Reduction of latent inhibition by D-amphetamine in a conditioned suppression paradigm in humans

The sensitivity of latent inhibition (LI) to amphetamine has been tested in humans with a paradigm close to the conditioned emotional response suppression currently used in experimental animals. The conditioned stimulus (CS) was a tone, the unconditioned stimulus (US) a strong white noise, and the response a transient delay in a regular sequence of hand movements in the resolution of the Tower of Toronto puzzle. The aim of this study was to verify whether the previously reported, disruptive effect of CS preexposure on conditioning really represents LI, by examining its sensitivity to amphetamine. Three groups of healthy volunteers received placebo, 5 or 10 mg of dexamphetamine sulphate, respectively, in a double-blind experimental design. The preexposure, conditioning and test phases were carried out under either amphetamine or placebo. The non preexposed groups treated with amphetamine were not different from the non preexposed placebo group, indicating that amphetamine did not affect conditioning. Among the preexposed groups, those receiving 10 mg of amphetamine showed normal rates of conditioning, whereas those treated with either 5 mg of amphetamine or placebo showed LI. Similar results have been reported in experimental animals. This sensitivity to amphetamine suggests that the present paradigm may be used to study LI in humans.     

Associative learning and latent inhibition in a conditioned suppression paradigm in humans

A paradigm based on conditioned suppression of ongoing motor activity, sensitive to latent inhibition (LI), was developed and tested in healthy volunteers. Subjects were trained to move disks from one peg to another with a high degree of regularity in the Tower of Toronto puzzle, a well-known cognitive skill learning task. Once this was achieved, they were submitted to a Pavlovian conditioning procedure. The conditioned stimulus (CS) was a pure tone and the unconditioned stimulus (US) a loud white noise. The resulting response suppression was assessed by a transient increase in latency of the hand movements. In control subjects, there was non-contingent CS and US presentation. The results evidenced conditioning after a single CS-US pairing. Following five preexposures to the to-be-conditioned CS, however, conditioning was abolished, seemingly expressing LI. Because a weak unconditioned response to the tone was observed after its first two presentations, an additional experiment was performed with two preexposures to the to-be-conditioned CS. With such procedure, conditioning was obtained, supporting the existence of LI in the preceding experiment. These results indicate that the present paradigm may be useful for the study of LI in human subjects, having the advantage of being similar to the experimental conditions used in the majority of LI studies in experimental animals.     

The basolateral complex of the amygdala is necessary for acquisition but not expression of CS motivational value in appetitive Pavlovian second-order conditioning

The basolateral complex of the amygdala (ABL) is involved in processing information about stimulus motivational value. However, it is not clear whether the ABL is critical for acquisition, maintenance, or expression of this information. Our previous work has shown that ABL lesions made prior to training, block acquisition of an appetitive Pavlovian second-order conditioning task, in which performance is thought to depend on the acquisition of motivational (conditioned reinforcement) value by the first-order conditioned stimulus (CS). The present experiments examined the effects of ABL lesions made after first-order conditioning, when the CS acquires motivational value, but before second-order conditioning, the test for acquired value of that CS. Rats received pairings of a visual CS with a food reinforcer. They then received bilateral sham or excitotoxic lesions of the ABL. After recovery, they received pairings of a second-order auditory CS with the previously conditioned visual CS. In two experiments, both sham and lesioned rats acquired normal second-order conditioned behaviours. Some of the same rats were then given another round of second-order conditioning with novel CSs. In this case, when first-order training occurred after surgery, some second-order conditioned behaviours were impaired in lesioned rats. Tests of the associative underpinnings of second-order conditioned behaviours showed that those behaviours impaired by ABL lesions were based on stimulus-response associations. The results show that although the ABL is critical for second-order conditioning, this role is limited to acquisition of information about the motivational value of the first-order CS, and it is not critical for maintenance of this information or for its use in forming second-order associations.     

