Footshock Intensity Research Articles

Cycloheximide impairs and enhances memory depending on dose and footshock intensity

This experiment examined the effects on memory of interactions of cycloheximide dose and training foot shock intensity. Mice received injections of cycloheximide (120mg/kg, s.c.) or saline 30min prior to inhibitory avoidance training with shock intensities of 100, 150, 250 or 300μA (1s duration). Memory was tested 48h later. The saline control mice showed increasing memory latencies as a function of shock intensity. The ability of cycloheximide to impair memory increased as the training shock intensity increased. In a second experiment, mice were trained with a 200μA (1s duration) shock and received injections of saline or cycloheximide at one of several doses (30, 60 or 120mg/kg). Under these training conditions, cycloheximide enhanced memory in an inverted-U dose–response manner. These findings are consistent with prior findings suggesting that protein synthesis inhibitors act on memory by altering modulators of memory formation as a secondary consequence of the inhibition of protein synthesis rather than by interfering with training-initiated synthesis of proteins required for memory formation.

Re-examining the ontogeny of the context preexposure facilitation effect in the rat through multiple dependent variables

The capability to acquire context conditioning does not emerge until weaning, at least when the defining features of the context lack explicit and salient olfactory cues. Contextual learning deficits in preweanling rats have been associated with functional immaturity of the dorsal hippocampus. According to recent studies, the so-called context preexposure facilitation effect (CPFE) – a hippocampus-dependent effect – is not observed until postnatal day 23 (PD23). In these studies the footshock intensity employed was higher (1.5mA) than in adult studies, and context conditioning was inferred from a single behavioral measure (percentage of freezing). The present study examined the CPFE on PD17 and PD23 by analyzing multiple dependent variables, including fecal boli and an ethogram covering the complete behavioral repertoire of the rat. A non-shocked control group was included in the design and two footshock intensities were employed (0.5 and 1.5mA). Results showed clear evidence of contextual fear conditioning in preweanling and weanling rats, as well as evidence of conditioned fear in non-preexposed rats from both age groups. In some cases, some dependent variables, such as grooming or vertical exploration, were more sensitive than freezing for detecting evidence of memory. Strong fear responses were detected in weanling (but not preweanling) rats, when rats were evaluated in a different context from the one employed at conditioning. These results indicate that preweanling rats are capable of acquiring contextual conditioning, even in a context lacking explicit odor cues, and highlight the importance of multiple dependent variables for analyzing the ontogeny of memory.

Extinction procedure induces pruning of dendritic spines in CA1 hippocampal field depending on strength of training in rats.

Numerous reports indicate that learning and memory of conditioned responses are accompanied by genesis of dendritic spines in the hippocampus, although there is a conspicuous lack of information regarding spine modifications after behavioral extinction. There is ample evidence that treatments that typically produce amnesia become innocuous when animals are submitted to a procedure of enhanced training. We now report that extinction of inhibitory avoidance (IA), trained with relatively low foot-shock intensities, induces pruning of dendritic spines along the length of the apical dendrites of hippocampal CA1 neurons. When animals are trained with a relatively high foot-shock there is a high resistance to extinction, and pruning in the proximal and medial segments of the apical dendrite are seen, while spine count in the distal dendrite remains normal. These results indicate that pruning is involved in behavioral extinction, while maintenance of spines is a probable mechanism that mediates the protecting effect against amnesic treatments produced by enhanced training.

Early fear as a predictor of avoidance in a rat model of post-traumatic stress disorder

Exposure of humans and animals to an intensely fearful experience can lead to an enduring behavioral profile involving fear and avoidance. The present study examined if rats that show more fear to a novel tone one day after exposure to footshocks exhibit more avoidance-like responses over a 4-week period. Rats were exposed to an episode of moderately intense footshock (5 × 2 s episodes of 1.5 mA presented randomly over 3 min). Shock rats that exhibited a high level of fear (HR) to a novel tone one day after the shock exposure showed more avoidance of open spaces and novel rats when compared to shock rats that exhibited a lower level of fear to the novel tone (LR). Similarly, HR emitted more ultrasonic vocalization in the dysphoric range (20–30 kHz) when placed in a novel chamber or the chamber in which shock was given. This study highlights the importance of early fear as a contributing factor for the development of lasting changes in avoidance. These results also support the view that the presence of an intense peritraumatic stress response may be a predictor of the subsequent development of a lasting negative emotional state in humans exposed to trauma.

