Pavlovian Conditioned Approach Research Articles

Amphetamine-induced incentive sensitization of sign-tracking behavior in adolescent and adult female rats.

Age-specific behavioral and neural characteristics may predispose adolescents to initiate and escalate use of alcohol and drugs. Adolescents may avidly seek novel experiences, including drugs of abuse, because of enhanced incentive motivation for drugs and natural rewards, perhaps especially when that incentive motivation is sensitized by prior drug exposure. Using a Pavlovian conditioned approach (PCA) procedure, sign-tracking (ST) and goal-tracking (GT) behavior was examined in amphetamine-sensitized and control adolescent and adult female Sprague-Dawley rats, with expression of elevated ST behavior used to index enhanced incentive motivation for reward-associated cues. Rats were first exposed to a sensitizing regimen of amphetamine injections (3.0 mg/kg/ml d-amphetamine per day) or given saline (0.9% wt/vol) once daily for 4 days. Expression of ST and GT was then examined over 8 days of PCA training consisting of 25 pairings of an 8-s presentation of an illuminated lever immediately followed by response-independent delivery of a banana-flavored food pellet. Results showed that adults clearly displayed more ST behavior than adolescents, reflected via both more contacts with, and shorter latencies to approach, the lever. Prior amphetamine sensitization increased ST (but not GT) behaviors regardless of age. Thus, when indexed via ST, incentive motivation was found to be greater in adults than adolescents, with a prior history of amphetamine exposure generally sensitizing incentive motivation for cues predicting a food reward regardless of age.

Antagonism at NMDA receptors, but not β-adrenergic receptors, disrupts the reconsolidation of pavlovian conditioned approach and instrumental transfer for ethanol-associated conditioned stimuli

Reconsolidation is the process by which memories require restabilisation following destabilisation at retrieval. Since even old, well-established memories become susceptible to disruption following reactivation, treatments based upon disrupting reconsolidation could provide a novel form of therapy for neuropsychiatric disorders based upon maladaptive memories, such as drug addiction. Pavlovian cues are potent precipitators of relapse to drug-seeking behaviour and influence instrumental drug seeking through at least three psychologically and neurobiologically distinct processes: conditioned reinforcement, conditioned approach (autoshaping) and conditioned motivation (pavlovian-instrumental transfer or PIT). We have previously demonstrated that the reconsolidation of memories underlying the conditioned reinforcing properties of drug cues depends upon NMDA receptor (NMDAR)- and β-adrenergic receptor (βAR)-mediated signalling. However, it is unknown whether the drug cue memory representations underlying conditioned approach and PIT depend upon the same mechanisms. Using orally self-administered ethanol as a reinforcer in two separate experiments, we investigated whether the reconsolidation of the memories underlying conditioned approach and PIT requires βAR- and NMDAR-dependent neurotransmission. For ethanol self-administering but non-dependent rats, the memories underlying conditioned approach and PIT for a pavlovian drug cue were disrupted by the administration of the NMDAR antagonist MK-801, but not the administration of the βAR antagonist propranolol, when given in conjunction with memory reactivation. As for natural reinforcers, NMDARs are required for the reconsolidation of all aspects of pavlovian drug memories, but βARs are only required for the memory representation underlying conditioned reinforcement. These results indicate the potential utility of treatments based upon disrupting cue-drug memory reconsolidation in preventing relapse.

Individual variation in the propensity to attribute incentive salience to an appetitive cue predicts the propensity to attribute motivational salience to an aversive cue

It has been proposed that animals that attribute high levels of incentive salience to reward-related cues may be especially vulnerable to addiction. Individual variation has also been observed in the motivational value attributed to aversive cues, which may confer vulnerability to anxiety disorders such as post-traumatic stress disorder (PTSD). There may be a core behavioral trait that contributes to individual variation in the motivational value assigned to predictive cues regardless of emotional valence. To test this hypothesis, we used a Pavlovian conditioned approach procedure to classify rats based on whether they learned to approach and interact with a cue predicting food reward (sign-trackers) or learned upon cue presentation to go to the location of impending food delivery (goal-trackers), and then examined Pavlovian fear conditioning in the same animals. It has recently been proposed that sign-trackers are more vulnerable to substance abuse because they attribute greater incentive motivational value to drug cues. Here we show that sign-trackers also have a tendency to be more fearful of discrete cues that predict footshock. In addition, we found that goal-trackers exhibited greater contextual fear when placed back into the original fear-conditioning context in the absence of temporally discrete cues. These results suggest that there may be a subset of individuals who tend to attribute high levels of motivational salience to predictive cues regardless of emotional valence, which may predispose them to a number of psychiatric comorbidities including PTSD and substance abuse. Other individuals use contexts to appropriately modify their reactions to such salient stimuli.