Post-training excitotoxic lesions of the dorsal hippocampus attenuate forward trace,backward trace,and delay fear conditioning in a temporally specific manner

The present study sought to determine whether post-training excitotoxic lesions of the dorsal hippocampus would disrupt retention of fear conditioned using a trace procedure. Rats were trained using one of six procedures. Forward trace conditioning consisted of 10 trials in which a 16-s tone conditional stimulus (CS) was followed by a 28-s stimulus-free trace interval and then a mild footshock unconditional stimulus (US). We used two forms of delay conditioning where the tone and footshock co-terminated. Short delay used a 16-s tone and long delay used a 46-s tone. Backward trace conditioning was the same as forward trace, except that the order of the CS and US was reversed. CS-only and US-only were similar to forward trace except that the footshock or tone, respectively, was eliminated. One day later, animals received either an N-methyl-D-aspartate (NMDA)-induced lesion of the dorsal hippocampus or sham surgery. One week later, the rats were tested for freezing to the tone in a novel context. The next day, they were tested for freezing to the original training context. Hippocampal lesioned trace conditioned rats showed significantly less freezing during the tone compared with their sham lesioned controls. The lesion did not affect freezing during the tone in delay conditioning, nor in the other training conditions. During the 1-min period after tone offset, there was a trend in all hippocampal lesioned animals toward a deficit in freezing, compared with their corresponding sham lesioned controls, although only short delay, forward and backward trace groups showed a significant deficit. Hippocampal lesions also attenuated contextual conditioning. Thus, the hippocampus is critical for the consolidation and/or expression of a trace fear conditioned stimulus.     

Differential participation of the NBM in the acquisition and retrieval of conditioned taste aversion and Morris water maze

Deficits in both learning and memory after lesions of the cholinergic basal forebrain, in particular the nucleus basalis magnocellularis (NBM), have been widely reported. However, the participation of the cholinergic system in either acquisition or retrieval of memory process is still unclear. In this study, we tested the possibility that excitotoxic lesions of the NBM affect either acquisition or retrieval of two tasks. In the first experiment, animals were trained for two conditioned taste aversion tasks using different flavors, saccharine and saline. The acquisition of the first task was before NBM lesions (to test retrieval) and the acquisition of the second task was after the lesions (to test acquisition). Accordingly, in the first part of the second experiment, animals were trained in the Morris water maze (MWM), lesioned and finally tested. In the final part of this experiment, another set of animals was lesioned, then trained in the MWM and finally tested. All animals were able to retrieve conditioned taste aversion (CTA) and MWM when learned before NBM lesions; however, lesions disrupted the acquisition of CTA and MWM. The results suggest that the NBM and cholinergic system may play an important role in acquisition but not during retrieval of aversive memories.     

Abnormally persistent latent inhibition induced by lesions to the nucleus accumbens core, basolateral amygdala and orbitofrontal cortex is reversed by clozapine but not by haloperidol

Latent inhibition (LI) is the proactive interference of inconsequential preexposure to a stimulus with its ability to signal significant events, and disrupted LI is considered to model positive symptoms of schizophrenia. We have recently shown that lesions of the nucleus accumbens core (NACc), basolateral amygdala (BLA) and orbitofrontal cortex (OFC) produce abnormally persistent LI, and suggested that this phenomenon may model negative symptoms. Here we tested whether NACc, BLA and OFC lesion-induced persistent LI would be reversed by the atypical antipsychotic drug (APD) clozapine but not by the typical APD haloperidol. Because clozapine's action is likely reflecting its 5HT2A receptor antagonism, we also tested whether NACc lesion-induced persistent LI would be reversed by the selective 5HT2A antagonist M100907. LI was measured in a conditioned emotional response procedure by comparing suppression of drinking in response to a tone in rats receiving 0 (non-preexposed) or 40 tone presentations (preexposed) followed by five tone-shock pairings. Under these conditions, control rats did not show LI but all lesioned rats persisted in exhibiting LI, and this was reversed by clozapine but not by haloperidol. In addition, M100907 reversed NACc lesion-induced persistent LI. These two novel phenomena, abnormally persistent LI and its selective reversal by an atypical APD, suggest a novel index of schizophrenia relevant behavioral abnormality and of atypical antipsychotic activity in the LI model. The identification of brain regions whose damage leads to persistent LI in the rat may provide valuable cues on dysfunctional brain circuits involved in negative symptoms and in the action of atypical APDs.     

