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Electrolytic lesions to nucleus accumbens core and shell have dissociable effects on conditioning to discrete and contextual cues in aversive and appetitive procedures respectively.

The nucleus accumbens (n. acc.) has been implicated in conditioning to both discrete and contextual cues but its precise role is as yet controversial because conflicting patterns of effect have been reported. These inconsistencies may relate to the extent to which the lesions used encroach on different subfields of n. acc. and the use of different task variants. The present study compared the effects of selective lesions of shell and core subfields of nucleus accumbens (n. acc.) across aversive and appetitive trace conditioning variants. In both experiments, an auditory stimulus was contiguous with footshock or food, or presented at a trace interval. A continuous flashing light in each case provided an experimental background stimulus. Conditioning to the cues provided by the experimental chambers was also assessed. Rats with electrolytic lesions to the n. acc. shell and core showed different patterns of effect in aversive (Experiment 1) and appetitive (Experiment 2) variants of this procedure. In Experiment 1, the core lesion reduced the difference between trace and contiguously conditioned groups, in responding to the discrete noise stimulus. However, neither lesion had any detectable effect on contextual conditioning. In Experiment 2, the shell lesion clearly increased contextual conditioning, selectively in the trace conditioned group, but neither lesion had any detectable effect on discrete cue conditioning. Thus, whilst the shell and core lesions produced dissociable effects on discrete cue and contextual conditioning, the conclusions to be drawn depend on the procedural variant in use.

Animals↗

Repeated ethanol exposure and withdrawal impairs human fear conditioning and depresses long-term potentiation in rat amygdala and hippocampus.

BACKGROUND: In rats, repeated episodes of alcohol consumption and withdrawal (RWD) impair fear conditioning to discrete cues. METHODS: Fear conditioning was measured in human binge drinkers as the increased startle response in the presence of a CS+ conditioned to aversive white noise. Secondly, the ability of tone CSs, paired with footshock, to induce c-fos expression, a marker of neuronal activity, in limbic structures subserving emotion was studied in rats. Additionally, consequences of RWD on subsequent induction of long term potentiation (LTP) in external capsule/lateral amygdala and Schaffer collateral/hippocampus CA1 pathways were studied in rat brain slices. RESULTS: Fear conditioning was impaired in young human binge drinkers. The ability of fear-conditioned CSs to increase c-fos expression in limbic brain areas was reduced following RWD, as was LTP induction. Rats conditioned prior to RWD, following RWD showed generalization of conditioned fear from the tone CS+ to a neutral control stimulus, and a novel tone. CONCLUSIONS: Binge-like drinking impairs fear conditioning, reduces LTP, and results in inappropriate generalization of learned fear responses. We propose a mechanism whereby RWD-induced synaptic plasticity reduces capacity for future learning, while allowing unconditioned stimuli access to neuronal pathways underlying conditioned fear.

Adult↗

Cerebellar activation during leg withdrawal reflex conditioning: an fMRI study.

OBJECTIVE: The aim of the present study was to examine cerebellar areas related to conditioning of the nociceptive leg withdrawal reflex using event-related functional magnetic resonance imaging (fMRI). Because of the aversive nature of the unconditioned stimulus effects of accompanying fear conditioning were expected. METHODS: In 20 healthy adult subjects leg withdrawal reflex conditioning was performed using a standard delay protocol during MR-scanning. Electromyographic recordings from the anterior tibial and biceps femoris muscles were used to quantify conditioned responses. Fear-related changes of heart rate were assessed. RESULTS: In the group of all subjects a significant increase of cerebellar activation was found in the anterior and posterior vermis. In the group of subjects (n=9) who showed conditioned leg withdrawal responses cerebellar activation was more pronounced in parts of the anterior vermis, which correspond to the known leg representation. In the group of subjects (n=11) who did not develop conditioned responses cerebellar activation was more pronounced in the posterolateral hemispheres. Changes of heart rate, however, did not significantly differ between groups. CONCLUSIONS: Results suggest that areas within the anterior vermis are involved in conditioning of the leg withdrawal response. The present results, however, do not allow to differentiate between motor performance, learning or timing-related processes. Areas in the posterior vermis and cerebellar hemispheres may be related to concomitant fear conditioning. SIGNIFICANCE: Results of the present event-related fMRI study suggest involvement of the human cerebellum in conditioning of the nociceptive leg withdrawal response.