Novelty seeking, incentive salience and acquisition of cocaine self-administration in the rat

It has been suggested that incentive salience plays a major role in drug abuse and the development of addiction. Additionally, novelty seeking has been identified as a significant risk factor for drug abuse. However, how differences in the readiness to attribute incentive salience relate to novelty seeking and drug abuse vulnerability has not been explored. The present experiments examined how individual differences in incentive salience attribution relate to novelty seeking and acquisition of cocaine self-administration in a preclinical model. Rats were first assessed in an inescapable novelty task and a novelty place preference task (measures of novelty seeking), followed by a Pavlovian conditioned approach task for food (a measure of incentive salience attribution). Rats then were trained to self-administer cocaine (0.3 or 1.0 mg/kg/infusion) using an autoshaping procedure. The results demonstrate that animals that attributed incentive salience to a food-associated cue were higher novelty seekers and acquired cocaine self-administration more quickly at the lower dose. The results suggest that novelty-seeking behavior may be a mediator of incentive salience attribution and that incentive salience magnitude may be an indicator of drug reward.

A Cocaine Cue Acts as an Incentive Stimulus in Some but not Others: Implications for Addiction

In addicts drug cues attract attention, elicit approach, and motivate drug-seeking and drug-taking behavior, and addicts find it difficult to resist such cues. In preclinical studies we have found, however, that food cues acquire incentive motivational properties only in a subset of individuals. For example, a food cue becomes attractive, eliciting approach and engagement with it, and acts as an effective conditional reinforcer in some rats but not others. We asked, therefore, whether rats that have a propensity to attribute incentive salience to a food cue are the same ones that attribute incentive value to a drug (cocaine) cue. We first used a Pavlovian conditioned approach procedure to determine which individual rats attributed incentive salience to a food cue. A second cue was then associated with the IV self-administration of cocaine. Later, the ability of the cocaine cue to maintain self-administration behavior and to reinstate self-administration after extinction was assessed. We report that in individuals that had a propensity to attribute incentive salience to a food cue, a cocaine cue spurred motivation to take drugs (its removal greatly diminished self-administration) and reinstated robust drug-seeking after extinction. However, in those individuals that failed to attribute incentive salience to a food cue, the cocaine cue was relatively devoid of incentive motivational properties. We conclude that it is possible to determine, before any drug experience, which individuals will most likely have difficulty resisting drug cues, a trait that might confer susceptibility to addiction.

Effects of acute amphetamine exposure on two kinds of Pavlovian approach behavior

Two kinds of Pavlovian conditioned approach behavior are possible: approach of the CS (sign-tracking) and approach of the US (goal-tracking). We hypothesized that administration of AMP would increase sign-tracking and decrease goal-tracking. However, increasing doses of AMP (up to 2.0mg/kg) decreased measures of sign-tracking while simultaneously increasing measures of goal-tracking. Administration of AMP may shift responding from cues distant from the CS to cues closer to the CS.

Association learning-dependent increases in acetylcholine release in the rat auditory cortex during auditory classical conditioning

The cholinergic system has been implicated in sensory cortical plasticity, learning and memory. This experiment determined the relationship between the acquisition of a Pavlovian conditioned approach response (CR) to an auditory conditioned stimulus (CS) and the release of acetylcholine (ACh) in the primary auditory cortex in rats. Samples of ACh were collected via microdialysis during behavioral training in either an auditory classical conditioning task or in a non-associative control task. The conditioning group received daily pairings of a white noise CS with a sucrose pellet unconditioned stimulus (US), while the control group received an equal number of CS and US presentations, but with these stimuli being presented randomly. Training was conducted on three consecutive days, with microdialysis samples being collected on Days 1 and 3 in separate sub-groups. The level of ACh released in the auditory cortex during conditioning trials increased from the first to the third day of training in the conditioning group as rats acquired the CR, but did not change in the control group, which did not acquire a CR. These data provide direct evidence for the hypothesis that ACh release increases in the primary auditory cortex during natural memory formation, where cholinergic activation is known to contribute to the formation of specific associative representational plasticity in conjunction with specific memory formation.

