An immediate-shock freezing deficit with discrete cues: a possible role for unconditioned stimulus processing mechanisms.

Five experiments with C57BL/6 mice (Mus musculus) investigated whether failures in shock processing might contribute to deficits in freezing that occur after an animal receives a shock immediately on exposure to a conditioning context. Experiment 1 found that more contextual freezing resulted from delayed shocks than from immediate shocks across 4 shock intensities. Experiment 2 extended the immediate-shock freezing deficit to discrete stimuli. Experiment 3 found that preexposure to the to-be-conditioned cue did not facilitate immediate cued conditioning. Experiment 4 found that context preexposure enhanced context-evoked fear after an immediate shock. Experiment 5 found that context preexposure also enhanced immediate cued conditioning. These findings are problematic for current theories of the immediate-shock freezing deficit that focus exclusively on processing of the conditioned stimulus, and they suggest that failures in shock processing may contribute to the deficit.     

Exposure to a stressor produces a long lasting enhancement of fear learning in rats

In contextual fear conditioning, footshock is given in a context, and re-exposure to this context elicits the conditional defensive response of freezing, a reliable behavioral index of conditional fear. Normally, the amount of contextual freezing is directly proportional to the number of shocks an animal receives in the context. However, pre-exposure to a stressor can produce an enhancement in conditional freezing. Pre-exposure to repeated footshock in one context produces an enhancement of conditional freezing to cues associated with a single shock in a second distinct context. This model of stress-enhanced fear learning (SEFL) can be utilized to study how stress affects learning of future aversive events. The experiments in this paper characterize the magnitude and longevity of SEFL. In the first experiment, the number of footshocks given during the pre-exposure session was varied and conditional fear to the single shock was assessed. Pre-exposure to 1 shock did not produce an enhancement in fear learning in the second context, but pre-exposure to 4 or 15 shocks did. The time-course of the enhancement was examined in the next two experiments. These experiments show that SEFL persists for at least 3 months.     

The role of injection cues in the associative control of the US pre-exposure effect in flavour aversion learning

Two experiments, using rats as subjects, examined the role of contextual cues in producing the unconditioned stimulus (US) pre-exposure effect in conditioned taste aversion. Experiment 1 showed a significant US pre-exposure effect, when the pre-exposure was conducted in a familiar context, and that a change of context between the pre-exposure and conditioning phases did not attenuate this effect. Experiment 2 demonstrated that extinction of injection-related cues after the pre-exposure stage attenuated the US pre-exposure effect, when the pre-exposure was conducted in either a familiar or a novel context. Taken together, these results support the associative explanation of the US pre-exposure effect in terms of blocking, incorporating a role for injection-related cues in the context blocking analysis of the US pre-exposure effect.     

Effects of environmental enrichment on rate of contextual processing and discriminative ability in adult rats

The effect of environmental enrichment on conditioned freezing to contextual cues in adult Sprague-Dawley rats was examined. The freezing of both enriched-and standard-reared rats increased with the time spent in the chamber prior to shock. Both groups of rats showed equally low levels of contextual conditioning following a preshock period of 4 s and equally high levels following a 120-s preshock period. However, following a preshock period of 16 s enriched rats displayed more contextual conditioning than standard rats. That is, enriched rats appeared to process contextual information faster than their standard-reared counterparts. Enriched- reared rats also showed a greater ability to discriminate between the conditioning context and a similar but distinctive context. Hence, in addition to forming a representation of the context in memory more rapidly than standard-reared rats, enriched-reared rats also appear to form a more complex representation.     

Dissociated roles for the lateral and medial septum in elemental and contextual fear conditioning