Adult↗

Time course of amygdala activation during aversive conditioning depends on attention.

The time course of amygdala activation during aversive conditioning is a matter of debate. While some researchers reported rapid habituation, others found stable or no amygdalar responses to conditioned stimuli at all. In the present event-related fMRI study, we investigated whether the activity of the amygdala during aversive conditioning depends on attentional conditions. Subjects underwent aversive delay conditioning by pairing an electrical shock (unconditioned aversive stimulus) with a visual conditioned stimulus (CS+). For each singular presentation of the CS+ or a nonconditioned visual stimulus (CS-), subjects attended in random order to features that either differed between both stimuli (identification task) or that did not differ (distraction task). For the identification task trials, increased responses of the left amygdala to CS+ versus CS- were rapidly established but absent at the end of the conditioning trials. In contrast, under the distraction condition, amygdala activation to CS+ versus CS- was present during the late but not the early phase of conditioning. The results suggest that the time course of amygdala activity during aversive associative learning is strongly modulated by an interaction of attention and time.

Adolescent↗

Overt behavior and ultrasonic vocalization in a fear conditioning paradigm: a dose-response study in the rat.

The behavioral analysis of laboratory rats is usually confined to the level of overt behavior, like locomotion, behavioral inhibition, instrumental responses, and others. Apart from such visible outcome, however, behaviorally relevant information can also be obtained when analyzing the animals' ultrasonic vocalization, which is typically emitted in highly motivational situations, like 22-kHz calls in response to acute or conditioned threat. To further investigate such vocalizations and their relationship with overt behavior, we tested male Wistar rats in a paradigm of Pavlovian fear conditioning, where a tone stimulus (CS) was preceding an aversive foot-shock (US) in a distinct environment. Importantly, the shock dose was varied between groups (0-1.1 mA), and its acute and conditioned outcome were determined. The analysis of visible behavior confirms the usefulness of immobility as a measure of fear conditioning, especially when higher shock doses were used. Rearing and grooming, on the other hand, were more useful to detect conditioned effects with lower shock levels. Ultrasonic vocalization occurred less consistently than changes in overt behavior; however, dose-response relationships were also observed during the phase of conditioning, for example, in latency, call rate and lengths, intervals between calls, and sound amplitude. Furthermore, total calling time (and rate) were highly correlated with overt behavior, namely behavioral inhibition as measured through immobility. These correlations were observed during the phase of fear conditioning, and the subsequent tests. Importantly, conditioned effects in overt behavior were observed, both, to the context and to the CS presented in this context, whereas conditioned vocalization to the context was not observed (except for one rat). In support and extent of previous results, the present data show that a detailed analysis of ultrasonic vocalization can substantially broaden and refine the spectrum of analysis in behavioral work with rats, since it can provide information about situational-, state-, and subject-dependent factors which are partly distinct from what is visible to the experimenter.

Acoustic Stimulation↗

Acid taste thresholds assessed by conditioned taste aversion and two-bottle preference in rats.

The conditioned taste aversion (CTA) threshold for either citric acid (CA) or HCl solutions and the two-bottle taste preference (TBP) threshold were determined in rats that are familiarized to the odor of conditioning solutions or that are naive. The CTA method appeared to be more sensitive than the TBP test, particularly when rats were not familiarized to the odor of the conditioning solution. The CTA threshold for HCl-conditioned rats and familiarized to the odor of conditioning solution lies between 1.00 and 2.00 mmol; in unconditioned rats, it lies between 4.00 and 5.00 mmol. In CA-conditioned and odor-familiarized rats, the threshold lies between 0.09 and 0.20 mmol; in unconditioned rats, it lies between 7.00 and 10.00 mmol. In rats not familiarized to the odor of the conditioning solution, the threshold for HCl-conditioned rats lies between 0.90 and 1.00 mmol; in unconditioned rats, it lies between 2.00 and 3.00 mmol. In CA-conditioned rats, the CTA threshold lies between 0.03 and 0.05 mmol; in unconditioned rats, it lies between 4.00 and 7.00 mmol. The two-bottle test is less sensitive than the CTA method. The TBP threshold lies between HCl 4.00 and 5.00 mmol, and between CA 4.00 and 7.00 mmol. The odor of a solution may potentiate the ability of rats to detect the concentration of CA and HCl solutions.