The nucleus accumbens core and shell are critical for the expression, but not the consolidation, of Pavlovian conditioned approach

The nucleus accumbens (NAc) is important for the ability of motivationally significant stimuli to guide behavior. To further delineate its role in appetitive Pavlovian conditioning, we tested the hypothesis that the NAc contributes to memory consolidation and expression for a goal-tracking version of Pavlovian conditioned approach (PCA) in rats. We found that neither post-training reversible inactivation with the GABA receptor agonists muscimol and baclofen nor inhibition of protein synthesis with anisomycin (ANI) in either the core or shell regions of the NAc had an effect on approach to a reward port in response to a reward-predictive cue (conditioned stimulus, CS+). In contrast, pre-test reversible inactivation of both the core and shell decreased conditioned responding during the CS+. Unlike inactivation of the core, however, reversible inactivation of the shell also produced an increase in responding during the CS− and the inter-trial interval. This suggests that the NAc is not involved in the consolidation of goal-tracking PCA, but that once the memory is formed, the core is required for expression of the CS–unconditioned stimulus (US) association and the shell is required to inhibit conditioned approach behavior at times when the CS+ is not presented.

Neural dynamics underlying impaired autonomic and conditioned responses following amygdala and orbitofrontal lesions.

A neural model is presented that explains how outcome-specific learning modulates affect, decision-making, and Pavlovian conditioned approach responses. The model addresses how brain regions responsible for affective learning and habit learning interact and answers a central question: What are the relative contributions of the amygdala and orbitofrontal cortex to emotion and behavior? In the model, the amygdala calculates outcome value while the orbitofrontal cortex influences attention and conditioned responding by assigning value information to stimuli. Model simulations replicate autonomic, electrophysiological, and behavioral data associated with three tasks commonly used to assay these phenomena: Food consumption, Pavlovian conditioning, and visual discrimination. Interactions of the basal ganglia and amygdala with sensory and orbitofrontal cortices enable the model to replicate the complex pattern of spared and impaired behavioral and emotional capacities seen following lesions of the amygdala and orbitofrontal cortex.

Amphetamine Exposure Enhances Accumbal Responses to Reward-Predictive Stimuli in a Pavlovian Conditioned Approach Task

Acute and repeated exposure to psychostimulants such as amphetamine enhances the effects of pavlovian conditioned stimuli on conditioned behavior. It is hypothesized that amphetamine facilitates conditioned stimulus (CS) effects by selectively enhancing accumbal neuronal responses to stimuli. To test this hypothesis, rats were trained to discriminate between two pavlovian stimuli. One stimulus (i.e., CS+) was paired with sucrose delivery [i.e., unconditioned stimulus (US)], and the other stimulus (i.e., CS-) was paired with the absence of sucrose. Animals developed a conditioned approach response that occurred during the CS+ but not during the CS-. We tested the effect of different doses of amphetamine (0, 0.25, 0.5, or 1.0 mg/kg) on this conditioned approach behavior as well as on accumbal neuronal responses time locked to the CS+, the CS-, and the US. Acute amphetamine exposure increased conditioned approach behavior during the CS+, but not during the CS-. This change in behavior was associated with a selective increase in the magnitude of accumbal responses during the CS+. Repeated amphetamine administration followed by a drug-free period and reexposure did not affect the conditioned behavior, but increased accumbal responses to the CS+. These findings support the hypothesis that amphetamine exposure enhances behavioral responses to pavlovian conditioned stimuli by amplifying accumbal responses to those stimuli.