Hippocampectomy and feature-positive discrimination

The effects of hippocampal lesions on feature-positive discrimination were investigated using the nictitating membrane response preparation. During training, animals received a simultaneous reinforced compound as the conditioned stimulus (CS+) and a non-reinforced element as the CS-. The compound consisted of a tone and a light, with the tone being more salient than the light. The light and tone served as the CS- in Expts. 1 and 2, respectively. There were no significant differences between hippocampectomized animals and the controls (cortical and sham) when the CS- was the light; however, when the more salient tone stimulus was the CS-, hippocampectomized animals exhibited high levels of responding to both the CS+ and CS- and failed to acquire the discrimination. The results are discussed in terms of attentional and response inhibition theories of hippocampal function.     

Learned irrelevance is disrupted in first-episode but not chronic schizophrenia patients

Learned irrelevance (LIrr) is a pre-exposure effect in which uncorrelated presentations of a conditioned stimulus (CS) and an unconditioned stimulus (US) retard subsequent CS-US association. LIrr is closely related to the phenomenon of latent inhibition (LI). LI refers to the retarding effects of inconsequential stimulus pre-exposure on subsequent conditioning to that stimulus, and is considered to reflect the organism's capacity to ignore irrelevant stimuli. LI is disrupted in schizophrenia patients, due to faster learning of the association between the preexposed CS and the US. A new within-subject target-recognition LIrr procedure was applied. The target was either cued by a priming signal or appeared at random, and priming signals were novel or preexposed cues. Schizophrenia patients were compared to age- and sex-matched control subjects. Normal subjects (n = 24) have shown robust LIrr, namely, faster cue-target associations of novel compared to preexposed cues. Schizophrenia patients at the early stages of their first episode (n = 7) showed LIrr disruption, namely, cue-target associations to preexposed cues were as fast as for novel cues. Chronic patients during an acute phase (n = 18) did not show LIrr as they failed to learn the cue-target association. In addition to the LIrr paradigm the same subjects were tested in a covert-orientation task. No differences were observed between the groups on this task. The possible advantages of the new LIrr paradigm are discussed.     

The latent inhibition model of schizophrenic attention disorder. Haloperidol and sulpiride enhance rats' ability to ignore irrelevant stimuli

Latent inhibition (LI) is a behavioral paradigm in which prior exposure to a stimulus not followed by reinforcement retards subsequent conditioning to that stimulus when it is paired with reinforcement. The development of LI reflects a process of learning to ignore, or tune out, irrelevant stimuli. Two experiments investigated the effects of haloperidol (0.02, 0.1, and 0.5 mg/kg) and sulpiride (100 mg/kg) administration on LI. The investigation was carried out using a conditioned emotional response (CER) procedure consisting of three stages: (1) preexposure, in which the to-be-conditioned stimulus, tone, was repeatedly presented without reinforcement; (2) conditioning, in which the preexposed stimulus was paired with shock; and (3) test, where LI was indexed by animals' suppression of licking during tone presentation. The three stages were conducted 24 hr apart. In the preexposure stage, only ten nonreinforced stimulus preexposures were given, a procedure known to be insufficient to yield LI in normal animals. In both experiments, LI was indeed absent in the placebo animals. In marked contrast, animals treated with haloperidol (experiment 1) as well as with sulpiride (experiment 2) exhibited LI. These results demonstrate that both typical and atypical neuroleptics enhance animals' capacity to ignore irrelevant stimuli. The implications of this finding for an animal model of schizophrenia and for a novel screening test for antipsychotic drugs are discussed.     

Reversible inactivation of the entorhinal cortex disrupts the establishment and expression of latent inhibition of cued fear conditioning in C57BL/6 mice

For latent inhibition, preexposure to a conditioned stimulus (CS) prior to training with an unconditioned stimulus (US) results in decreased conditioned responses (CRs) to the CS at the time of testing. The mechanism by which decreased CRs occurs, however, is unknown; CS preexposure may interfere with subsequent conditioning, or modulate the expression of CRs. Previous research has suggested that the entorhinal cortex (EC) is necessary for latent inhibition of a variety of tasks. However, no studies have specifically compared the role of the EC in acquisition vs. expression of latent inhibition. The present study used reversible inactivation of the EC to address this issue. The GABA agonist muscimol (0.5 microg/side) was directly infused into the EC of mice prior to CS preexposure, training, or testing. Our results indicate that muscimol inactivation of the EC before CS preexposure disrupts latent inhibition of cued fear conditioning. Importantly, this same dose of muscimol did not disrupt cued fear conditioning, nor did it affect latent inhibition when infused into the subiculum. Furthermore, inactivation of the EC at testing disrupted the expression of latent inhibition of cued fear conditioning; that is, CS preexposed mice that received entorhinal cortical muscimol infusion at testing showed CRs compared to saline-infused CS preexposed mice. These findings suggest that repeated preexposure to the CS during latent inhibition may alter entorhinal cortical activity thereby allowing the EC to exert inhibitory control over the expression of CRs during testing of CS preexposed mice.     