Extensive evidence indicates that the septum plays a predominant role in fear learning, yet the direction of this control is still a matter of debate. Increasing data suggest that the medial (MS) and lateral septum (LS) would be differentially required in fear conditioning depending on whether a discrete conditional stimulus (CS) predicts, or not, the occurrence of an aversive unconditional stimulus (US). Here, using a tone CS-US pairing (predictive discrete CS, context in background) or unpairing (context in foreground) conditioning procedure, we show, in mice, that pretraining inactivation of the LS totally disrupted tone fear conditioning, which, otherwise, was spared by inactivation of the MS. Inactivating the LS also reduced foreground contextual fear conditioning, while sparing the higher level of conditioned freezing to the foreground (CS-US unpairing) than to the background context (CS-US pairing). In contrast, inactivation of the MS totally abolished this training-dependent level of contextual freezing. Interestingly, inactivation of the MS enhanced background contextual conditioning under the pairing condition, whereas it reduced foreground contextual conditioning under the unpairing condition. Hence, the present findings reveal a functional dissociation between the LS and the MS in Pavlovian fear conditioning depending on the predictive value of the discrete CS. While the requirement of the LS is crucial for the appropriate processing of the tone CS-US association, the MS is crucial for an appropriate processing of contextual cues as foreground or background information.     

Nicotine produces a within-subject enhancement of contextual fear conditioning in C57BL/6 mice independent of sex

Nicotine enhances learning including contextual fear conditioning. The present study extends previous work on nicotine and conditioned fear to examine the nature of nicotine’s enhancement of contextual fear conditioning and sex differences in contextual fear conditioning in C57BL/6 mice using a within-subjects design. Mice were trained by pairing of an auditory stimulus of 80 dB, 6 cps train of broad-band clicks conditioned stimulus (CS) with a 2 sec., 0.35 mA shock unconditioned stimulus (US). Twenty-four hours later mice were tested for freezing in the original context, and one hour later mice were retested in the same context. A 0.5 mg/kg dose of nicotine was given either for three conditions: (1) before training, testing, and retesting; (2) before training and retesting; and (3) before retesting only. The use of a within-subjects design allowed for testing if nicotine would produce state-dependent deficits in contextual fear conditioning. Nicotine did enhance contextual fear conditioning in the groups that received nicotine for both training and testing. Nicotine, however, did not alter freezing when given on training but not testing or testing but not training. No sex differences, however, existed for conditioning or for nicotine’s effects on conditioning. These results suggest that nicotine enhanced acquisition and retrieval processes but did not produc state-dependent deficits when administered just for training or just for testing.     

Attenuation of latent inhibition by post-acquisition reminder

Using lick suppression by water-deprived rats as an associative index, white noise-footshock pairings resulted in less manifest conditioning when repeated non-reinforced presentations of the white noise preceded conditioning than when no stimulus pre-exposure was given, i.e., latent inhibition was observed. However, the latent inhibition deficit was reduced in animals who received as a reminder treatment shock-alone presentations in another context during the retention interval. Animals conditioned without prior stimulus pre-exposure and those exposed to the white noise and shock unpaired during the conditioning phase of the study showed no change in lick suppression as a result of the reminder treatment. These results suggest that the behavioural deficit produced by non-reinforced pre-exposure to the to-be-conditioned stimulus arises at least in part from a reversible retrieval failure rather than a lack of acquisition.     

Latent inhibition, context specificity, and context familiarity

In two appetitive licking experiments, thirsty rats were pre-exposed to two stimuli prior to conditioning sessions in which one or both of these stimuli were paired with water. Consistent with other results, latent inhibition was disrupted when conditioning took place in a context different from that in which stimulus pre-exposure had occurred. But in animals given prior exposure to the context of stimulus pre-exposure before the start of stimulus pre-exposure, substantial and equivalent latent inhibition was evident whether or not there was a change of context between stimulus pre-exposure and conditioning. These results are discussed in terms of current theories of latent inhibition.     

Interactive learning in the paradise fish (Macropodus opercularis): an ethological interpretation of the second-order conditioning paradigm

