine self-administration. Compared with vehicle, pretreatment with 30 mg/kg DCS 0.5 h before extinction training reduced the number of responses and latency to reach the extinction criterion in rats, but neither dose of DCS altered these measures in monkeys. In both species, pretreatment with the higher dose of DCS before extinction training significantly attenuated reacquisition of cocaine self-administration compared with either extinction training in the absence of DCS or DCS in the absence of explicit extinction. Furthermore, treatment with 30 mg/kg DCS accompanied by brief handling (a stress induction) immediately after but not 6 h after extinction training attenuated reacquisition of cocaine self-administration in rats. No adverse effects of 10 mg/kg DCS were evident in quantitative observational studies in monkeys. The results suggest that DCS augmented consolidation of extinction learning to deter reacquisition of cocaine self-administration in rats and monkeys. The results suggest that DCS combined with exposure therapy may constitute a rational strategy for the clinical management of cocaine relapse.

Advancing the spontaneous hypertensive rat model of attention deficit/hyperactivity disorder.

To advance the spontaneous hypertensive rat (SHR) model of attention deficit/hyperactivity disorder (ADHD), experiments examined the SHR in tasks recognized to assess functioning of the prefrontal cortex or dorsal striatal. Tasks included odor-delayed win-shift (nonspatial working and reference memory), win-stay (habit learning), and attentional set-shifting (attention and behavioral flexibility). In Experiment 1, the SHR strain was compared with Wistar-Kyoto (WKY) and Wistar-Kyoto Hypertensive (WKHT) strains on the first 2 tasks. In Experiment 2, oral methylphenidate (1.5 mg/kg) and vehicle (water) were evaluated on all 3 tasks in SHR and WKY strains. Results demonstrated that the SHR made significantly more errors in the odor-delayed win-shift, win-stay, and attentional set-shifting tasks compared with the WKY. Similar performances in the WKY and WKHT indicated that deficits observed in the SHR were not related solely to hypertension. Treating the SHR with methylphenidate eliminated strain differences in all 3 tasks. These findings provide evidence that the SHR is a valid model for studying ADHD-associated neurocognitive deficits. Moreover, the current behavioral approach is appropriate to assess novel medications developed to target ADHD-associated neurocognitive deficits.

Hippocampal memory system function and the regulation of cocaine self-administration behavior in rats

There is considerable interest in elucidating neurocognitive mechanisms of cocaine addiction. This report focuses on the hippocampal memory system. Using food reward, two cognitive tasks were examined after lidocaine inactivation of the dorsal (dSUB) or ventral (vSUB) subiculum, the primary hippocampal output regions in rats. These tasks were conducted to first identify functionally relevant stereotaxic coordinates within the hippocampal memory system, in order to then examine its role in regulating drug-seeking and drug-taking behavior studied under a contextually discriminable FI 5-min(FR5:S) second-order schedule of cocaine and brief stimulus delivery. Inactivation of the dSUB and vSUB with 10microg lidocaine impaired hippocampal-dependent win-shift performance. Amygdalar-dependant conditioned cue preference, used as a test for behavioral specificity of lidocaine, was not affected following inactivation of either site. Inactivation of the dSUB with 100 microg lidocaine significantly reduced drug-seeking and drug-taking behavior studied during the cocaine self-administration maintenance phase. Following extinction, inactivation of neither the dSUB nor vSUB influenced reinstatement of drug-seeking behavior induced by drug-paired cues presented alone or with a cocaine priming injection. The impairments in win-shift performance are consistent with the spatial processing functions of the dSUB and vSUB, and the reduction in drug-taking behavior is consistent with the non-spatial temporal processing functions of the dSUB. The lack of an effect of dSUB and vSUB inactivation on reinstatement of drug-seeking behavior may relate to the fact that the contextual associations with cocaine were well-practiced at the time of cue reinstatement testing, and therefore, drug-seeking behavior was likely regulated by nonhippocampal-dependent mechanisms.

Vaccines against drugs of abuse: A viable treatment option?