Behavioural and cardiovascular responses during latent inhibition of conditioned fear: measurement by telemetry and conditioned freezing

This study assessed freezing behaviour and cardiovascular responses during the expression of latent inhibition of conditioned fear. Animals that were either repeatedly preexposed (PE) to a tone conditioned stimulus (CS) or naive to the tone (non-preexposed; NPE) subsequently experienced three presentations of the tone paired with footshock. Animals were tested 24 h later in the context of the footshock chamber, and on the following day, in the presence of the tone CS. Changes in heart rate and blood pressure were recorded by radio-telemetry. The PE rats spent more time freezing to the conditioned contextual cues and exhibited higher blood pressures during the last half of the context test session than did the NPE animals. During the tone test, the PE rats exhibited less conditioned freezing to the tone CS compared with the NPE animals, i.e. expression of the latent inhibition. This behavioural effect was associated with a significant increase in heart rate, but not blood pressure, in the PE but not the NPE animals. Our results suggest that the increased blood pressures of the PE rats during the context test directly reflect their greater fear of the conditioning context. In contrast, the increased heart rate response but decreased freezing shown by PE rats in response to the tone CS may be due to the fact that lower stress levels (e.g. PE condition) elicit sympathetically-mediated increases in heart rate, whereas higher stress levels (e.g. NPE condition) activate both sympathetic and parasympathetic systems, thus eliminating any CS-induced increase in heart rate in the NPE rats.     

Hippocampectomy disrupts the topography of the rat eyeblink response during acquisition and extinction of classical conditioning.

The effect of hippocampal lesions on acquisition and extinction of eyeblink conditioning in rats was examined with the procedure described by Schmajuk and Christiansen (Physiol. Behav., 48 (1990) 755-758). Male Long-Evans rats received either sham, cortical control or hippocampal lesions. Animals were trained in a delay conditioning paradigm with a 500-ms, 98-dB, white noise conditioned stimulus; a 150-ms, 5-psi air puff unconditioned stimulus; a 350-ms interstimulus interval; and a 60-s average intertrial interval. Each animal received 50 trials daily until it reached a criterion of 80% conditioned responses (CR) on a given day. After acquisition criterion was reached, extinction began the next day and continued until each animal achieved a criterion of one CR in a block of 10 consecutive trials. Although hippocampal lesions affected neither acquisition nor extinction rates, hippocampal lesioned animals showed significantly shorter CR onset latency during acquisition and extinction, and larger CR peak amplitude during acquisition. The results are discussed in terms of hippocampal modulation of cerebellar learning.     

Enhanced visual latent inhibition in obsessive-compulsive disorder.

BACKGROUND: Latent inhibition (LI) is the slowed acquisition of a learned response to a conditioned stimulus (CS), that occurs if that CS has previously been experienced in a noncontingent setting. This retarded acquisition is thought to occur because, due to the previous noncontingent experience, an individual must "unlearn the irrelevance" of the CS, before learning its new association to the unconditioned stimulus. A previous report using an auditory paradigm did not detect abnormal LI in obsessive-compulsive disorder (OCD) subjects; this auditory LI task included a difficult acquisition phase, which made it relatively insensitive to detecting abnormally elevated LI (i.e., slowed "unlearning"). METHODS: We assessed LI using a highly sensitive computerized visual LI paradigm in 63 carefully screened control subjects and in 48 patients with OCD. RESULTS: Compared to controls, OCD subjects exhibited significantly more LI; if "preexposed" to a to-be-CS, OCD subjects required significantly more trials to learn a new association to that CS, compared to control subjects. This pattern was particularly evident in unmedicated OCD subjects. CONCLUSIONS: The inflated impact of preexposure on LI response acquisition in OCD subjects may be a quantitative measure of their tendency to remain "stuck in set" in this cognitive task.     

