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Real-time processing of serial stimuli in classical conditioning of the rabbit's nictitating membrane response.

Real-time models contend that a conditioned stimulus (CS) gives rise to a cascade of hypothetical stimuli that govern conditioned responses (CRs) on a moment-by-moment basis. Experiments with the rabbit nictitating membrane response successfully extended these models to external stimuli. CSs were trained in sequence with an unconditioned stimulus (CSA-CSB-US). When the CSA-CSB interval was shortened, the CR was compressed; when the CSA-CSB interval was lengthened, the CR was broadened. Peaks appeared at 2 places, namely, 1 following CSA by a period equal to its CS-US interval and another following CSB by its CS-US interval. Outside the sequence, the individual CSs evoked CRs located between their respective CS-US intervals. When, however, the 2 CSs were trained separately, the CRs were appropriate to their respective CS-US intervals when tested alone or in sequence. The results are discussed in terms of the J.E. Desmond and J.W. Moore (1988) and S. Grossberg and N.A. Schmajuk (1989) models.

Animals↗

Aging and classical conditioning: parallel studies in rabbits and humans.

The model system of classical conditioning of the eyelid response in rabbits has led to significant progress in the understanding of the neural circuitry involved in learning and memory. We are now at a point where we can begin to explore how aging in the neural circuitry affects this basic form of associative learning and memory over the life span. Parallels between humans and animals in aging and classical conditioning of the eyelid response are identified. A hypothetical model describing how acquisition occurs in the cerebellum has proved useful in predicting where in the cerebellum age changes might be most likely to affect classical conditioning. The loss of Purkinje cells with age is one of the cerebellar age changes likely to affect classical conditioning in aged mammals. Purkinje cell loss is only one of several age changes in the cerebellum which may impair acquisition and retention of the conditioned eyeblink response. This model system is of demonstrated utility in extending understanding of the neurobiology of learning, memory, and aging in humans as well as animals.

Adult↗

Classical conditioning of insulin effects in healthy humans.

OBJECTIVE: Classical conditioning of insulin effects was examined in healthy humans using a placebo-controlled design. This study examined whether subjects who experienced a conditioned stimulus (CS) paired with insulin in the acquisition phase of a conditioning protocol would show a conditioned decrease of blood glucose when receiving the CS with a placebo injection in the test phase. METHODS: Twenty healthy male students were assigned either to group 1, which received insulin (0.035 IU/kg i.v.), or to group 2, which received i.v. saline on 4 consecutive days (acquisition). On day 5 (test), both groups were injected with saline. The CS was an olfactory stimulus. Blood glucose, serum insulin, plasma glucagon, plasma catecholamines, serum cortisol, and symptoms were repeatedly measured during each session. RESULTS: In the test phase, group 1 reacted with a significantly larger decrease of blood glucose after presentation of the CS than group 2. Within group 1, a larger conditioned blood glucose decrease was associated with features that enhance classical conditioning (ie, intensity of the unconditioned response and intensity of the CS). Furthermore, in group 1, there was an increase of baseline insulin from day 1 to day 5 and a tendency for insulin reduction after CS presentation. Groups also tended to differ in cortisol and neuroglycopenic symptoms after CS presentation. CONCLUSIONS: Conditioned effects in blood glucose are in accordance with the predictions. As a result of the exploratory analyses, our data also provide hints about conditioned changes in insulin, counterregulatory hormones, and symptoms.

Adult↗

An experimental analysis of classically conditioned nausea during cancer chemotherapy.

This study investigated classical conditioning in women undergoing outpatient adjuvant chemotherapy for breast cancer. Breast cancer chemotherapy outpatients were randomly assigned either to an Experimental Group (exposed to a distinctive stimulus before each infusion of chemotherapy) or to a Control Group. After repeated infusions of chemotherapy, patients' responses to the experimental stimulus were assessed in a location not associated with chemotherapy. Experimental Group patients had increased nausea (self-reported on a visual analog scale) following the presentation of the experimental stimulus at this test trial, whereas Control Group patients did not. Two other measures of nausea corroborated these results. Post hoc statistical analyses confirmed predictions based on conditioning theory. This conditioning model of anticipatory nausea bears witness to the relevance of classical conditioning in clinical medicine.

Antineoplastic Combined Chemotherapy Protocols↗

Conditioning of skin conductance levels through a combined instrumental-classical conditioning procedure.

A paucity of studies and inconsistent results characterize attempts to instrumentally condition tonic electrodermal activity. The present study was conducted to further explore the classical and instrumental conditioning of skin conductance levels (SCLs) through auditory analogue feedback. Fifty undergraduates were randomly assigned to one of five groups of equal size: instrumental conditioning alone, classical conditioning alone, instrumental and classical conditioning combined, and two control groups. The combined training group achieved significantly higher SCLs relative to the group receiving instrumental conditioning alone.

Analysis of Variance↗

No evidence of classical conditioning of electrodermal responses during anesthesia.