Rapid induction of Pavlovian approach to an ethanol-paired visual cue in mice

Although many studies have shown Pavlovian conditioned approach to cues paired with natural reinforcers, it has been quite difficult to induce such behavior with drug reinforcers. This experiment tested a novel Pavlovian procedure for inducing approach to a conditioned stimulus (CS) paired with ethanol. Mice (NZB/B1NJ, DBA/2J) received intraperitoneal injections of ethanol (2 g/kg) immediately before 10-min exposure to a rectangular chamber that contained a distinctive visual cue (star) at one end (Paired group, CS+ trials). On alternate days, saline injection preceded apparatus exposure with no distinctive cues (CS- trials). Unpaired control mice received ethanol in the home cage 60-75 min after each CS+ trial. NZB/B1NJ Paired group mice spent increasing amounts of time (>85% of the session) in proximity to the star, whereas Unpaired group mice did not. DBA/2J Paired group mice spent slightly more time on the star side than Unpaired group mice but did not show an acquisition curve. Postconditioning tests showed a strong preference for the star side in Paired groups from both strains after saline injection. However, only NZB/B1NJ mice showed a preference after ethanol. This study provides the first unambiguous demonstration of Pavlovian conditioned approach to an ethanol-paired visual stimulus in the absence of any contingency between the animal's behavior and drug exposure. This effect, which is remarkable both in terms of its magnitude and the rapidity with which it was produced (within 2-3 trials), may be related to the cue-associated craving that accompanies alcohol and drug addiction.

Post-training, but not post-reactivation, administration of amphetamine and anisomycin modulates Pavlovian conditioned approach

The psychostimulant, amphetamine (AMPH), and the protein synthesis inhibitor, anisomycin (ANI), have been shown to modulate the consolidation and reconsolidation of several types of learning. To determine whether Pavlovian conditioned approach (PCA) is modulated in a similar manner, we examined the effects of post-training and post-reactivation administration of both AMPH and ANI on memory for PCA. Male Long-Evans rats received PCA training sessions during which presentations of a CS+ were followed by sucrose delivery. AMPH (1 mg/kg, s.c.) injected immediately but not 6 h after the first training session enhanced PCA behavior. ANI (150 mg/kg, s.c.) injected immediately but not 3 h after the first training session impaired PCA behavior. This impairment was not due to the development of a conditioned taste aversion. To examine whether PCA can also be modulated by post-reactivation administration of AMPH and ANI, rats were given an injection of AMPH, ANI, or vehicle immediately after a memory reactivation session. Upon testing, the behavior of both the AMPH- and the ANI-treated rats was unaffected. This result remained consistent when the experiment was repeated with changes to various behavioral parameters (i.e., amount of training, length of memory reactivation). These findings indicate that AMPH and ANI act during the post-training but not the post-reactivation period to enhance and impair, respectively, the learning of PCA. This suggests that the consolidation of PCA can be modulated in a manner comparable to other types of learned associations, but once learned, the memory appears to be relatively robust and stable.

Individual differences in the propensity to approach signals vs goals promote different adaptations in the dopamine system of rats

The way an individual responds to cues associated with rewards may be a key determinant of vulnerability to compulsive behavioral disorders. We studied individual differences in Pavlovian conditioned approach behavior and examined the expression of neurobiological markers associated with the dopaminergic system, the same neural system implicated in incentive motivational processes. Pavlovian autoshaping procedures consisted of the brief presentation of an illuminated retractable lever (conditioned stimulus) followed by the response-independent delivery of a food pellet (unconditioned stimulus), which lead to a Pavlovian conditioned response. In situ hybridization was performed on brains obtained either following the first or last (fifth) day of training. Two phenotypes emerged. Sign-trackers (ST) exhibited behavior that seemed to be largely controlled by the cue that signaled impending reward delivery; whereas goal-trackers (GT) preferentially approached the location where the reward was delivered. Following a single training session, ST showed greater expression of dopamine D1 receptor mRNA relative to GT. After 5 days of training, GT exhibited greater expression levels of tyrosine hydroxylase, dopamine transporter, and dopamine D2 receptor mRNA relative to ST. These findings suggest that the development of approach behavior towards signals vs goal leads to distinct adaptations in the dopamine system. The sign-tracker vs goal-tracker phenotype may prove to be a valuable animal model to investigate individual differences in the way incentive salience is attributed to environmental stimuli, which may contribute to the development of addiction and other compulsive behavioral disorders.

