Timing of fear expression in trace and delay conditioning measured by fear-potentiated startle in rats

In two experiments, the time course of the expression of fear in trace (hippocampus-dependent) versus delay (hippocampus-independent) conditioning was characterized with a high degree of temporal specificity using fear-potentiated startle. In experiment 1, groups of rats were given delay fear conditioning or trace fear conditioning with a 3- or 12-sec trace interval between conditioned stimulus (CS) offset and unconditioned stimulus (US) onset. During test, the delay group showed fear-potentiated startle in the presence of the CS but not after its offset, whereas the trace groups showed fear-potentiated startle both during the CS and after its offset. Experiment 2 compared the time course of fear expression after trace conditioning with the time course in two delay conditioning groups: one matched to the trace conditioning group with respect to CS duration, and the other with respect to ISI. In all groups, fear was expressed until the scheduled occurrence of the US and returned to baseline rapidly thereafter. Thus, in both trace and delay fear conditioning, ISI is a critical determinant of the time course of fear expression. These results are informative as to the possible role of neural structures, such as the hippocampus, in memory processes related to temporal information.     

Effects of Early Hippocampal Lesions on Trace, Delay, and Long-Delay Eyeblink Conditioning in Developing Rats

The effects of bilateral hippocampal aspiration lesions on later acquisition of eyeblink conditioning were examined in developing Long-Evans rat pups. Lesions on postnatal day (PND) 10 were followed by evaluation of trace eyeblink conditioning (Experiment 1) and delay eyeblink conditioning (Experiment 2) on PND 25. Pairings of a tone conditioned stimulus (CS) and periocular shock unconditioned stimulus (US, 100 ms) were presented in one of three conditioning paradigms: trace (380 ms CS, 500 ms trace interval, 880 ms interstimulus interval [ISI]), standard delay (380 ms CS, 280 ms ISI), or long delay (980 ms CS, 880 ms ISI). The results of two experiments indicated that hippocampal lesions impaired trace eyeblink conditioning more than either type of delay conditioning. In light of our previous work on the ontogeny of trace, delay, and long-delay eyeblink conditioning (Ivkovich, Paczkowski, & Stanton, 2000) showing that trace and long-delay eyeblink conditioning had similar ontogenetic profiles, the current data suggest that during ontogeny hippocampal maturation may be more important for the short-term memory component than for the long-ISI component of trace eyeblink conditioning. The late development of conditioning over long ISIs may depend on a separate process such as protracted development of cerebellar cortex.     

The role of local context in autoshaping

Three experiments used an autoshaping procedure with pigeons to examine the effects of nonreinforced, nontarget stimuli (ITI-fillers) during the intertrial interval on responding to a reinforced target CS. Experiment 1 replicated a previous demonstration that an ITI-filler that occupied a substantial portion of the ITI attenuated responding to the target CS relative to a group trained with a similar ITI but lacking the ITI-filler. Experiment 2 found that the superiority of responding typically found with a long ITI relative to a short ITI, that is, the trial-spacing effect, can be reversed by imposing a filler stimulus during the ITI in the former condition. Experiment 3, using a within-subject design, found that when one background stimulus condition occupied a majority of the interreinforcement interval, pairings of one target CS with reinforcement that were embedded within this background condition, that is, a long duration local context occurred for this CS, yielded better performance that a second, reinforced, target CS paired with reinforcement in the shorter duration background condition. These results confirmed predictions derived from a local context view of cycle time. Comparator theories of classical conditioning require incorporation of this notion into their conceptualization of effective cycle time in order to explain the present findings.     