Motivation for palatable food despite consequences in an animal model of binge eating

Binge eating involves an abnormal motivation for highly palatable food in that these foods are repeatedly consumed despite their binge-triggering effects and life-affecting consequences associated with binge eating. We determined if rats identified as binge-eating prone (BEP) similarly display abnormal motivation for palatable food. Food-sated BEP and binge-eating resistant (BER) rats were given voluntary access to palatable food paired with increasing intensity of footshock. Later, they were exposed to a period of cyclic caloric restriction-refeeding. BEPs consumed significantly more and tolerated higher levels of footshock for palatable food than BERs. Cyclic restriction-refeeding increased BERs' tolerance of shock for palatable food. Previously observed parallels of the rat BEP model to human binge eating can now be extended to include an abnormal motivation for palatable food. This model should prove useful in identifying specific genes that interact with the nutritional environment to mediate binge eating and may point to novel physiological targets to treat compulsive overeating.

Avoidance perseveration during extinction training in Wistar-Kyoto rats: An interaction of innate vulnerability and stressor intensity

Given that avoidance is a core feature of anxiety disorders, Wistar-Kyoto (WKY) rats may be a good model of anxiety vulnerability for their hypersensitivity to stress and trait behavioral inhibition. Here, we examined the influence of strain and shock intensity on avoidance acquisition and extinction. Accordingly, we trained WKY and Sprague-Dawley (SD) rats in lever-press avoidance using either 1.0-mA or 2.0-mA foot-shock. After extinction, neuronal activation was visualized by c-Fos for overall activity and parvalbumin immunoreactivity for gamma-aminobutyric acid (GABA) neuron in brain areas linked to anxiety (medial prefrontal cortex and amygdala). Consistent with earlier work, WKY rats acquired lever-press avoidance faster and to a greater extent than SD rats. However, the intensity of foot shock did not differentially affect acquisition. Although there were no differences during extinction in SD rats, avoidance responses of WKY rats trained with the higher foot shock perseverated during extinction compared to those WKY rats trained with lower foot shock intensity or SD rats. WKY rats trained with 2.0-mA shock exhibited less GABAergic activation in the basolateral amygdala after extinction. These findings suggest that inhibitory modulation in amygdala is important to ensure successful extinction learning. Deficits in avoidance extinction secondary to lower GABAergic activation in baslolateral amygdala may contribute to anxiety vulnerability in this animal model of inhibited temperament.

Repeated exposure to immobilization or two different footshock intensities reveals differential adaptation of the hypothalamic–pituitary–adrenal axis

Factors involved in adaptation to repeated stress are not well-characterized. For instance, acute footshock (FS) of high intensity appears to be less severe than immobilization (IMO) in light of the speed of post-stress recovery of the hypothalamic-pituitary-adrenal (HPA) axis and other physiological variables. However, repeated exposure to IMO consistently resulted in reduction of the HPA response to the same stressor (adaptation), whereas failure to adapt has been usually reported after FS. Thus, in the present work we directly compared the activation of HPA axis and other physiological changes in response to both acute and repeated exposure to IMO and two intensities of FS (medium and high) in adult male rats. Control rats were exposed to the FS boxes but they did not receive shocks. Daily repeated exposure to IMO resulted in significant adaptation of the overall ACTH and corticosterone responses to the stressor. Such a reduction was also observed with repeated exposure to FS boxes and FS-medium, whereas repeated exposure to FS-high only resulted in a small reduction of the corticosterone response during the post-stress period. This suggests that some properties of FS-high make adaptation to it difficult. Interestingly, overall changes in food intake and body weight gain throughout the week of exposure to the stressors reveal a greater impact of IMO than FS-high, indicating that factors other than the intensity of a stressor, at least when evaluated in function of the above physiological variables, can influence HPA adaptation. Since FS exposure is likely to cause more pain than IMO, activation of nociceptive signals above a certain level may negatively affect HPA adaptation to repeated stressors.