Effects of microinjections of apomorphine and haloperidol into the inferior colliculus on the latent inhibition of the conditioned emotional response

Electrical or chemical stimulation of the inferior colliculus (IC) induces fear-like behaviors. More recently, consistent evidence has shown that electrical stimulation of the central nucleus of the IC supports Pavlovian conditioning and latent inhibition (LI). LI is characterized by retardation in conditioning and also by an impaired ability to ignore irrelevant stimuli, after a non-reinforced pre-exposure to the conditioned stimulus. LI has been proposed as a behavioral model of cognitive abnormalities seen in schizophrenia. The aim of the present study was to determine whether dopaminergic mechanisms in the IC are involved in LI of the conditioned emotional response (CER). To induce LI, a group of rats was pre-exposed (PE) to six tones in two sessions, while rats that were not pre-exposed (NPE) had two sessions without tone presentations. The conditioning consisted of two tone presentations to the animal, followed immediately by a foot shock. PE and NPE rats received IC microinjections of physiological saline, the dopaminergic agonist apomorphine (9.0 μg/0.5 μL/side), or the dopaminergic antagonist haloperidol (0.5 μg/0.5 μL/side) before both pre-exposure and conditioning. During the test, the PE rats that received saline or haloperidol had a lower suppression of the licking response compared to NPE rats that received vehicle or haloperidol, indicating that latent inhibition was induced. There was no significant difference in the suppression ratio in rats that received apomorphine injections into the IC, indicating reduced latent inhibition. These results suggest that dopamine-mediated mechanisms of the IC are involved in the development of LI.     

Effects of lesions of the dorsal noradrenergic bundle on conditioned suppression to a CS and to a contextual background stimulus

The aim of the experiment was to determine whether the dorsal noradrenergic bundle (DB) plays a role in conditioning to context. Rats received either bilateral lesions of the DB by local injection of 6-hydroxydopamine, vehicle injections only, or sham operations. All animals were then trained to barpress for food on a variable interval (VI) schedule. Two 5-min intrusion periods were superimposed on the VI baseline during each session. An 'envelope' stimulus (flashing light) was on throughout each intrusion period. In addition, embedded in the two intrusion periods of each session, there occurred 8 presentations of a 'punctate' conditioned stimulus (CS) (a 15-s clicker), and 8 presentations of a 0.5-s footshock. Within each surgical condition rats were randomly allocated to one of three conditioning groups, receiving 100%, 50% or 0% temporal association between CS and shock. Conditioning to the punctate CS and to the context provided by the envelope stimulus was assessed by the degree of suppression of the barpress response relative to the VI baseline. Responding was most suppressed in the punctate CS in the 100 and 50% conditions, and most suppressed in the envelope stimulus in the 0% condition. DB lesions released response suppression to the punctate CS, had no effect on suppression to the envelope stimulus, and reduced sensitivity to CS-shock probability as measured by response suppression during the punctate CS. These results confirm previous reports that DB lesions alleviate response suppression to shock-associated cues, identify some of the parameters that affect this phenomenon, but fail to support a role for the DB in contextual conditioning.     

Auditory cortex lesions prevent the extinction of Pavlovian differential heart rate conditioning to tonal stimuli in rabbits

The present study examined the role of the auditory cortex in the extinction of differentially conditioned heart rate (HR) responses in rabbits. Lesions were placed bilaterally in wither the auditory cortex or the visual cortex. Three days after recovery from surgery, the auditory cortex lesion group and the visual cortex lesion control group were habituated to the tonal conditioned stimuli (CSs), and then given 2 days of Pavlovian differential conditioning (60 trials per day) in which one tone (CS+) was always paired with the unconditioned stimulus and another tone (CS−) was never paired with the uncondition stimulus. Animals that had demonstrated reliable differential conditioning (CS+ response at least 5 beats greater than the CS− response) were placed on an extinction schedule for 7 days. The extinction schedule was identical to the differential conditioning schedule with the exception that shock never followed the CS+. The results of the study indicate that auditory cortex lesions prevent the extinction of differential bradycardia conditioned responses (CRs) to tonal CSs. Whereas the bradycardia responses to the CS+ quickly extinguished in the group that had control lesions in the visual cortex, the auditory cortex lesion group continued to exhibit significantly larger bradycardiac HR CRs to the CS+ relative to the CS− during all 7 days of extinction. These results suggest that the animals in the auditory cortex lesioned group did not inhibit responses to a previously reinforced stimulus (i.e. CS+) as well as animals with control lesions in the visual cortex.     

