The role of awareness in delay and trace fear conditioning in humans

Expression of conditional fear without awareness has been previously demonstrated during delay conditioning, a procedure in which the conditioned stimulus (CS) and unconditioned stimulus (UCS) overlap. However, less is known about the role of awareness in trace fear conditioning, where an interval of time separates the CS and UCS. The present study assessed skin conductance response (SCR) and UCS expectancy during delay and trace conditioning. UCS predictability was varied on a trial-bytrial basis by presenting perithreshold auditory CSs. Differential UCS expectancies were demonstrated only on perceived delay and trace trials. Learning-related SCRs were observed during both perceived and unperceived delay CSs. In contrast, differential SCRs were demonstrated only for perceived trace CSs. These data suggest that awareness is necessary for conditional responding during trace, but not delay, fear conditioning.     

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.     

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.     

Human trace fear conditioning: right-lateralized cortical activity supports trace-interval processes

Pavlovian conditioning requires the convergence and simultaneous activation of neural circuitry that supports conditioned stimulus (CS) and unconditioned stimulus (US) processes. However, in trace conditioning, the CS and US are separated by a period of time called the trace interval, and thus do not overlap. Therefore, determining brain regions that support associative learning by maintaining a CS representation during the trace interval is an important issue for conditioning research. Prior functional magnetic resonance imaging (fMRI) research has identified brain regions that support trace-conditioning processes. However, relatively little is known about whether this activity is specific to the trace CS, the trace interval, or both periods of time. The present study was designed to disentangle the hemodynamic response produced by the trace CS from that associated with the trace interval, in order to identify learning-related activation during these distinct components of a trace-conditioning trial. Trace-conditioned activity was observed within dorsomedial prefrontal cortex (PFC), dorsolateral PFC, insula, inferior parietal lobule (IPL), and posterior cingulate (PCC). Each of these regions showed learning-related activity during the trace CS, while trace-interval activity was only observed within a subset of these areas (i.e., dorsomedial PFC, PCC, right dorsolateral PFC, right IPL, right superior/middle temporal gyrus, and bilateral insula). Trace-interval activity was greater in right than in left dorsolateral PFC, IPL, and superior/middle temporal gyrus. These findings indicate that components of the prefrontal, cingulate, insular, and parietal cortices support trace-interval processes, as well as suggesting that a right-lateralized fronto-parietal circuit may play a unique role in trace conditioning.     

Pavlovian inhibitory conditioning and tolerance to pentobarbital-induced hypothermia in rats

In this experiment we investigated inhibitory Pavlovian conditioning in the development of tolerance to pentobarbital-induced hypothermia. During an initial phase, one group of rats (discrimination group) received training in which, on alternate days, one conditional stimulus (CS+) was associated with administration of 30 mg/kg pentobarbital, and a different conditional stimulus (CS-) was associated with administration of physiological saline. During the phase, control groups received either exposure to both CSs but not the drug or to the drug but no CSs or to neither the CSs nor the drug. Subsequently, half the rats in each group received injections of pentobarbital in the presence of one of the CSs and the remaining half in the presence of the other CS. Rats from the discrimination group injected with pentobarbital in the presence of CS+ displayed the most tolerance (i.e., smallest drug effect), whereas rats from the discrimination group injected with pentobarbital in the presence of CS- displayed the least tolerance (i.e., greatest drug effect). The attenuation of tolerance seen in rats of the discrimination group injected in the presence of CS- provides evidence of inhibitory Pavlovian conditioning. Additional evidence of inhibitory conditioning was provided by the fact that CS2 enhanced the hypothermic effects of pentobarbital in the discrimination group, whereas CS1 attenuated these effects. Implications of the results for the nature of inhibitory conditioning are discussed.     

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.     

Concomitant Pavlovian conditioning of heart rate and leg flexion responses in the rat

Pavlovian conditioning was studied in male Fischer 344 rats using tones as the conditioned stimulus (CS) and footshock as the unconditioned stimulus (UCS). Different groups of animals received (a) contiguous CS-UCS pairings with a 0.5 sec CS, (b) contiguous CS-UCS pairings with a 4.0 sec CS, or (c) a random sequence of noncontiguous tones and shocks using either a 0.5 sec or a 4.0 sec CS. Heart rate (HR) and leg flexion (LF) responses were recorded. Leg flexion conditioning occurred only in the 0.5 sec contiguous group. Decelerative HR CRs occurred only in the 4.0 sec contiguous group. Accelerative HR changes occurred in the other two groups but were significantly greater in the 0.5 sec contiguous group. These results are similar to but not identical to those obtained during eyeblink or nictitating membrane conditioning in rabbits, and suggest that the topography of the Pavlovian HR CR is dependent on the simultaneous occurrence of other classically conditioned responses.     