Hippocampal and cerebellar single-unit activity during delay and trace eyeblink conditioning in the rat

In delay eyeblink conditioning, the CS overlaps with the US and only a brainstem-cerebellar circuit is necessary for learning. In trace eyeblink conditioning, the CS ends before the US is delivered and several forebrain structures, including the hippocampus, are required for learning, in addition to a brainstem-cerebellar circuit. The interstimulus interval (ISI) between CS onset and US onset is perhaps the most important factor in classical conditioning, but studies comparing delay and trace conditioning have typically not matched these procedures in this crucial factor, so it is often difficult to determine whether results are due to differences between delay and trace or to differences in ISI. In the current study, we employed a 580-ms CS-US interval for both delay and trace conditioning and compared hippocampal CA1 activity and cerebellar interpositus nucleus activity in order to determine whether a unique signature of trace conditioning exists in patterns of single-unit activity in either structure. Long-Evans rats were chronically implanted in either CA1 or interpositus with microwire electrodes and underwent either delay eyeblink conditioning, or trace eyeblink conditioning with a 300-ms trace period between CS offset and US onset. On trials with a CR in delay conditioning, CA1 pyramidal cells showed increases in activation (relative to a pre-CS baseline) during the CS-US period in sessions 1-4 that was attenuated by sessions 5-6. In contrast, on trials with a CR in trace conditioning, CA1 pyramidal cells did not show increases in activation during the CS-US period until sessions 5-6. In sessions 5-6, increases in activation were present only to the CS and not during the trace period. For rats with interpositus electrodes, activation of interpositus neurons on CR trials was present in all sessions in both delay and trace conditioning. However, activation was greater in trace compared to delay conditioning in the first half of the CS-US interval (during the trace CS) during early sessions of conditioning and, in later sessions of conditioning, activation was greater in the second half of the CS-US interval (during the trace interval). These results suggest that the pattern of hippocampal activation that differentiates trace from delay eyeblink conditioning is a slow buildup of activation to the CS, possibly representing encoding of CS duration or discrimination of the CS from the background context. Interpositus nucleus neurons show strong modeling of the eyeblink CR regardless of paradigm but show a changing pattern across conditioning that may be due to the necessary contributions of forebrain processing to trace conditioning.     

The Topography of Sexually Conditioned Behaviour: Effects of a Trace Interval

In a trace conditioning procedure, subjects were presented with a 30-sec conditioned stimulus (CS) followed by a 30-sec trace interval. Delayed conditioning consisted of a 60-sec CS presentation followed by an unconditioned stimulus (US). Although conditioning developed with both procedures, the topography of the conditioned response differed. Sexual conditioned approach was evident in all of the subjects during the presentation of the CS. Traceconditioned subjects moved away from the area where the CS had been presented during the trace interval but remained closer to the CS location than did an unpaired control. This reduction in the spatial specificity of the conditioned response was interpreted from a behaviour systems perspective. The trace interval presumably increased the perceived separation between the CS and the US and therefore elicited conditioned behaviour less specifically directed towards the CS.     

Both trace and delay conditioning of evaluative responses depend on contingency awareness

Whereas previous evaluative conditioning (EC) studies produced inconsistent results concerning the role of contingency knowledge, there are classical eye-blink conditioning studies suggesting that declarative processes are involved in trace conditioning but not in delay conditioning. In two EC experiments pairing neutral sounds (conditioned stimuli; CSs) with affective pictures (unconditioned stimuli; USs), we tested whether the type of conditioning procedure affects the relationship between awareness and EC. Auditory CSs and visual USs were either overlapping and co-terminating (Delay Conditioning Group), or they were separated by a short temporal gap (Trace Conditioning Group). In both groups, contingency awareness was manipulated by varying the duration at which the US was presented during conditioning. Furthermore, in addition to a post-conditioning measure of awareness, US detectability was measured concurrently during the learning phase in Experiment 2. US exposure duration affected both measures of awareness. The data of both experiments demonstrate that EC does not occur if the USs are presented at very brief durations, but reliable EC effects can be observed at longer US exposure durations. As there were no differences between trace and delay conditioning, the data suggest that contingency awareness plays a crucial role in trace and in delay conditioning of evaluative responses. These results challenge automatic association-formation accounts of EC, but they are in line with ‘single-process’ accounts assuming EC to be dependent on conscious declarative knowledge about CS–US contingencies.     