Enhanced training protects memory against amnesia produced by concurrent inactivation of amygdala and striatum, amygdala and substantia nigra, or striatum and substantia nigra

Memory is markedly impaired when normal activity of any of a number of cerebral structures is disturbed after a learning experience. A growing body of evidence indicates, however, that such interference with neuronal function becomes negligible when the learning experience is significantly enhanced. We now report on the effects of enhanced training on retention after temporary inactivation of cerebral nuclei known to be involved in memory, namely the substantia nigra (SN), striatum (STR), and amygdala (AMY). When training was conducted with a relatively low intensity of footshock (1.0 mA), post-training infusion of lidocaine into the SN, STR, or AMY produced a marked memory deficit. Increasing the aversive stimulation to 2.0 mA protected memory from the amnesic effect of intranigral lidocaine, but there was still a deficit after its infusion into the STR and AMY. Administration of lidocaine into each of these nuclei, in the groups that had been trained with 3.0 mA, was completely ineffective in producing alterations in memory consolidation. Simultaneous infusion of lidocaine into STR + SN, AMY + SN, or AMY + STR was also ineffective in altering memory formation when the highest footshock intensity was used for training. To our knowledge, this is the first demonstration that an enhanced learning experience guards against memory deficits after simultaneous temporary interruption of neural activity of brain nuclei heretofore thought to be necessary for memory formation. These findings support the proposition that brain structures involved in memory processing are functionally connected in series during memory consolidation and that, after an enhanced learning experience, these structures become functionally connected in parallel.

Hippocampal alpha7 nicotinic receptors modulate memory reconsolidation of an inhibitory avoidance task in mice

CF-1 male mice were trained in an inhibitory avoidance (IA) task using either a mild or a high footshock (0.8 or 1.2 mA, 50 Hz, 1 s). A retention test was given 48 h later. Immediately after the retention test, mice were given intra-dorsal hippocampus infusions of either choline (Ch, an α7 nicotinic acetylcholine receptor (α7nAChR) agonist, 0.08–1.30 μg/hippocampus), or methyllycaconitine (MLA, an α7nAChR antagonist, 1.0–30.0 μg/hippocampus). Memory retention was tested again 24 h later. Methyllycaconitine impaired retention performance regardless of footshock intensity and its effects were long lasting. Ch impaired retention performance only in those mice trained with a high footshock. On the contrary, Ch enhanced retention performance when mice were trained with a mild footshock. These effects were long lasting and dose- and time-dependent. Retention performance was not affected in drug-treated mice that were not subjected to memory reactivation, suggesting that the performance effects could not be attributable to non-specific effects of the drugs. Methyllycaconitine effects were dose-dependently reversed by choline, suggesting that MLA and Ch interact at the α7nAChR. Altogether, results suggest that hippocampal α7nAChRs play a critical role in reconsolidation of an IA response in mice, and may also have important implications for dynamic memory processes. This is the first presentation, to our knowledge, indicating that a specific receptor (α7nAChR) is able to modulate consolidated memories after retrieval.

Limited efficacy of propranolol on the reconsolidation of fear memories

Previous studies suggested that the beta-adrenergic receptor antagonist propranolol might be a novel, potential treatment for post-traumatic stress disorder (PTSD). This hypothesis stemmed mainly from rodent studies showing that propranolol interferes with the reconsolidation of Pavlovian fear conditioning (FC). However, subsequent investigations in humans have produced controversial evidence about the effect of propranolol on fear memories and an effect on PTSD symptomatology has yet to be reported. Thus, it remains to be established whether propranolol interferes with the reconsolidation of fear memories at large. To address this question, we tested the effect of systemic injections of propranolol administered before or after the retrieval of an inhibitory avoidance (IA) memory elicited with different footshock intensities. In parallel, the same treatment was tested on the reconsolidation of Pavlovian FC. Propranolol showed no effect on the reconsolidation of IA, although the pre-retrieval administration resulted in a significant retrieval impairment. This impairment was transient, and memory returned to control levels at later times. In agreement with previous studies, we found that systemic administration of propranolol disrupts the reconsolidation of Pavlovian FC and that its injection following a retrieval elicited by cue exposure also interferes with the reconsolidation of contextual FC. Hence, propranolol disrupts the reconsolidation of Pavlovian FC, but has no effect on the reconsolidation of IA. The results indicate that the efficacy of systemic administration of propranolol in disrupting the reconsolidation of fear memories is limited.