Firing Patterns of Accumbal Neurons During a Pavlovian-Conditioned Approach Task

The nucleus accumbens (NAc) is necessary for the expression of Pavlovian-conditioned approach behavior but not for the expression of instrumental behavior conditioned in sessions that set a low response requirement. Although numerous studies have characterized firing patterns of NAc neurons in relation to instrumental behavior, very little is known about how NAc neurons encode information in Pavlovian tasks. In the present study, recordings of accumbal firing patterns were made during sessions in which rats performed a Pavlovian-conditioned approach task. Most of the recorded neurons (74/83, 89%) exhibited significant responses during the conditioned stimulus (CS) presentation and/or the reward exposure. The reward responses were prevalent, predominantly inhibitory, and comparable to reward responses observed in various types of behavioral paradigms, including instrumental tasks. The CS responses could be segregated into multiple subtypes on the basis of directionality, onset latency, and duration. Several characteristics of the CS firing patterns were unique relative to cue responses observed previously during alternative types of conditioning sessions. It is possible that the novel firing patterns correspond to the differential contributions of the accumbens to Pavlovian-conditioned approach behavior and instrumentally conditioned behavior. Regardless, the novel patterns of firing add to existing evidence that characterization of accumbal firing patterns in Pavlovian tasks may provide additional information about the neurophysiological mechanisms that mediate accumbal contributions to behavior.

Isolation rearing-induced facilitation of Pavlovian learning: Abolition by postsession intra-amygdala nafadotride

It has been shown previously in this laboratory that rats reared in social isolation acquire a Pavlovian-conditioned approach task much more rapidly than their respective controls. This study assessed the involvement specifically of the mesoamygdaloid dopamine pathway in this facilitated learning of isolates. Thus, animals were required to associate arbitrary stimuli with a pulsed light stimulus (unconditioned stimulus, US). The US, while without biological significance, was nevertheless capable of eliciting an intrinsic and sustained alerting response. Procedures ensured that the arbitrary stimuli (tone or clicker) did not elicit a response in the first instance, and were presented either paired (CS+) or unpaired (CS−) with the US. Isolates and socially reared controls received intra-amygdala infusions of the D3 dopamine receptor antagonist, l-nafadotride, or vehicle immediately following the end of each training session. The conditioned response increased over sessions in both groups of vehicle-infused rats during presentations of the CS+ stimulus, but not CS−, and isolates acquired this association more rapidly than controls. However, acquisition of this association was abolished by postsession intra-amygdala l-nafadotride. Responding to the US was largely unaffected by drug or rearing conditions. Hence, these data provide strong evidence for the specific involvement of the mesoamygdaloid dopamine projection in the facilitation of associative learning by isolation rearing.

Blockade of sensitisation-induced facilitation of appetitive conditioning by post-session intra-amygdala nafadotride

Prior d-amphetamine experience has been reported to enhance appetitive Pavlovian conditioning. The present study assessed the involvement of the mesoamygdaloid dopamine projection in this effect. Bilateral post-session intra-amygdala infusions of the D3 dopamine receptor antagonist, l-nafadotride, or vehicle were given during acquisition of a Pavlovian association in sensitised and unsensitised rats. During these sessions, subjects received presentations of a stimulus (CS +) paired with 10% sucrose availability. A second stimulus (CS −) was also presented but never paired with sucrose. Sensitised animals infused post-session with vehicle acquired a Pavlovian conditioned approach response during CS + presentations more rapidly than controls, as we have shown previously. However, post-session intra-amygdala l-nafadotride selectively retarded conditioned responding to the CS + in both groups of animals, abolishing the difference between sensitised and unsensitised rats. These results, therefore, extend the evidence for the involvement of the mesoamygdaloid dopamine projection in Pavlovian conditioning, and the facilitation of associative learning following sensitisation.