Latent inhibition: A trace conditioning phenomenon?

An informal model of latent inhibition (LI), the trace hypothesis, is described. This hypothesis holds that preexposure (PE) to the to-be-conditioned stimulus alters the salience (associability) of the CS such that only the onset and initial segments, but not the later segments, are capable of supporting conditioning when paired with a US. Thus, LI is effectively a trace conditioning phenomenon. Four experiments used rats and a one-trial fear-conditioning task to test predictions which stem from this hypothesis. Experiment 1 showed conditioning performance decreased as trace intervals increased between 0 and 12 s, thus demonstrating the sensitivity of this task to trace intervals proposed by the trace hypothesis to produce LI. Consistent with the trace hypothesis, LI was found to be an increasing function of the CS duration (Experiment 2), the number of PE trials (Experiment 3), and the US onset during a 15-s CS (Experiment 4). These data are somewhat problematic for several explanations of LI, but, in general, the trace hypothesis extends extant explanations by focusing on changes in salience within a given presentation of the CS.     

Variations in explicitly unpaired training are differentially effective in producing conditioned inhibition

To evaluate a contingency interpretation of conditioned inhibition (CI), rats were given “explicity unpaired” training in which the locus and duration of a CS within the inter-US (shock) interval were systematically manipulated for different groups. Summation and retardation tests in Experiment 1 indicated that stronger CI resulted from both a backward and a trace CS than from a midlocus CS of equal or greater duration. Complementing these findings, the same tests in Experiment 2 showed that, by comparison with novel-stimulus controls, CI developed to a trace CS but not to a mid-locus CS, nor to a trace CS that was accompanied by an immediate signal for the US. These findings argue against a contingency interpretation of CI and favor a contiguity interpretation stressing the short-term rehearsal of stimulus events. Such rehearsal of the US allows a backward CS, but not a mid-locus CS with an extended US-CS interval, to be discriminated as a signal for nonreinforcement, and thus to develop as a conditioned inhibitor. Similarly, excitatory conditioning to the memory trace of a CS allows the nominal trace CS to develop as a signal for nonreinforcement, and thus as a conditioned inhibitor, but not when its memory trace is overshadowed by another CS that immediately precedes the US. In short, the development of CI is facilitated when excitation is mediated by the memorial processing of either the US or a discrete CS for the US rather than by contextual cues.     

A comparison of latent inhibition and learned irrelevance pre-exposure effects in rabbit and human eyeblink conditioning

The learning of an association between a CS and a US can be retarded by unreinforced presentations of the CS alone (termed latent inhibition or LI) or by un-correlated presentations of the CS and US (termed learned irrelevance or LIRR). In rabbit eyeblink conditioning, there have been some recent failures to replicate LI. LIRR has been hypothesized as producing a stronger retardation effect than LI based on both empirical studies and computational models. In the work presented here, we examined the relative strength of LI and LIRR in eyeblink conditioning in rabbits and humans. In both species, a number of pre-exposure trials sufficient to produce LIRR failed to produce LI (Experiments 1 & 3). Doubling the number of CS pre-exposures did produce LI in rabbits (Experiment 2), but not in humans (Experiment 4). LI was demonstrated in humans only after manipulations including an increased inter-trial interval or ITI (Experiment 5). Overall, it appears that LIRR is a more easily producible pre-exposure retardation effect than LI for eyeblink conditioning in both rabbits and humans. Several theoretical mechanisms for LI including the conditioned attention theory, stimulus compression, novelty, and the switching theory are discussed as possible explanations for the differences between LIRR and LI. Overall, future work involving testing the neural substrates of pre-exposure effects may benefit from the use of LIRR rather than LI.     