This study was aimed at the examination of ‘mental construction’ in paradise fish by interactive learning, which is suggested as an alternative hypothesis for backward or second-order conditioning. Avoidance of goldfish was established in paradise fish by presenting a harmless goldfish (a novel stimulus) after an aversive event (mild electric shocks) in the dark compartment of a shuttle tank. It was found that this avoidance depended on context pre-exposure. Experiment 1 was designed to study the effect of mild shocks on shuttling activity in a familiar context. Experiment 2 aimed at establishing fear-conditioning to the goldfish in a higher-order conditioning paradigm. In the course of training, unpaired stimuli were presented in the dark compartment of the shuttle tank in such a manner that the presentation of mild shocks (20 mA) preceded the encounter with a harmless fish (goldfish). Experiment 3 demonstrated the role of context pre-exposure in interactive learning. Results indicate that (1) while 60 mA shocks resulted in avoidance of the dark compartment, the 20 mA mild shocks affected exploratory behaviour; (2) after pre-exposure to the training environment, paradise fish avoided the dark compartment containing goldfish provided that subjects had previously encountered mild, explorative shocks there; (3) this conditioned fear, a ‘mental construction’ of a potential danger, was proved to be transferable to another context and was consequently aimed specifically at the goldfish, a living being, rather than the training context; (4) the pre-exposure to the shuttle tank was an important part of this training procedure, that is, only subjects habituated to the shuttle tank avoided the goldfish. Results are discussed in the framework of the Interactive Learning Hypothesis, which has been developed as an ethological approach to a higher-order conditioning paradigm. Received: 15 January 1999 / Accepted after revision: 18 August 1999     

Roles of hippocampal GABA(A) and muscarinic receptors in consolidation of context memory and context-shock association in contextual fear conditioning: a double dissociation study

Contextual fear conditioning involves forming a context representation and associating it to a shock, both of which involved the dorsal hippocampus (DH) according to our recent findings. This study tested further whether the two processes may rely on different neurotransmitter systems in the DH. Male Wistar rats with cannula implanted into the DH were subjected to a two-phase training paradigm of contextual fear conditioning to separate context learning from context-shock association in two consecutive days. Immediately after each training phase, different groups of rats received bilateral intra-DH infusion of the GABA(A) agonist muscimol, 5HT(1A) agonist 8-OH-DPAT, NMDA antagonist APV or muscarinic antagonist scopolamine at various doses. On the third day, freezing behavior was tested in the conditioning context. Results showed that intra-DH infusion of muscimol impaired conditioned freezing only if it was given after context learning. In contrast, scopolamine impaired conditioned freezing only if it was given after context-shock training. Posttraining infusion of 8-OH-DPAT or APV had no effect on conditioned freezing when the drug was given at either phase. These results showed double dissociation for the hippocampal GABAergic and cholinergic systems in memory consolidation of contextual fear conditioning: forming context memory required deactivation of the GABA(A) receptors, while forming context-shock memory involved activation of the muscarinic receptors.     

Nicotine Produces a Within-Subject Enhancement of Contextual Fear Conditioning in C57BL/6 Mice Independent of Sex

Nicotine enhances learning including contextual fear conditioning. The present study extends previous work on nicotine and conditioned fear to examine the nature of nicotine's enhancement of contextual fear conditioning and sex differences in contextual fear conditioning in C57BL/6 mice using a within-subjects design. Mice were trained by pairing of an auditory stimulus of 80 dB, 6 cps train of broad-band clicks conditioned stimulus (CS) with a 2 sec., 0.35 mA shock unconditioned stimulus (US). Twenty-four hours later mice were tested for freezing in the original context, and one hour later mice were retested in the same context. A 0.5 mg/kg dose of nicotine was given either for three conditions: (1) before training, testing, and retesting; (2) before training and retesting; and (3) before retesting only. The use of a within-subjects design allowed for testing if nicotine would produce state-dependent deficits in contextual fear conditioning. Nicotine did enhance contextual fear conditioning in the groups that received nicotine for both training and testing. Nicotine, however, did not alter freezing when given on training but not testing or testing but not training. No sex differences, however, existed for conditioning or for nicotine's effects on conditioning. These results suggest that nicotine enhanced acquisition and retrieval processes but did not produce state-dependent deficits when administered just for training or just for testing.     

Contextual control over conditioned responding in a latent inhibition paradigm

We used 1-, 2-, and 3-context designs to study the control exerted by contexts over freezing in rats exposed to a conditioned stimulus (CS) in advance of its pairing with a shock unconditioned stimulus. The latent inhibition observed when preexposure, conditioning, and testing occurred in the same context was attenuated if preexposure occurred in a different context to conditioning and testing. Latent inhibition (i.e., attenuated performance) was restored in a CS-specific manner if preexposure and testing occurred in the same context and conditioning in a different one. Latent inhibition was also reduced by a long retention interval but remained specific for a particular context-CS relation. Finally, CS preexposure resulted in contextual control over the expression of excitatory conditioned performance. The results are discussed in terms of memory, associative, and associative-performance models of CS-preexposure effects.     