Animals↗

Influence of in-tunnel environment to in-bus air quality and thermal condition in Hong Kong.

In this study, the potential exposure of bus commuters to significant air parameters (CO(2), CO and RSP) and thermal environment (air temperature and relative humidity) when buses traveled through tunnels in Hong Kong was investigated. It was found that air-conditioned buses provided a better commuting environment than non-air-conditioned buses. The blate increasing trend was found on air-conditioned buses as the in-bus air parameters concentration levels rose slowly throughout the traveling process. In contrast, the in-bus environment varied rapidly on non-air-conditioned buses as it depended on the out-bus environment. The measured in-bus CO concentration was 2.9 ppm on air-conditioned buses, while it was 4.6 ppm (even reaching the highest level at 12.0 ppm) on non-air-conditioned buses. Considering the in-bus thermal environment, air-conditioned buses provided thermally comfortable cabins (about 24 degrees C and 59% of relative humidity). However, on non-air-conditioned buses, the thermal environment varied with the out-bus environment. The mean in-bus air temperature was about 34 degrees C and 66% of relative humidity, and the in-bus air temperature varied between 29 and 38 degrees C. Also, the lower-deck to upper-deck air parameters concentration ratios indicated that the vertical dispersion of air pollutants in tunnels influenced non-air-conditioned buses as higher air parameters concentration levels were obtained on the lower-deck cabins.

Air Conditioning↗

The human cerebellum and associative learning: dissociation between the acquisition, retention and extinction of conditioned eyeblinks.

The present paper is part of a systematic exploration of the neural substrates of conditioned eyeblink responses in humans. Normal subjects and patients with lesions restricted to the cerebellum were examined for their ability to acquire new classically conditioned eyeblinks to an auditory conditioned stimulus and whether they were able to perform and extinguish a previously learned natural anticipatory eyeblink response - the kinesthetic threat eyeblink response (KTER). In classical conditioning to an auditory conditioned stimulus, cerebellar patients failed to acquire new conditioned responses. In contrast to this impairment, in the KTER task both cerebellar patients and control subjects exhibited a high incidence of anticipatory eyeblinks which were initiated before the forehead tap. These results indicate that the cerebellar circuits, which are critical for the acquisition of new conditioned responses, are not essential for the storage and expression of naturally acquired conditioned responses. In the extinction experiment, cerebellar patients failed to extinguish their KTERs. This finding suggests that in humans, the acquisition of new and the extinction of previously learned conditioned responses depends on a similar set of cerebellar circuits.

Animals↗

Sulpiride injections into the medial septum reverse the influence of intra-medial septum injection of L-arginine on expression of place conditioning-induced by morphine in rats.