Pharmacological dissociation of trace and long-delay fear conditioning in young rats

In most studies comparing trace and delay conditioning, CS duration is kept constant across training conditions but the interstimulus interval (ISI), the time from CS onset to US onset, is confounded. In the infrequently used long-delay condition, however, ISI is kept constant across the trace and delay conditions but CS duration varies. A recent study reported that trace and long-delay fear conditioning have the same developmental trajectory, with both emerging later in development than standard-delay conditioning (). Past studies have shown that trace conditioning is mediated by the cholinergic system; given the parallel developmental emergence of trace and long-delay conditioning, the present study examined whether the cholinergic system also mediates long-delay conditioning. Two experiments, both involving Sprague-Dawley-derived rats and using freezing as a measure of learned fear, showed that the cholinergic system is critically involved in trace conditioning but is not involved in long-delay conditioning. Specifically, pre-training injections of the muscarinic receptor antagonist scopolamine impaired acquisition of a CS-US association in 32-day-old rats trained with a trace procedure but had no effect on rats this age trained with a long-delay procedure (Experiment 1). Similarly, pre-training injections of physostigmine, a cholinesterase inhibitor, enhanced acquisition of trace conditioning in 25-day-old rats but had no effect on long-delay conditioning in rats this age (Experiment 2). Taken together, the results indicate that despite the similarities between trace and long-delay conditioning in terms of developmental emergence and level of conditioned responding, they are mediated by different physiological systems.     

A Comparison of Latent Inhibition and Learned Irrelevance Pre-Exposure Effects in Rabbit and Human Eyeblink Conditioning

The learning of an association between a CS and a US can be retarded by unreinforced presentations of the CS alone (termed latent inhibition or LI) or by un-correlated presentations of the CS and US (termed learned irrelevance or LIRR). In rabbit eyeblink conditioning, there have been some recent failures to replicate LI. LIRR has been hypothesized as producing a stronger retardation effect than LI based on both empirical studies and computational models. In the work presented here, we examined the relative strength of LI and LIRR in eyeblink conditioning in rabbits and humans. In both species, a number of preexposure trials sufficient to produce LIRR failed to produce LI (Experiments 1 & 3). Doubling the number of CS pre-exposures did produce LI in rabbits (Experiment 2), but not in humans (Experiment 4). LI was demonstrated in humans only after manipulations including an increased inter-trial interval or ITI (Experiment 5). Overall, it appears that LIRR is a more easily producible pre-exposure retardation effect than LI for eyeblink conditioning in both rabbits and humans. Several theoretical mechanisms for LI including the conditioned attention theory, stimulus compression, novelty, and the switching theory are discussed as possible explanations for the differences between LIRR and LI. Overall, future work involving testing the neural substrates of pre-exposure effects may benefit from the use of LIRR rather than LI.     

Two-phase model analysis of the effects of interstimulus interval and masking task in human aversive classical conditioning

Three levels (.5, 1.0, and 2.0 sec) of interstimulus interval (ISI) and presence vs absence of a masking task were manipulated in a human eyelid reflex conditioning situation. Through the use of a two-phase model of conditioning performance, it was concluded that (1) increasing the ISI and introducing a masking task increased the duration of Phase 1, (2) as ISI increased, the amount of conditioning decreased, (3) the trial-by-trial rate of change in response probability increased as a function of ISI, and (4) the masking task reduced operator limits. It was also noted that the typical ISI function is composed of rate effects at short ISis and conditioning limit effects at long ISIs and that the larger 0 at larger ISis does not follow from a model of the stimulus trace hypothesis.     