Undernutrition during the brain growth period of the rat significantly delays the development of processes mediating Pavlovian trace conditioning

Pavlovian trace conditioning procedures allow one to assess the ability of animals to associate events that are temporally separated because the conditioned stimulus terminates prior to the occurrence of the unconditioned stimulus. We report that undernutrition during Postnatal Days 2-18 significantly delays the development of trace conditioning to a visual stimulus. Previously undernourished 20-day-old rats conditioned when no temporal interval separated the termination of the CS and US onset (0-s trace interval). However, it was not until the undernourished pups were 30 days old that they conditioned when the trace interval was 10 or 30 s. In contrast, control pups only 20 days old were able to condition when a 10-s trace interval separated the CS and US events, and 25-day-old control pups conditioned when the interval was either 10 or 30 s. These results suggest that undernutrition delays the development of processes that enable the rat to sustain a representation of a visual CS during the trace interval.     

Timing of conditioned responses utilizing electrical stimulation in the region of the interpositus nucleus as a CS

A large body of evidence indicates that the cerebellum is essential for the acquisition, retention, and expression of the standard delay conditioned eyeblink response and that the basic memory trace appears to be established in the anterior interpositus nucleus (IP). Adaptive timing of the conditioned response (CR) is a prominent feature of classical conditioning—the CR peaks at the time of onset of the unconditioned stimulus (US) over a wide range of CS-US interstimulus intervals (ISI). A key issue is whether this timing is established by the cerebellar circuitry or prior to the cerebellum. In this study timing of conditioned eyeblink responses established via electrical stimulation of the interpositus nucleus as a conditioned stimulus (CS) was analyzed prior to and following modification of the CS-US interval in well-trained rabbits. Consistent with previous results, learning under these conditions is very rapid and robust. The CR peak eyeblink latencies are initially timed to the US onset and adjust accordingly to lengthening or shortening of the CS-US interval, just as with peripheral CSs. The acquisition of conditioned eyeblink responses by direct electrical stimulation of the IP as a CS thus retains temporal flexibility following shifts in the CS-US delay, as found in standard classical eyeblink conditioning procedures.     

Timing of Conditioned Responses Utilizing Electrical Stimulation in the Region of the Interpositus Nucleus as a CS

A large body of evidence indicates that the cerebellum is essential for the acquisition, retention, and expression of the standard delay conditioned eyeblink response and that the basic memory trace appears to be established in the anterior interpositus nucleus (IP). Adaptive timing of the conditioned response (CR) is a prominent feature of classical conditioning-the CR peaks at the time of onset of the unconditioned stimulus (US) over a wide range of CS-US interstimulus intervals (ISI). A key issue is whether this timing is established by the cerebellar circuitry or prior to the cerebellum. In this study timing of conditioned eyeblink responses established via electrical stimulation of the interpositus nucleus as a conditioned stimulus (CS) was analyzed prior to and following modification of the CS-US interval in well-trained rabbits. Consistent with previous results, learning under these conditions is very rapid and robust. The CR peak eyeblink latencies are initially timed to the US onset and adjust accordingly to lengthening or shortening of the CS-US interval, just as with peripheral CSs. The acquisition of conditioned eyeblink responses by direct electrical stimulation of the IP as a CS thus retains temporal flexibility following shifts in the CS-US delay, as found in standard classical eyeblink conditioning procedures.     