Unpaired shocks during extinction weaken the contextual renewal of a conditioned discrimination

Extinction is generally more fragile than conditioning, as illustrated by the contextual renewal effect. The traditional extinction procedure entails isolated presentations of the conditioned stimulus. Extinction may be boosted by adding isolated presentations of the unconditioned stimulus, as this should augment breaking the contingency between the two stimuli. In a human conditioning experiment with on-line expectancy ratings and electrodermal responding as dependent variables, 32 participants were differentially conditioned to two neutral figures using electric shock. After a change of context, one group received normal extinction treatment whereas another group received explicitly unpaired presentations of the figures and shock. At test, the two figures were presented in the original context again. For both measures, only the group that received normal extinction showed renewal of the conditioned discrimination. These results suggest that unpaired shocks during extinction strengthen the extinction learning.     

Effects of multisensory environmental stimulation on contextual conditioning in the developing rat

The effect of increased exposure to multisensory stimulation during development on conditioned freezing to contextual cues in preweanling Sprague-Dawley rats was examined. Rats given increased environmental stimulation exhibited long-term contextual conditioning at a younger age than rats that did not receive such stimulation when there was either low or moderate levels of conditioning (Experiments 1 and 2). These differences in contextual conditioning were not a result of the stimulated rats reacting differently to shock (Experiment 4) or merely freezing more than the nonstimulated rats in all situations (Experiment 3). The role of the glucocorticoid system in the enhanced contextual learning of stimulated preweanling rats and the advantages of the contextual conditioning procedure for studying the effects of environmental stimulation are discussed.     

Temporally graded,context-specific retrograde amnesia and its alleviation by context preexposure: effects of postconditioning exposures to morphine in the rat

Five experiments studied retrograde impairments in Pavlovian fear conditioning following prolonged exposure to the opioid receptor agonist morphine. Injections of morphine commencing 1-7 days but not 14 days after conditioning produced amnesia for that conditioning episode. This amnesia was (a) selective such that morphine impaired freezing to the conditioning context but not to the auditory conditioned stimulus, (b) independent of the interval between the last injection of morphine and test, and (c) accompanied by a failure of contextual discrimination. Context preexposure protected context conditioning and discrimination from the amnestic effects of morphine. These results show that retrograde deficits in contextual fear conditioning are mediated by failures to consolidate a contextual representation.     

Effects of multiple exposures to D-cycloserine on extinction of conditioned fear in rats

Previous research has shown that an acute, post-training injection of D-cycloserine (DCS) facilitates extinction of conditioned fear in rats; however, the effects of multiple exposures to DCS in this situation are not known. In Experiment 1, rats were conditioned (light-shock pairings) and 24 h later given six extinction (light-alone) trials followed by an injection of DCS (15 mg/kg) or saline. The next day, all rats were tested for light-elicited freezing. In Experiment 2, the effect of DCS on extinction was tested in the same manner, except that rats were pre-exposed to DCS (0, 1, or 5 injections) just prior to conditioning. In Experiment 3, rats received five pre-exposures of DCS but conditioning occurred either 2 or 28 days after the last pre-exposure. The results showed that DCS facilitated extinction of conditioned freezing to the light CS when no drug pre-exposure had occurred, but pre-exposure to DCS just prior to conditioning disrupted the facilitation of extinction effect. When 28 days were interposed between pre-exposure and conditioning, the facilitatory effects of DCS on extinction were restored. These findings suggest that DCS has significant clinical value but that behavioral desensitization may occur with multiple exposures; however, desensitization is not permanent and is reduced by the passage of time.     