The extinction of an aversion: Role of the interval between non-reinforced presentations of the averted stimulus

A toxiphobia conditioning paradigm was used to examine the relation between the intertrial interval (ITI) and the extinction of an aversion. The design employed was based upon that developed by Davis (1970) to study the relation between the ITI and the habituation of a response. After conditioning, the conditioned stimulus (CS) was presented on two occasions at times T1 and T2, and this interval (the ITI) was varied across groups. However, the interval from the CS exposure at T2 to the extinction test was common for all groups. On the test, ITIs of 6 to 24 hr were found to have promoted more extinction than ITIs of 0.5 hr with odour (Experiments 1a and b) and flavour (Experiment 1c) CSs. Further, this facilitation of extinction by the long ITIs compared to the short ones was not due to differences in the intervals between the initial CS exposure at T1 and the test (Experiment 2). The final experiment examined the relation between the ITI and extinction when different CSs were presented at T1 and T2. A short interval between the presentation of CS1 at T1 and the presentation of CS2 at T2 was found to have facilitated the extinction of the aversion to CS2 compared either to a long interval between these presentations or to the presentation of CS2 in the absence of a prior CS1 presentation. The results were discussed in terms of the model developed by Wagner (1981).     

Posttraining prefrontal lesions impair jaw movement conditioning performance,but have no effect on accompanying heart rate changes

This experiment examined the role of the medial prefrontal cortex (mPFC) in regulating learned autonomic and somatomotor responses in rabbits using appetitive Pavlovian conditioning. Interstimulus interval (ISI) duration [i.e., the time between the onset of the conditioned stimulus (CS) and unconditioned stimulus (US)] was manipulated in order to determine whether ISI duration was related to the heart rate (HR) responses obtained during conditioning. Two groups received either a 1- or a 4-s ISI, with a tone as the CS and an intraoral pulse of water as the US. Another two groups received explicitly unpaired presentations of either the 1- or 4-s tone CS and water US. Few conditioned jaw movement (JM) or HR conditioned responses (CRs) were observed in the unpaired conditions. Significant JM conditioning was, however, elicited by the paired conditions, especially to the 4-s ISI. Consistent CS-evoked HR accelerations were observed in both ISI conditions. After five sessions of training, the mPFC was lesioned in half the animals. A separate group of paired animals received sham lesions. After surgical recovery, all animals received 3 days of postoperative training. During the first postoperative training session, JM CRs significantly declined in both groups with mPFC lesions in comparison to the groups with sham lesions. The mPFC lesions, however, did not affect the CS-evoked cardiac accelerations, which again occurred during postoperative training.     

Background stimuli and the inter-stimulus interval during pavlovian conditioning

An experiment was carried out to investigate the part played by background stimuli during Pavlovian conditioning. Groups of rats were presented with tone CSs and unavoidable footshock USs. Two components of the inter-stimulus interval, namely CS duration and the duration of a trace interval, were manipulated across groups. Subsequently subjects were tested for their reactions to background stimuli by being re-exposed to the training environment in the absence of both the CS and US. The degree to which rats failed to avoid this environment was found to relate to CS duration by an inverted U curve. The manipulation of the trace interval however did not significantly affect avoidance.     

Appetitive pavlovian-instrumental interactions: Effects of inter-stimulus interval and baseline reinforcement conditions

Experiment I manipulated two variables which appear to influence whether a signal for food enhances or suppresses food-rewarded instrumental performance: interstimulus interval (ISI) during classical conditioning and instrumental reinforcement schedule during testing. In two groups a 10-s conditioned stimulus (CS) and food were paired (10-s and 20- to 100-s ISI), while in a third group they were unpaired. During signalled reinforcement of lever-pressing (S+), the CS suppressed responding in both paired groups. During signalled extinction (S-), responding in the 10-s ISI group was suppressed during the CS and enhanced for 60 s after CS offset; responding in the 20- to 100-s ISI group was enhanced both during the CS and for 120 s after CS offset. Experiment II examined whether the long ISI enhancement effect would occur when the baseline response rate was lowered by satiation rather than extinction. A 20- to 100-s CS and food were paired in one group and unpaired in another. After near-satiation on a CRF schedule, CS presentations caused a reduction in responding in both groups, with no significant difference between the two groups. The results of the two experiments were interpreted in terms of an interaction between the expectancy of food generated by stimuli conditioned at short and long ISIs and the expectancy of food availability controlled by the instrumental schedule.     