Learning in cod (Gadus morhua): long trace interval retention

Basic knowledge about learning capacities and awareness in fish is lacking. In this study we investigated which temporal gaps Atlantic cod could tolerate between two associated events, using an appetitive trace-conditioning paradigm with blinking light as conditioned stimulus (CS) and dry fish food as unconditioned stimulus (US). CS–US presentations were either temporally overlapping (delay conditioning, CS duration 24 s, interstimulus interval 12 s) or separated by 20, 60, or 120 s (trace conditioning, CS duration 12 s) or 2 h (control, CS duration 12 s). The percentage of fish in the feeding area increased strongly during CS presentation in all delay, 20 s, and 60 s trace groups and in one out of two 120 s trace groups, but not in the control groups. In the 20 and 60 s trace procedures, the fish crowded together in the small feeding area during the trace interval, showing strong anticipatory behaviour. In all the conditioned groups, the fish responded to the CS within eight trials, demonstrating rapid learning. At 88 and 70 days after the end of the conditioning experiments, the delay and 20 s trace groups, respectively, were presented the CS six times at 2-h intervals without reward. All groups responded to the light signal, demonstrating memory retention after at least 3 months. This study demonstrates that Atlantic cod has an impressively good ability to associate two time-separated events and long time retention of learnt associations.     

Overshadowing by fixed- and variable-duration stimuli

Two experiments investigated the effect of the temporal distribution form of a stimulus on its ability to produce an overshadowing effect. The overshadowing stimuli were either of the same duration on every trial, or of a variable duration drawn from an exponential distribution with the same mean duration as that of the fixed stimulus. Both experiments provided evidence that a variable-duration stimulus was less effective than a fixed-duration cue at overshadowing conditioning to a target conditioned stimulus (CS); moreover, this effect was independent of whether the overshadowed CS was fixed or variable. The findings presented here are consistent with the idea that the strength of the association between CS and unconditioned stimulus (US) is, in part, determined by the temporal distribution form of the CS. These results are discussed in terms of time-accumulation and trial-based theories of conditioning and timing.     

Temporal coding in conditioned inhibition: analysis of associative structure of inhibition

Two experiments with rats as subjects were conducted to investigate the associative structure of temporal control of conditioned inhibition through posttraining manipulation of the training excitor-unconditioned stimulus (US) temporal relationship. Experiment 1 found that following simultaneous Pavlovian inhibition training (i.e., A --> US/XA-no US) in which a conditioned stimulus (CS A) was established as a delay excitor, maximal inhibition was observed on a summation test when CS X was compounded with a delay transfer CS. Furthermore, posttraining shifts in the A-US temporal relationship from delay to trace resulted in maximal inhibition of a trace transfer CS. Experiment 2 found complementary results to Experiment 1 with an A-US posttraining shift from serial to simultaneous. These results suggest that temporal control of inhibition is mediated by the training excitor-US temporal relationship.     

Aversive CS control of instrumental avoidance as a function of selected parameters and method of Pavlovian conditioning

Effects of aversive CSs upon dogs' instrumental avoidance responding were assessed as a function of the Pavlovian conditioning parameters using off-baseline (Experiment 1) and on-baseline (Experiment 2) conditioning procedures. In each experiment, three UCS intensitites (2, 6, and 10 mA) were crossed factorially with two CS-UCS intervals (10 and 90 sec) yielding six groups. After 4 days of conditioning, the CS-produced effects on avoidance were determined primarily by an interaction between Pavlovian procedure (off- versus on-baseline) and UCS intensity. With off-baseline conditioning, CS-produced facilitation of avoidance was an increasing function of the UCS intensity. However, with on-baseline conditioning, only the CS paired with weak UCS facilitated avoidance, and the CS paired with the strong UCS suppressed responding. This reversing parametric function poses problems for two-process motivational theories of avoidance.     

Temporal landmarks: proximity prevails

Subjects in conditioning experiments time their conditioned responses relative to the onsets of the conditioned stimuli (CSs). These onsets are temporal landmarks, by reference to which subjects may estimate the location of the unconditioned stimulus (US) in time. In a serial compound conditioning paradigm, a long duration CS comes on first, followed later by a second shorter CS, creating both a long-range and a short-range predictor of the US. We ask whether displacing the short-range predictor relative to the long-range predictor causes subjects to strike a compromise between the different temporal locations predicted by the two CSs. In three experiments with pigeons, we varied the training conditions so as to favor or militate against this outcome. However, in all conditions, there was no compromise; after the onset of the displaced short-range CS, the timing of conditioned responding was governed by it alone. This result contrasts with the compromises that are seen when the feeding time predicted by a CS is put in conflict with the time predicted by the circadian clock, and with the similar compromises sometimes seen when a nearby spatial landmark is displaced relative to a larger spatial context. Electronic Publication     

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.     