Delayed Increase of Brain Noradrenaline After Acute Footshock Stress in Rats

Several lines of evidence strongly suggest that accumulation of noradrenaline (NA) in the brain may underlie the hyperarousal symptoms experienced in post-traumatic stress disorder. In animal experiments, however, the effect of stress on NA content appears complex; acute stress reduces the level, while chronic stress tends to increase it. To explain this discrepancy, it is necessary to observe the long-term effects of acute stress on NA metabolism in the brain. In this study, rats were exposed to intermittent intense footshock stress for 1 h, and the brain NA content was measured for 7 days after the stress stimulus. Hypothalamic NA content was immediately reduced and recovered within 24 h. However, a significant NA increase was observed 7 days after the footshock. In the cerebral cortex and hippocampus, an increase in NA content was observed 1 day after the stress and lasted for at least 7 days. The fact that the content of 3-methoxy-4-hydroxyphenylglycol, a major NA metabolite, only transiently increased in all these regions possibly reflects NA release. These results indicate that increase in the brain NA content can be induced by acute stress, though its emergence is delayed. Importantly, this suggests that both acute and chronic stress may lead to NA accumulation under the same mechanism.

Endocannabinoids mediate acute fear adaptation via glutamatergic neurons independently of corticotropin‐releasing hormone signaling

Recent evidence showed that the endocannabinoid system plays an important role in the behavioral adaptation of stress and fear responses. In this study, we chose a behavioral paradigm that includes criteria of both fear and stress responses to assess whether the involvement of endocannabinoids in these two processes rely on common mechanisms. To this end, we delivered a footshock and measured the fear response to a subsequently presented novel tone stimulus. First, we exposed different groups of cannabinoid receptor type 1 (CB(1))-deficient mice (CB(1) (-/-)) and their wild-type littermates (CB(1) (+/+)) to footshocks of different intensities. Only application of an intense footshock resulted in a sustained fear response to the tone in CB(1) (-/-). Using the intense protocol, we next investigated whether endocannabinoids mediate their effects via an interplay with corticotropin-releasing hormone (CRH) signaling. Pharmacological blockade of CB(1) receptors by rimonabant in mice deficient for the CRH receptor type 1 (CRHR1(-/-)) or type 2 (CRHR2(-/-)), and in respective wild-type littermates, resulted in a sustained fear response in all genotypes. This suggests that CRH is not involved in the fear-alleviating effects of CB(1). As CRHR1(-/-) are known to be severely impaired in stress-induced corticosterone secretion, our observation also implicates that corticosterone is dispensable for CB(1)-mediated acute fear adaptation. Instead, conditional mutants with a specific deletion of CB(1) in principal neurons of the forebrain (CaMK-CB(1) (-/-)), or in cortical glutamatergic neurons (Glu-CB(1) (-/-)), showed a similar phenotype as CB(1) (-/-), thus indicating that endocannabinoid-controlled glutamatergic transmission plays an essential role in acute fear adaptation.

Prenatal stress exacerbates the impact of an aversive procedure on the corticosterone response to stress in female rats

Post-traumatic stress disorder (PTSD) is associated with marked alterations in hypothalamic-pituitary adrenal (HPA) function. In rats, prenatal restraint stress (stress applied to pregnant mothers, PRS) is known to impact behavioral and neuroendocrine sensitivity to several kinds of mild stress in adulthood. We have recently shown that PRS also modifies behavioral responses after exposure to an intense footshock in a potential animal model of PTSD. The aim of the present study was to evaluate the long-term effects of an aversive procedure (footshock followed by 3 weekly situational reminders) on the corticosterone response to a novel stress (restraint stress, 140 days after the footshock) in adult female PRS rats. Our data extend previous results showing that PRS leads to a long-lasting increase in plasma corticosterone after restraint stress in adult male rats. Moreover, we demonstrate that 140 days after the intense footshock, female PRS rats have lower corticosterone levels 60 min after restraint stress, suggesting an increase in the negative feedback of the HPA axis. These results indicate that early stress may favor long-lasting modifications of the HPA axis subsequent to exposure to an intense stress in adulthood.