Timed excitatory conditioning under zero and negative contingencies

Rats (rattus norvegicus) anticipated the arrival of a food pellet unconditioned stimulus (US) even when the conditioned stimulus (CS) signaled no overall change or a substantial decrease in the overall rate of US occurrence. Pellet USs were scheduled probabilistically in the intertrial interval at either an equivalent rate (Experiment 1) or a four times higher rate (Experiments 2 and 3) than in the CS, which included one fixed-time target US. Conditioning has been said to involve learning "whether" (contingency) the CS signals a change in the US, and if so, "when" (contiguity) the US is scheduled to arrive. Our results suggest that "when" trumps "whether," challenging the received view that a positive CS-US contingency is necessary for successful conditioning.     

The potentiation effect during serial conditioning

The development of suppression in rats to a target conditioned stimulus (CS) was compared in trace and serial conditioning procedures. The interval between the end of the target CS and the shock unconditioned stimulus (US) was filled by a second CS in the serial, but not the trace, procedure. In five experiments the serial procedure produced superior conditioning. This potentiation effect, however, depended critically upon the level of conditioning to the stimulus interpolated between the target CS and the US. When conditioning to the interpolated CS was either reduced by giving independent nonreinforced trials with this CS alone or enhanced by independent reinforced trials, the potentiation effect was abolished. In addition, the insertion of a trace interval between the target and interpolated CSs reduced the effect. However, the magnitude of conditioning to the target CS was unaffected by post-conditioning changes in the conditioned strength of the interpolated CS. These findings are discussed in terms of the contribution of both the association between the CSs themselves, which is inherent in the serial procedure, and that between the target CS and the US.     

The decrement in conditioned fear with increased trials of simultaneous conditioning is not specific to the simultaneous procedure

Three experiments using a conditioned punishment paradigm with rat subjects examined the possibility that the nonmonotonic acquisition function previously found to characterize simultaneous conditioning was due to the noninformative nature of the conditioned stimulus (CS). In Experiment 1 the suppressive effects of a CS previously presented with an unconditioned stimulus (US) in a simultaneous and forward (informative) manner were compared following 20 and an additional 60 conditioning trials. Excitatory conditioning similarly diminished with increased trials for both the simultaneous and forward procedures. Experiment 2 employed a between-groups design. Simultaneous, forward, and trace conditioning procedures were compared following 20 or 100 trials. Each of the three 100-trial groups showed less resistance to extinction than their 20-trial counterparts. Experiment 3 determined that the decrement in excitatory conditioning for the 100-trial groups was not due to the greater number of US presentations, per se, but rather to the number of CS-US pairings. The nonmonotonic acquisition function observed with all three conditioning procedures indicated that informational factors were not responsible for the decrement observed in simultaneous conditioning. The pattern of results suggested that subjects receiving extended conditioning trials were better able to discriminate between training and testing.     

Associative changes with a random CS-US relationship

Three experiments examined conditioned magazine approach in rats when a positive unconditioned stimulus (US) bore a random relation to a conditioned stimulus (CS). Experiment 1 found that over the course of conditioning the CS initially elevated responding relative to the baseline but then lost the power to do so. Transfer tests revealed that a CS-US association developed early and persisted despite the decline in magazine responding. Experiment 2 confirmed the persistence of CS-US associations and found them to be more substantial when a different US occurred during the CS than in its absence. In Experiment 3, when the situation was exposed to US-alone presentations prior to introducing the CS, there was little evidence that a subsequent random relation between the CS and US produced an association between them. These results agree with those of blocking and overshadowing experiments using discrete CSs and support an interpretation of the random procedure in terms of competition between the background and CS for conditioning.