Effects of intra-medial septum injections of L-arginine, a precursor of nitric oxide, N(G)-nitro-L-arginine methyl ester (L-NAME), an inhibitor of nitric oxide synthase, and sulpiride, a selective antagonist of dopamine D2 receptor on morphine-induced conditioned place preference (CPP) in male Wistar rats were examined. Using a 3-day schedule of conditioning, morphine (0.5-7.5 mg/kg, s.c.) produced a significant place preference in a dose-dependent manner. The maximum response was observed with 5.0 mg/kg of opioid. Sulpiride (0.3, 1.0 and 3.0 microg/rat), but not L-arginine (0.3, 1.0 and 3.0 microg/rat) or L-NAME (0.3, 1.0 and 3.0 microg/rat), in combination with morphine (5.0 mg/kg), during conditioning, significantly altered morphine-induced CPP. Single doses (0.3, 1.0 and 3.0 microg/rat) of either L-arginine or L-NAME, during conditioning, did not induce CPP. Sulpiride at 0.3-3.0 microg/rat, intra-medial septum, during conditioning, produced a significant conditioned place aversion. Intra-medial septum injections of L-arginine but not L-NAME or sulpiride, 1-2 min before testing, increased the expression of morphine-induced CPP. The administration of sulpiride (0.3, 1.0 and 3.0 microg/rat), but not L-NAME (0.3, 1.0 and 3.0 microg/rat), 1-2 min before the injection of L-arginine (0.3 microg/rat) on day of test, significantly attenuated the response to L-arginine. L-Arginine (0.3-3.0 microg/rat), during conditioning, showed a statistically significant increase in locomotor activity compared with that to control group. Moreover, sulpiride decreased locomotion by itself or in combination with morphine during conditioning and on the test day of morphine CPP. It can be concluded that L-arginine, a precursor of nitric oxide, in the rat median septum may play a role in expression of morphine conditioning due to dopamine release in this area.

Animals↗

Hippocampus in delay eyeblink classical conditioning: essential for nefiracetam amelioration of learning in older rabbits.

Rabbits acquire conditioned responses (CRs) normally with bilateral removal of the hippocampus, but alterations of the intact hippocampus can affect the rate of acquisition. The cognition-enhancing drug, nefiracetam ameliorated the acquisition of CRs in older rabbits, protected membrane dysfunction in hippocampal CA1 neurons following oxygen and glucose deprivation, and promoted the release of diverse neurotransmitters, including acetylcholine. Because the septo-hippocampal cholinergic system is demonstrated to be involved in eyeblink conditioning, this experiment was undertaken to explore whether nefiracetam ameliorates conditioning via the hippocampus. Data from 53 rabbits of a mean age of 28 months were tested under two drug conditions (10 or 0 mg/kg nefiracetam) and 4 lesion conditions (bilateral hippocampectomy, bilateral neocortical removal, sham surgery, no surgery). The three groups of nefiracetam-treated rabbits with intact hippocampus acquired CRs more rapidly than the vehicle-treated groups, but rabbits with bilateral hippocampectomy treated with nefiracetam learned like vehicle-treated rabbits. Results suggest that nefiracetam ameliorates learning via the hippocampus. Because of the parallels between conditioning in rabbits with disrupted hippocampal cholinergic systems and conditioning in Alzheimer's disease (AD), these results suggest that nefiracetam may ameliorate conditioning in AD as it ameliorates conditioning in older rabbits.

Aging↗

A role for brain glucocorticoid receptors in contextual fear conditioning: dependence upon training intensity.

We studied the possible involvement of corticosteroids in the establishment and long-term expression of contextual fear conditioning and questioned whether a corticosteroid action might be dependent upon stimulus intensity at training. Experiments included: (i) the intracerebroventricular administration of specific antagonists for the two types of intracellular corticosteroid receptors to rats trained at either 1 mA or 0.4 mA shock intensity at conditioning; and (ii) the administration of corticosterone after conditioning rats to 0.2 mA shocks. The results showed that the administration of a type II glucocorticoid, but not a type I mineralocorticoid, receptor antagonist before conditioning rats to the intermediate shock condition attenuated long-term expression of contextual fear conditioning. However, treatment with the antagonists before conditioning to the high shock intensity failed to influence the extent of fear conditioning. In addition, an intraperitoneal corticosterone injection, given immediately after training rats at the low shock intensity, enhanced long-term expression of the fear response. The results support the view that post-training levels of circulating corticosterone, through an interaction with central type II glucocorticoid receptors, modulate the strength to which memory for contextual fear conditioning is established and maintained.

Animals↗

Influence of nitric oxide on morphine-induced conditioned place preference in the rat central amygdala.