Post-training intra-hippocampal infusions of corticosterone promote, preserve or disrupt conditioned fear responses depending on the stressful situation

Event Abstract Back to Event Post-training intra-hippocampal infusions of corticosterone promote, preserve or disrupt conditioned fear responses depending on the stressful situation Nadia Kaouane1*, Yves Porte1, Ludovic Calandreau1, Pier Vincenzo-Piazza2 and Aline Desmedt1 1 CNRS UMR 5228, Centre de Neurosciences Integratives et Cognitives, France 2 Neurocentre Magendie, INSERM U862, France Extensive evidence indicates that systemic or intra-hippocampal administration of adrenocortical hormone corticosterone (CORT) facilitates the consolidation of emotional experience. However, high CORT release, as shown under extreme stressful situation, can induce memory deficits. This suggests that beyond a certain level of stress, the influence of CORT on memory consolidation may switch from a facilitating to a deleterious effect, leading to maladaptive emotional responses with regard to the learning experience. Nevertheless, the boundary conditions under which such a switch can be observed are still elusive. To specify these conditions, we used in mice two fear conditioning procedures known to result in a preferential conditioned fear response to either a discrete tone (i. e. tone-shock pairing) or contextual cues (i. e. tone-shock unpairing). First, we show that increasing the intensity of footshock from a very low (0.3 mA) to a relatively high (1 mA) level gradually enhances conditioned fear responses in an adaptive manner, i.e. as a function of the tone-shock contingency. However, beyond the critical footshock intensity of 1 mA, fear responses become maladaptive, i.e. independent on the conditioning procedure. Second, depending on the footshock intensity (0.3 mA, 0.5 mA, 0.7 mA) and the dose used (0, 1, 10, 20, 50, 100 ng per side), post-training intra-hippocampal infusions of CORT promote, preserve or disrupt conditioned fear responses. Finally, we also assessed whether the CORT-dependent switch from adaptive to maladaptive fear responses was associated with changes in c-fos induction-related recruitment of the prefronto-hippocampo-amygdalar circuit. Altogether, these results throw light on the boundary conditions of CORT-dependent normal versus pathological fear learning. Conference: 41st European Brain and Behaviour Society Meeting, Rhodes Island, Greece, 13 Sep - 18 Sep, 2009. Presentation Type: Poster Presentation Topic: Poster presentations Citation: Kaouane N, Porte Y, Calandreau L, Vincenzo-Piazza P and Desmedt A (2009). Post-training intra-hippocampal infusions of corticosterone promote, preserve or disrupt conditioned fear responses depending on the stressful situation. Conference Abstract: 41st European Brain and Behaviour Society Meeting. doi: 10.3389/conf.neuro.08.2009.09.186 Copyright: The abstracts in this collection have not been subject to any Frontiers peer review or checks, and are not endorsed by Frontiers. They are made available through the Frontiers publishing platform as a service to conference organizers and presenters. The copyright in the individual abstracts is owned by the author of each abstract or his/her employer unless otherwise stated. Each abstract, as well as the collection of abstracts, are published under a Creative Commons CC-BY 4.0 (attribution) licence (https://creativecommons.org/licenses/by/4.0/) and may thus be reproduced, translated, adapted and be the subject of derivative works provided the authors and Frontiers are attributed. For Frontiers’ terms and conditions please see https://www.frontiersin.org/legal/terms-and-conditions. Received: 10 Jun 2009; Published Online: 10 Jun 2009. * Correspondence: Nadia Kaouane, CNRS UMR 5228, Centre de Neurosciences Integratives et Cognitives, Talence, France, n.kaouane@cnic.u-bordeaux1.fr Login Required This action requires you to be registered with Frontiers and logged in. To register or login click here. Abstract Info Abstract The Authors in Frontiers Nadia Kaouane Yves Porte Ludovic Calandreau Pier Vincenzo-Piazza Aline Desmedt Google Nadia Kaouane Yves Porte Ludovic Calandreau Pier Vincenzo-Piazza Aline Desmedt Google Scholar Nadia Kaouane Yves Porte Ludovic Calandreau Pier Vincenzo-Piazza Aline Desmedt PubMed Nadia Kaouane Yves Porte Ludovic Calandreau Pier Vincenzo-Piazza Aline Desmedt Related Article in Frontiers Google Scholar PubMed Abstract Close Back to top Javascript is disabled. Please enable Javascript in your browser settings in order to see all the content on this page.