Differential Involvement of NMDA, AMPA/Kainate, and Dopamine Receptors in the Nucleus Accumbens Core in the Acquisition and Performance of Pavlovian Approach Behavior

Stimuli paired with primary rewards can acquire emotional valence and the ability to elicit automatic, Pavlovian approach responses that have been shown to be mediated by the nucleus accumbens. The present experiment investigated the effects of infusions of glutamatergic or dopaminergic receptor antagonists into the core of the nucleus accumbens on the acquisition and performance of Pavlovian discriminated approach to an appetitive conditioned stimulus. Rats were trained on an autoshaping task in which a conditioned stimulus (CS+; a lever) was inserted into the operant chamber for 10 sec, after which a food pellet was delivered. Presentation of another lever (CS-) was never followed by food. Subjects developed a conditioned response of approaching and contacting the CS+ selectively, although food delivery was not in any way contingent on the animals' response. A triple dissociation in the effects of AP-5, LY293558 [(3SR, 4aRS, 6RS, 8aRS)-6-[2-(iH-tetrazol-5-yl)ethyl]-1,2,3,4,4a,5,6,7,8,8a-decahydroiso-quinoline-3-carboxylic acid], and alpha-flupenthixol infused into the nucleus accumbens core on the acquisition and performance of this conditioned response was observed. The AMPA/kainate receptor antagonist LY293558 disrupted discriminated approach performance but not acquisition, as evidenced by increased approaches to the CS-. In contrast, the NMDA receptor antagonist AP-5 impaired only the acquisition, but not performance, of autoshaping whereas the dopamine D1/D2 receptor antagonist alpha-flupenthixol decreased approaches to the CS+ during both acquisition and performance. The data are discussed with reference to dissociable interactions of these receptor types with limbic cortical and dopaminergic afferents to the nucleus accumbens core during the acquisition and expression of Pavlovian conditioned approach.

Enhanced dopamine efflux in the amygdala by a predictive, but not a non-predictive, stimulus: facilitation by prior repeated d-amphetamine

Extracellular levels of dopamine within the amygdala were monitored using in vivo microdialysis during performance of an appetitive Pavlovian conditioning task in sensitized rats and unsensitized controls. Animals received exposure either to d-amphetamine or to vehicle for seven consecutive days (2 mg/kg/day, i.p.) in the home cage. Training began following a further seven injection-free days. Animals were exposed to two session types: during conditioning sessions, a stimulus (tone or light) immediately preceded sucrose pellet delivery. During control sessions, the alternative stimulus was also presented, but not in temporal proximity to an otherwise identical schedule of pellet delivery. There was a total of three alternating presentations of each session type during training. Sensitization enhanced Pavlovian conditioned approach behaviour to the stimulus predictive of imminent pellet delivery, and was without effect upon approach behaviours either to the food pellets themselves or to the control stimulus. Extracellular levels of dopamine within the amygdala were assessed during the fourth conditioning and control sessions. Mesoamygdaloid dopamine efflux increased significantly during the conditioning test session, but not during the control session, and this dopaminergic response was more marked in rats with prior repeated d-amphetamine experience. Hence, these results add to evidence suggesting a role for amygdaloid dopamine in appetitive Pavlovian conditioning, and in the facilitation of associative learning following prior experience of d-amphetamine.

Sexual Pavlovian conditioned approach behavior in male Japanese quail (Coturnix coturnix japonica).

A localized visual stimulus presented immediately prior to access to a female conspecific stimulated approach behavior in male Japanese quail after several conditioning trials. Development of this conditioned approach behavior was observed with two different types of signal lights, 10-s and 30-s signal durations, large and small experimental chambers, and with male birds housed continually in the conditioning chambers or only placed in the chamber for brief daily sessions. Conditioning also resulted in shorter latencies to initiate copulation in males given access to a female following the signal light, in comparison with males that received access to a female unannounced by the light. Although some aspects of the conditioning experience were retained over a 6-week interval, some loss of the behavior was also observed. The conditioned behavior also decreased with repeated extinction trials, during which the signal light was presented in the absence of access to a female conspecific. The results are discussed in relation to other aspects of the social behavior of Japanese quail and other studies of sexual conditioning.

Sexual Pavlovian conditioned approach behavior in male Japanese quail (Coturnix coturnix japonica).

Sexual Pavlovian conditioned approach behavior in male Japanese quail (Coturnix coturnix japonica).