Drug addiction is a chronically relapsing brain disorder. There is an urgent need for new treatment options for this disease because the relapse rate among drug abusers seeking treatment is quite high. During the past decade, many groups have explored the feasibility of using vaccines directed against drugs of abuse as a means of eliminating illicit drug use as well as drug overdose and neurotoxicity.Vaccines work by inducing drug-specific antibodies in the bloodstream that bind to the drug of abuse and prevent its entry into the brain. The majority of work in this area has been conducted with vaccines and antibodies directed against cocaine and nicotine. On the basis of preclinical work, vaccines for cocaine and nicotine are now in clinical trials because they can offer long-term protection with minimal treatment compliance. In addition, vaccines and antibodies for phencyclidine, methamphetamine and heroin abuse are currently under development. An underlying theme in this research is the need for high concentrations of circulating drug-specific antibodies to reduce drug-seeking and drug-taking behaviour when the drug is repeatedly available, especially in high doses.Although vaccines against drugs of abuse may become a viable treatment option, there are several drawbacks that need to be considered. These include: (i) a lack of protection against a structurally dissimilar drug that produces the same effects as the drug of choice; (ii) a lack of an effect on drug craving that predisposes an addict to relapse; and (iii) tremendous individual variability in antibody formation. Forced or coerced vaccination is not likely to work from a scientific perspective, and also carries serious legal and ethical concerns.All things considered, vaccination against a drug of abuse is likely to work best with individuals who are highly motivated to quit using drugs altogether and as part of a comprehensive treatment programme. As such, the medical treatment of drug abuse will not be radically different from treatment of other chronic diseases.

Cognitive task performance after lidocaine-induced inactivation of different sites within the basolateral amygdala and dorsal striatum.

To determine whether discrete components of amygdaloid and striatal memory systems could interact to guide behavior in a radial arm maze, conditioned cue preference (CCP) and win-stay accuracy were examined after lidocaine inactivation of either the rostral (rBLA) or caudal (cBLA) basolateral amygdala, the lateral (lDST) or medial (mDST) dorsal striatum, or a control site in rats. CCP expression was blocked only after rBLA or cBLA inactivation. lDST inactivation prevented attainment of criteria win-stay performance, whereas rBLA and mDST inactivation delayed it. Control site inactivation did not influence performance in either task. These findings suggest that the amygdala works independently of other memory systems to regulate learned responses in the CCP task, the rBLA may work cooperatively with the lDST to guide behavior in the win-stay task, and the mDST is less critical than the lDST for attaining criteria performance in the win-stay task.

Magnesium deficiency alters aggressive behavior and catecholamine function.

Magnesium is an abundant mineral in the brain and is important for monoamine neurotransmitter synthesis and receptor binding. It should, therefore, have behavior-altering effects. Three experiments were conducted to determine the influence of magnesium deficiencies on aggressive behavior and catecholamine function in mice. There were concentration- and time-dependent reductions in offensive aggressive behavior with magnesium deficiencies. Defensive behavior was affected in a manner opposite to that of offensive behavior. Upon administration of low doses of apomorphine and l-amphetamine, less dopamine- and norepinephrine-related behavior occurred with less magnesium in the diet. These reductions also showed a time dependency. These data demonstrate that magnesium has an influence on aggressive behavior in mice. Also, a magnesium deficiency is capable of altering the potency of catecholamine stimulating drugs.

Cognitive Enhancers for Facilitating Drug Cue Extinction: Insights from Animal Models

Given the success of cue exposure (extinction) therapy combined with a cognitive enhancer for reducing anxiety, it is anticipated that this approach will prove more efficacious than exposure therapy alone in preventing relapse in individuals with substance use disorders. Several factors may undermine the efficacy of exposure therapy for substance use disorders, but we suspect that neurocognitive impairments associated with chronic drug use are an important contributing factor. Numerous insights on these issues are gained from research using animal models of addiction. In this review, the relationship between brain sites whose learning, memory and executive functions are impaired by chronic drug use and brain sites that are important for effective drug cue extinction learning is explored first. This is followed by an overview of animal research showing improved treatment outcome for drug addiction (e.g. alcohol, amphetamine, cocaine, heroin) when explicit extinction training is conducted in combination with acute dosing of a cognitive-enhancing drug. The mechanism by which cognitive enhancers are thought to exert their benefits is by facilitating consolidation of drug cue extinction memory after activation of glutamatergic receptors. Based on the encouraging work in animals, factors that may be important for the treatment of drug addiction are considered.

Effects of serotonin receptor agonists and antagonists on offensive aggression in mice

The effects of serotonin receptor agonists 5-methoxytryptamine and quipazine, and antagonist mianserin on resident-intruder offensive aggression were investigated. Both agonists reduced aggression. The fact that 5-methoxytryptamine preferentially binds to 5-HT-1 receptors strongly suggest that the decreased aggression with S-methoxytryptamine was related to stimulation of an inhibitory 5-HT-1 receptor. It is also suggested that the reduction in aggression with quipazine was related to quipazines preferential binding to the 5-HT-1 receptor. The 5-HT-2 receptor antagonist mianserin reduced aggression suggesting that 5-HT-2 receptor blockade is inhibitory for aggression. Thus, two serotonin classes of receptors may be differentially involved in offensive aggression.