Associative structure of fear memory after basolateral amygdala lesions in rats.

The authors have recently demonstrated that rats with basolateral amygdala (BLA) lesions acquire Pavlovian fear conditioning after overtraining. However, it is not known whether the associative basis of Pavlovian fear memory acquired by rats with BLA lesions is similar to that of intact rats. Associations are typically formed between the conditional (CS) and unconditional (US) stimuli (stimulus-stimulus; S-S), although it is possible for stimuli to enter into association with the responses they produce (stimulus-response; S-R). Indeed, the central nucleus of the amygdala, which is essential for fear conditioning in rats with BLA lesions, may mediate S-R associations in some Pavlovian tasks. The authors therefore used a postconditioning US inflation procedure (i.e., exposure to intense footshock USs) to assess the contribution of S-S associations to fear conditioning after overtraining in rats with BLA lesions. In Experiment 1, intact rats that were overtrained and later inflated displayed elevated freezing levels when tested, indicating that S-S associations contribute to overtrained fear memories. Interestingly, neither neurotoxic BLA lesions nor temporary inactivation of the BLA during overtraining prevented the inflation effect (Experiment 2 and 3, respectively). These results reveal that S-S associations support Pavlovian fear memories after overtraining in both intact rats and rats with BLA lesions, and imply that the central nucleus of the amygdala encodes CS-US associations during fear conditioning.

Enhanced inhibitory avoidance learning prevents the long-term memory-impairing effects of cycloheximide, a protein synthesis inhibitor

Interference with activity of numerous cerebral structures produces memory deficiencies; in many instances, however, when animals are over-trained such interference becomes innocuous. Systemic administration of protein synthesis inhibitors impairs long-term retention; this effect has been interpreted to mean that protein synthesis is required for memory consolidation, though little is known about the effect of protein synthesis inhibitors on memory of enhanced learning in the rat. To further analyze the protective effect of enhanced learning against amnesic treatments, groups of Wistar rats were trained in a one-trial step-through inhibitory avoidance task, using different intensities of foot-shock during training. Cycloheximide (CXM; 2.8 mg/kg), an inhibitor of protein synthesis, was injected either 30 min before training or immediately after training. Twenty-four hours after training retention latencies were recorded. Our data showed that both pre- and post-training administration of CXM produced amnesia in those groups that had been trained with relatively low foot-shock intensities, but no impairment in retention was observed when relatively high intensities of foot-shock were administered. These and similar results lead us to conclude that protein synthesis inhibitors may interfere with memory consolidation, but their effect disappears when animals are submitted to an enhanced learning experience, calling into question the idea that protein synthesis is required for memory consolidation.

Dorsal hippocampus involvement in delay fear conditioning depends upon the strength of the tone‐footshock association

The hippocampus is important for the formation of spatial, contextual, and episodic memories. For instance, lesions of the dorsal hippocampus (DH) produce demonstrable deficits in contextual fear conditioning. By contrast, it is generally agreed that the DH is not important for conditioning to a discrete cue (such as a tone or light) that is paired with footshock in a temporally contiguous fashion (delay conditioning). There are, however, some reports of hippocampus involvement in delay conditioning. The present series of experiments was designed to assess the conditions under which the hippocampus-dependent component of delay fear conditioning performance may be revealed. Here, we manipulated the number of conditioning trials and the intensity of the footshock in order to vary the strength of conditioning. The results indicate that the DH contributes to freezing performance to a delay conditioned tone when the conditioning parameters are relatively weak (few trials or low footshock intensity), but not when strong parameters are used. The results are discussed in terms of two parallel memory systems: a direct tone-footshock association that is independent of the hippocampus and a hippocampus-dependent memory for the conditioning session.