Effects of intra-central amygdala injections of L-arginine, a nitric oxide (NO) precursor, and N(G)-nitro-L-arginine methyl ester (L-NAME), a NO synthase (NOS) inhibitor, on morphine-induced conditioned place preference in rats were investigated by using an unbiased 3-day schedule of place conditioning design. Animals receiving once daily injections of morphine (0.5-7.5 mg/kg, subcutaneously, s.c.) or saline (1.0 ml/kg, s.c.) showed a significant place preference in a dose-dependent manner. The maximum response was observed with 5.0 mg/kg of the opioid. Co-administration of morphine (5.0 mg/kg) with L-arginine (0.3, 1.0 and 3.0 microg/rat), but not with L-NAME (0.3, 1.0 and 3.0 microg/rat), during the acquisition of morphine-induced conditioned place preference increased morphine-induced conditioned place preference. The response to L-arginine was blocked by L-NAME preadministration. L-arginine and L-NAME by themselves did not induce conditioned place preference. When L-arginine or L-NAME at 0.3-3.0 microg/rat was administered 1 min before conditioned place preference testing, L-arginine but not L-NAME caused an increase in the expression of morphine-induced conditioned place preference, the effect that was blocked by L-NAME preadministration. A dose of L-arginine (0.3 microg/rat), but not L-NAME, during expression of morphine-induced conditioned place preference produced an increase in locomotion compared with that in the control group. It may be concluded that an increase in the NO levels in the central amygdala may have an effect on the acquisition and expression of morphine-induced conditioned place preference.

Amygdala↗

5-HT mechanisms of median raphe nucleus in the conditioned freezing caused by light/foot-shock association.

We have shown that 5-HT mechanisms of the median raphe nucleus (MRN) are involved in contextual fear-conditioning processes as electrolytic or neurotoxic lesions with N-methyl-D-aspartate (NMDA) or injections of 8-hydroxy-2-(di-n-propilamino)-tetralin (8-OH-DPAT) into this structure inhibit freezing behavior in a contextual fear paradigm. In this work, we extend these studies by analyzing the behavioral responses in a classical fear-conditioning paradigm (light or tone/foot-shock association) in rats with either neurochemical lesion with NMDA or injected with 8-OH-DPAT into the MRN. The animals received NMDA or 8-OH-DPAT or saline microinjections into the MRN and were submitted to conditioning trials in an experimental chamber, where they received 10 foot-shocks (0.6 mA, 1 s, variable interval between 10 and 50 s) paired with tone or light (CS). On the next day, they were tested in a different experimental chamber, with or without CS presentation, where the duration of freezing and the number of rearing episodes were recorded. Light or tone alone caused a significant amount of freezing. NMDA lesions or 8-OH-DPAT injections into the MRN clearly inhibited freezing behavior in rats conditioned to light/foot-shock association, but not in the conditioning sessions with tones. Besides the proposed role in contextual fear conditioning, these results clearly show that MRN is involved in the fear conditioning with light as conditioned stimuli. Distinct neural substrates seem to subserve conditioning fear with acoustic stimuli.

8-Hydroxy-2-(di-n-propylamino)tetralin↗

Hippocampal acetylcholine increases during eyeblink conditioning in the rabbit.

The classically conditioned rabbit nictitating membrane reflex (NMR) is modulated by the septohippocampal cholinergic system. Disruption of this system retards NMR acquisition. Aluminium (Al) is a neurotoxin that interferes with hippocampal acetylcholine (ACh) synthesis and release. Using microdialysis, this study tested the hypothesis that NMR acquisition in the rabbit is associated with hippocampal ACh release. This was conducted by measuring ACh release in control and A1-intoxicated rabbits during NMR training. NMR training consisted of four sessions of 100 conditioning trials/session in a delay paradigm. The percentage of conditioned responses (CRs) increased with each conditioning session for both groups, although percent CRs was significantly greater in the control group. Acetylcholine release in the ventral hippocampus increased significantly over baseline in the control group during the second and third conditioning sessions. In the Al-intoxicated group, ACh release did not increase significantly during any conditioning session. A separate group of rabbits was pseudoconditioned, receiving the same conditioning stimuli, although explicitly unpaired. This group did not acquire the CR. Acetylcholine release did not significantly increase during any conditioning session, suggesting that the increase in ACh release observed in the control group was not merely a product of conditioning stimuli presentation. The lack of increased ACh release in the Al-intoxicated rabbits was associated with a CR acquisition deficit. The results of this study are consistent with a role of hippocampal cholinergic function in NMR acquisition in the rabbit.