Summation of reinforcement rates when conditioned stimuli are presented in compound

Three experiments used delay conditioning of magazine approach in rats to examine the summation of responding when two conditioned stimuli (CSs) are presented together as a compound. The duration of each CS varied randomly from trial-to-trial around a mean that differed between the CSs. This meant that the rats' response rate to each CS was systematically related to the reinforcement rate of that CS, but remained steady as time elapsed during the CS (Harris & Carpenter, 2011; Harris, Gharaei, & Pincham, 2011). When the rats were presented with a compound of two CSs that had been conditioned separately, they responded more during the compound than during either of the CSs individually. More significantly, however, in all three experiments, the rats responded to the compound at the same rate as they responded to a third CS that had been reinforced at a rate equal to the sum of the reinforcement rates of the two CSs in compound. We discuss the implications of this finding for associative models (e.g., Rescorla & Wagner, 1972) and rate-based models (Gallistel & Gibbon, 2000) of conditioning.     

Analyzing the random control procedure: Effects of paired and unpaired CSs and USs on autoshaping the chick's key peck with heat reinforcement

A series of five experiments using a total of 264 subjects investigated the effects of paired and unpaired key light (CS) and heat (US) stimuli on autoshaping the chick's key peck. Experiment 1 established that paired presentations of CS and US promoted a more rapid rise in key pecking than did randomly presented CSs and USs and that the specific sequence of stimuli under the random control procedure affected key pecking performance. Experiment 2 used a trace conditioning procedure to determine the role of the CS-US interval on autoshaping and to define empirically unpaired CSs and USs. Key pecking declined as the trace delay interval was increased from 0 to 25 sec; at 25 sec, no conditioning of key pecking occurred. Experiments 3–5 assessed the effects on autoshaped key pecking of (a) number of daily CS-US pairings, (b) added unpaired CS presentations, and (c) added unpaired US presentations, since paired and random control schedules differed in all of these respects. Reduction in the number of CS-US pairings slowed the acquisition of key pecking as did the concurrent addition of nonreinforced CSs and unsignaled USs. These results support theories of association formation that stress the effects of both paired and unpaired CSs and USs.     

Effects of long- and variable-duration signals for food on activity,instrumental responding,and eating

Incentive-motivation theories typically assume that the conditioning of appetitive motivation involves the same parameters as Pavlovian salivary conditioning. In contrast, the Soltysik-Konorski model asserts that drive is inhibited by stimuli closely associated with food (salivary CSs) and augmented by stimuli more loosely associated with food (long and variable CS-US interval). Experiment 1 examined this latter proposition. Sixty-four rats were given extensive exposure to each of four environmental CSs, two while hungry and two while satiated. Within each deprivation condition, food was given 30–300 sec after placement of the rats in one environment, and was not given in the other environment. Performance on three separate measures—activity, lever-pressing, and food consumption—was higher in the environments previously associated with food. Experiment 2 examined the effects of discrete stimuli presented in advance of eating; in accord with the results of Experiment 1, food consumption was greater after a stimulus (1- to 9-min duration) previously paired with food than after no stimulus or after a stimulus unpaired with food. The overall results indicate (a) that stimuli associated with food become capable of facilitating a variety of food-directed behaviors, possibly via the conditioning of a common appetitive system, and (b) that a close association between the stimuli and food is not essential for such conditing to occur.     

A comparison of pecking generated by serial, delay, and trace autoshaping procedures

Pigeons were exposed to serial, delay, and trace autoshaping procedures. In Experiment I, all conditioned stimuli (CSs) were changes in illumination of the response key. The number of trials to acquisition of the keypeck increased from serial, to 4-sec delay, 8-sec delay, and 8-sec trace procedures, in that order. In Experiment II, which used a longer intertrial interval, trials to criterion increased from 8-sec delay, to 28-sec delay, 8-sec trace, and 28-sec trace procedures, in that order. In Experiment III, two groups received serial procedures in which the first CS was either a tone or a houselight, and the second was a keylight. The tone group acquired the key peck more rapidly than the houselight group. Early in conditioning in these experiments, and when the conditioned stimulus was a change in the keylight, there was a short latency to the onset of pecking and pecking was directed at the CS. After extensive conditioning, or when the CS was relatively diffuse, pecking still occurred, but had a longer latency and was not reliably directed toward the conditioned stimulus.