Acquisition and retention of enhanced active avoidance are unaffected by interference with serotonergic activity

Pre-training administration of p-chloroamphetamine (PCA) produces reliable deficits of avoidance learning. When animals are trained in inhibitory avoidance with relatively high foot-shock intensities, other amnesic treatments have no effect. The present experiment was conducted to determine if this protective effect of high foot shock is also observed after administration of PCA (10 mg/kg, i.p., injected 7 days before training; this dose produces a lesion of central serotonin neurons). Rats were trained in active avoidance (a single 20-trial session), administering shocks of 0.6, 0.8, 1.0, or 1.4 mA to independent groups of rats. When compared to saline-injected groups trained with the same intensities, PCA produced a significant learning deficit in the low foot-shock groups, but not in the high foot-shock animals. These results indicate that the dose of PCA administered, which is known to deplete cerebral serotonin, does not interfere with acquisition and retention of enhanced active avoidance training.

Targeted deletion of the GABRA2 gene encoding α2-subunits of GABA A receptors facilitates performance of a conditioned emotional response, and abolishes anxiolytic effects of benzodiazepines and barbiturates

Mice with point-mutated α2 GABA A receptor subunits (rendering them diazepam insensitive) are resistant to the anxiolytic-like effects of benzodiazepines (BZs) in the conditioned emotional response (CER) test, but show normal anxiolytic effects of a barbiturate. We investigated the consequence of deleting the α2-subunit on acquisition of the CER with increasing intensity of footshock, and on the anxiolytic efficacy of a benzodiazepine, diazepam, and a barbiturate, pentobarbital. α2 knockout (KO) and wildtype (WT) mice were trained in a conditioned emotional response (CER) task, in which lever pressing for food on a variable interval (VI) schedule was suppressed during the presentation of a compound light/tone conditioned stimulus (CS+) that predicted footshock. The ability of diazepam and of pentobarbital to reduce suppression during the CS+ was interpreted as an anxiolytic response. There were no differences between the genotypes in shock sensitivity, as assessed by their flinch responses to increasing levels of shock. However, α2 KO mice showed a greater suppression of lever pressing than WT littermates in the presence of a compound cue signalling footshock. Diazepam (0, 0.5, 1 and 2 mg/kg) induced a dose-dependent anxiolytic-like effect in WT mice but no such effect was seen in KO mice. Similarly, although pentobarbital (20 mg/kg) reduced the ability of the CS+ to reduce lever pressing rates in WT mice, this effect was not seen in the KO. These findings suggest that α2-containing GABA A receptors mediate the anxiolytic effects of barbiturates, as well as benzodiazepines, and that they may be involved in neuronal circuits underlying conditioned anxiety.

Opioid receptors mediate direct predictive fear learning: Evidence from one-trial blocking

Pavlovian fear learning depends on predictive error, so that fear learning occurs when the actual outcome of a conditioning trial exceeds the expected outcome. Previous research has shown that opioid receptors, including mu-opioid receptors in the ventrolateral quadrant of the midbrain periaqueductal gray (vlPAG), mediate such predictive fear learning. Four experiments reported here used a within-subject one-trial blocking design to study whether opioid receptors mediate a direct or indirect action of predictive error on Pavlovian association formation. In Stage I, rats were trained to fear conditioned stimulus (CS) A by pairing it with shock. In Stage II, CSA and CSB were co-presented once and co-terminated with shock. Two novel stimuli, CSC and CSD, were also co-presented once and co-terminated with shock in Stage II. The results showed one-trial blocking of fear learning (Experiment 1) as well as one-trial unblocking of fear learning when Stage II training employed a higher intensity footshock than was used in Stage I (Experiment 2). Systemic administrations of the opioid receptor antagonist naloxone (Experiment 3) or intra-vlPAG administrations of the selective mu-opioid receptor antagonist CTAP (Experiment 4) prior to Stage II training prevented one-trial blocking. These results show that opioid receptors mediate the direct actions of predictive error on Pavlovian association formation.