Dorsal striatum and stimulus-response learning: lesions of the dorsolateral, but not dorsomedial, striatum impair acquisition of a simple discrimination task

In the present experiment, the effects of neurotoxic lesions (quinolinic acid) of the dorsolateral or dorsomedial striatum were investigated on a simple instrumental discrimination task (CS+/CS−). Rats with lesions of the dorsolateral striatum were found to be impaired in the acquisition of this task, as compared to rats with either dorsomedial striatal or sham lesions. Furthermore, dorsolateral striatal lesioned animals had significantly lower levels of responding across the course of discrimination training, as assessed both by overall rate of response during CS+ presentations and number of CS+ trials without a response, despite having shown levels of responding during variable interval training that did not differ from that of sham lesioned animals. In contrast, animals with lesions of the dorsomedial striatum did not show an impairment in acquisition of the present task, but had slightly higher rates of responding during CS− presentations. It is argued that the poor acquisition and low response rates observed in animals with dorsolateral striatal lesions reflect a failure in stimulus–response learning, while the performance of animals with dorsomedial striatal lesions may have been the result of an increase in overall activity rate.     

Retention of conditioned inhibition of the nictitating membrane response in decorticate rabbits

Rabbits were trained on a Pavlovian conditioned inhibition (CI) task using light as the reinforced conditional stimulus (CS) and the same light compounded with a tone as the non-reinforced CS. The conditional response was the nictitating membrane response (NMR). The subjects then received an extensive neocortical lesion or a sham operation. After 9 weeks postoperative recovery, the animals were retrained on the CI task. Sham operated control animals showed immediate and high levels of CI retention but the decorticates showed a profound initial loss, showing that there is a cortical influence in CI. Reacquisition of CI by the decorticates was rapid and showed considerable savings over de novo acquisition. This supports our earlier finding that the neocortex is not essential for the acquisition of CI.     

Hippocampal lesions prevent trace eyeblink conditioning in the freely moving rat.

The effect of hippocampal aspiration lesions on trace eyeblink conditioning was examined in young, freely-moving F1 hybrid rats (Fisher 344 x Brown Norway). Rats which received either bilateral neocortical or bilateral hippocampal aspiration lesions were compared with each other or with sham lesioned control rats. The rats were trained with a 250 ms tone conditioning stimulus (CS), a 250 ms stimulus free trace interval and a 100 ms corneal airpuff unconditioned stimulus (US). Rats with lesions of the hippocampus were significantly impaired relative to the neocortical and sham lesioned control rats. Analyses of different behavioral parameters (e.g. percent conditioned responses, amplitude, and area of response) indicated that all of the measures for the conditioned response were significantly impaired by the hippocampal lesion. The unconditioned response was not significantly affected by the lesion, and there was no significant difference among the groups after 2 days of subsequent conditioning with the delay paradigm (zero trace interval). We conclude that the hippocampus is required for rats to learn the association between a tone CS and an airpuff US when a 250 ms trace interval is interposed between the two stimuli.     

A thalamo-cortico-amygdala pathway mediates auditory fear conditioning in the intact brain

The neural substrates of fear conditioning in rats have been well characterized, with converging lines of evidence indicating that conditioned stimulus (CS) and unconditioned stimulus (US) information form a CS-US association in the amygdala. Auditory CS information can reach the amygdala via two routes: a direct thalamo-amygdala pathway, and an indirect thalamo-cortico-amygdala pathway. Although either pathway can fully support learning when the alternate pathway is disrupted, many studies to date have argued that the thalamo-amygdala pathway is the principal auditory CS pathway in intact brains. To test this hypothesis, we trained rats in auditory fear conditioning, and 24 h later lesioned either pathway, leaving the alternate pathway intact. Later, animals were tested for conditioned freezing to the auditory CS. We report that lesions of the thalamo-amygdala pathway produced severe but incomplete deficits in freezing during the tone retention test, while lesions of the thalamo-cortico-amygdala pathway completely abolished freezing during tone presentation. These results suggest that the thalamo-cortico-amygdala pathway is the principal auditory CS pathway when the brain is intact.