Infusion of lidocaine into the dorsal hippocampus before or after the shock training phase impaired conditioned freezing in a two-phase training task of contextual fear conditioning

Learning in a contextual fear conditioning task involves forming a context representation and associating it with a shock. The dorsal hippocampus (DH) is implicated in representing the context, but whether it also has a role in associating the context and shock is unclear. To address this issue, male Wistar rats were trained on the task by a two-phase training paradigm, in which rats learned the context representation on day 1 and then reactivated it to associate with the shock on day 2; conditioned freezing was tested on day 3. Lidocaine was infused into the DH at various times in each of the two training sessions. Results showed that intra-DH infusion of lidocaine shortly before or after the context training session on day 1 impaired conditioned freezing, attesting to the DH involvement in context representation. Intra-DH infusion of lidocaine shortly before or after the shock training session on day 2 also impaired conditioned freezing. This deficit was reproduced by infusing lidocaine or APV (alpha-amino-5-phosphonovaleric acid) into the DH after activation of the context memory but before shock administration. The deficit was not due to drug-induced state-dependency, decreased shock sensitivity or reconsolidation failure of the contextual memory. These results suggest that in contextual fear conditioning integrity of the DH is required for memory processing of not only context representation but also context-shock association.     

A search for context-stimulus associations in latent inhibition

Six experiments investigated the effects of pre-exposure to a tone on the subsequent acquisition of conditioned suppression by rats. In Experiments 1-3 the response suppressed was drinking; in Experiments 4-6 it was food-rewarded lever pressing. Repeated exposure to the tone resulted in latent inhibition, i.e., a retardation in the acquisition of suppression. The size of the latent inhibition effect was reduced when a different context was used for conditioning from that used for pre-exposure (Experiment 2). When the context remained the same throughout, a phase of exposure to the context alone, interposed between pre-exposure and conditioning, had no influence on the size of the latent inhibition effect ultimately observed. This last result casts doubt upon Wagner's (1976, 1979) theory of the role of contextual factors in latent inhibition, and alternative accounts are considered.     

Conditioned and unconditioned fear organized in the periaqueductal gray are differentially sensitive to injections of muscimol into amygdaloid nuclei

The lateral and basolateral nuclei of the amygdala (LaA and BLA, respectively) serve as a filter for unconditioned and conditioned aversive information that ascends to higher structures from the brainstem, whereas the central nucleus of the amygdala (CeA) is considered to be the main output for the defense reaction. It has been shown that the dorsal periaqueductal gray (dPAG) is activated by threatening stimuli and has important functional links with the amygdala through two-way anatomical connections. In this work, we examined the influence of chemical inactivation of these nuclei of amygdala on the freezing and escape responses induced by electrical stimulation through electrodes implanted in the dPAG of Wistar rats. Each rat also bore a cannula implanted in the LaA, BLA or CeA for injections of muscimol (0.5 microg/0.5 microL) or its vehicle. The duration of freezing behavior that outlasts electrical stimulation of the dPAG was also measured. On the following day, these animals were submitted to a contextual fear-conditioning using foot shocks as unconditioned stimulus. Conditioned freezing to contextual cues previously associated with foot shocks was also inhibited by injections of muscimol into these amygdaloid nuclei. The contextual conditioned freezing behavior is generated in the neural circuits of conditioned fear in the amygdala. The data obtained also show that injections of muscimol into the three amygdaloid nuclei did not change the aversive threshold of freezing, but disrupted the dPAG post-stimulation freezing. Previous findings that the latter freezing results directly from dPAG stimulation and that it is not sensitive to a context shift suggest that it is unconditioned in nature. Thus, the amygdala can affect some, but not all, aspects of unconditioned freezing. Post-stimulation freezing may reflect the process of transferring aversive information from dPAG to higher brain structures.     

Cross-modal transfer of conditioned suppression in rats: Effects of US intensity and extinction of the initial conditioning task

In conditioned suppression of water licking behavior by rats, we obtained data indicating general transfer of fear conditioning. A series of experiments resulted in two major findings. First, pairing of a neutral stimulus with a shock in the initial conditioning task facilitated acquisition of subsequent fear conditioning to another neutral stimulus, if the conditioned fear of the initial task was extinguished prior to the second task and if equally strong shocks were employed in both conditioning tasks. Second, omission of the extinction treatment or employment of weaker shocks in the initial task resulted in retardation, rather than facilitation, of the second conditioning task. An application to human clinical settings is discussed.