Acetylcholine↗

Conditioned insulin and blood sugar responses in humans in relation to binge eating.

This study proposed to demonstrate a classically conditioned blood sugar decrease in humans and to clarify its relevance for binge eating. Six conditioning trials were run in healthy females. The conditioned stimulus (CS) was a compound peppermint flavor/fragrance, whereas the unconditioned stimulus (UCS) consisted of 50 g of oral glucose. Control subjects received an aspartame drink as the UCS. Ad lib glucose intake, blood parameters, and subjective craving were monitored before and after conditioning. Results showed that the experimental group failed to show conditioned blood sugar and glucagon decreases or C-Peptide increases. Although an increased insulin response was found in the experimental group, the effect size did not exceed that of spontaneous fluctuations. No increases in craving for sweet substances were found. An impressive increase (mean: 78%) in glucose intake after conditioning was found in both conditions, as well as in a subsequently run third condition with plain water as the UCS. The increased glucose intake probably resulted from an initial neophobia to the laboratory setting that subsided as subjects had experienced more lab sessions. Importantly, because no conditioned hypoglycemia occurred in the present study, its relationship with subjectively experienced craving for sweet substance could not be determined.

Adolescent↗

Effect of the dopamine D(1/5) antagonist SCH 23390 on the acquisition of conditioned fear.

The authors previously reported that typical and atypical antipsychotic drugs inhibited the acquisition but not expression of conditioned fear. The present study examined the effects of the selective dopamine D(1/5) agonist (SKF 38393) and antagonist (SCH 23390) on the acquisition and expression of conditioned fear. Drugs were administered subcutaneously to male Sprague-Dawley rats 30 min before foot shock (2.5 mA for 5 min). Twenty-four hours after foot shock, rats were again placed and observed in the shock chamber without shocks (conditioned fear). Freezing behavior induced by conditioned fear, an index of anxiety or fear, was recorded using a time-sampling procedure. SCH 23390 (0.1-1 mg/kg) inhibited the acquisition of conditioned freezing. The administration of SCH 23390 at a dose of 0.1 mg/kg 30 min after foot shock did not affect conditioned freezing. Taken together, it is concluded that D(1/5) antagonism inhibits the acquisition of conditioned fear. SKF 38393 (3-20 mg/kg) failed to change the acquisition of conditioned fear. SCH 23390 or SKF 38393 administered prior to testing did not reduce the expression of conditioned fear. These results suggest that D(1/5) receptors may play a role in the development of fear or anxiety.

Animals↗

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.

Acoustic Stimulation↗

Amygdala, hippocampus and discriminative fear conditioning to context.

Various measures of fear have been shown to condition to a fearful context with different acquisition rates (Antoniadis EA, McDonald RJ. Fear conditioning to context expressed by multiple measures of fear in the rat, Behav Brain Res 1999;101(1):1-14). Freezing, locomotion, urination and preference are 'fast' measures of fear in that they discriminatively condition to context after a single training session, while ultrasonic vocalizations and defecation are 'slow' measures of fear given that they condition following three training sessions. In the present experiment we sought to assess the contribution of the amygdala and the hippocampus in this form of learning. Existing views differ on the degree of involvement of each memory structure. This discord probably emerges from the common use of non-discriminative paradigms and the assessment of a single measure of fear. With the use of a discriminative paradigm and the assessment of multiple measures of fear, results indicate that the amygdala is a memory structure that selectively mediates the conditioning of heart rate, and the hippocampus selectively mediates the conditioning of defecation and body temperature. The conditioning of preference, locomotion, freezing and ultrasonic vocalizations, necessitate the participation of both memory structures while the conditioning of urination does not seem to require the participation of either the hippocampus or the amygdala. The proposed view ascribes an equal role in fear conditioning to both the amygdala and the hippocampus.

Amygdala↗