Classical conditioning of skin conductance responses was studied in 26 healthy volunteers and 31 patients who received general anesthesia (0.5 MAC isoflurane-70% nitrous oxide) for minor gynecologic surgery to determine whether there was autonomic evidence of new learning during anesthesia. In the conditioning phase, a loud noise was paired with a target word (conditioned stimulus [CS]). Conditioning was established if the CS produced conditioned responses, i.e., greater skin conductance responses than other, "filler words." In the subsequent elicitation phase, we investigated whether it was possible to elicit similar conditioned responses to the CS when it was presented without noise about 3 h later. Conditioning was established in the control group, who received no anesthesia. There were differences in response magnitudes of 0.32 and 0.27 mu siemen between responses to the CS and filler words, P less than 0.001. In contrast, there was no evidence that conditioning was established during anesthesia. In the elicitation phase, conditioned responses to the CS could be elicited in volunteers; i.e., response magnitudes were greater for the CS than other words (during the first two blocks of trials), with differences of 0.19 to 0.48 mu siemen, P less than 0.05 to P less than 0.001. In contrast, responses to the CS could not be elicited in patients after anesthesia. There was also no evidence of recognition of words that were presented during anesthesia in patients, but recognition was manifested in volunteers. Thus, in contrast to our previous demonstration of conditioning in subjects who received 30% nitrous oxide in oxygen, there was no evidence of conditioning in anesthetized patients.

Adult↗

[Modulatory role of the cerebellar cortex during the classically conditioned nictitating membrane response in rabbit].

Classical conditioning of the rabbit nictitating membrane(NM) response was accomplished by standard delay procedure of paired tone (CS) and airpuff (US) stimuli. Upon reaching a criterion of 90% conditioned responses (CRs) in 3 consecutive blocks, an electrolytic lesion was made at the hemicerebellar VI cortical lobule on the training side. Results showed that the learned response of the dentate-interpositus (D-I) nuclei and the conditioned NM response were completely or almost completely eliminated without affecting the unconditioned response. However, cerebellar cortical lesion performed on animals being over 1 week overtrained produced no effect on CRs and the learned response of the D-I nuclei. Meanwhile, the spontaneous activity of the D-I nuclei during consolidation reduced in frequency as a result of the cortical lesion at the early stage of learning. Our findings indicate that the cerebellar cortex plays a modulatory role in the generation of the conditioned NM response and the development of the learned response of the D-I nuclei at an early stage of learning. With consolidation, the D-I nuclei become free of control from the cerebellar cortex and serve as the essential component of the memory trace for this model system.

Animals↗

Lesions of the middle cerebellar peduncle disrupt acquisition and retention of the rabbit's classically conditioned nictitating membrane response.

Rabbits were classically conditioned to emit a nictitating membrane response (NMR) to either a light or tone conditioned stimulus (CS) paired with an eye shock unconditioned stimulus (UCS). They then received lesions of the middle cerebellar peduncle (MCP) or served as unoperated controls. Following surgery, they were given separate presentations of tone, light, and vibratory CSs, each paired with the eye shock UCS. In this way, conditioned responses (CR) to the previously trained light or tone served as a test of retention, whereas CRs to the remaining two conditioned stimuli (tone and vibratory or light and vibratory) served as a test of acquisition. The results of the study revealed that rabbits with complete lesions of the MCP showed disrupted acquisition and retention of the conditioned NMR to all stimuli, rabbits with partial MCP lesions also showed disrupted acquisition and retention to all CSs, but to a lesser degree, and animals with lesions that missed the MCP and unoperated controls both showed normal acquisition and retention of the conditioned NMR. These data are consistent with the view that the cerebellum is an essential part of the circuit for classical conditioning of the NM response and that information about CSs in the auditory, visual, and tactile modalities reach the cerebellum by way of the MCP.

Afferent Pathways↗

Classical conditioning in Aplysia californica.

A form of aversive classical conditioning is described in which a chemosensory conditioned stimulus rapidly acquires the ability to modulate a defensive response (escape locomotion). Because Aplysia show both sensitization and classical conditioning, it is now possible to begin to examine the relationship between nonassociative and associative learning on behavioral and cellular levels.

Animals↗

In vitro classical conditioning of abducens nerve discharge in turtles.

In vitro classical conditioning of abducens nerve activity was performed using an isolated turtle brainstem-cerebellum preparation by direct stimulation of the cranial nerves. Using a delayed training procedure, the in vitro preparation was presented with paired stimuli consisting of a 1 sec train stimulus applied to the auditory nerve (CS), which immediately preceded a single shock US applied to the trigeminal nerve. Conditioned and unconditioned responses were recorded in the ipsilateral abducens nerve. Acquisition exhibited a positive slope of conditioned responding in 60% of the preparations. Application of unpaired stimuli consisting of CS-alone, alternate CS and US, or backward conditioning failed to result in conditioning, or resulted in extinction of CRs. Latencies of CR onset were timed such that they occurred midway through the CS. Activity-dependent uptake of the dye sulforhodamine was used to examine the spatial distribution of neurons labeled during conditioning. These data showed label in the cerebellum and red nucleus during conditioning whereas these regions failed to label during unconditioned responses. Furthermore, the principal abducens nucleus labeled heavily during conditioning. These findings suggest the feasibility of examining classical conditioning in a vertebrate in vitro brainstem-cerebellum preparation. It is postulated that the abducens nerve CR represents a behavioral correlate of a blink-related eye movement. Multiple sites of conditioning are hypothesized, including the cerebellorubral circuitry and brainstem pathways that activate the principal abducens nucleus.

Abducens Nerve↗

Retention of a classically conditioned reflex response in spinal cat.

Retention of classically conditioned flexion reflex facilitation was examined in unanesthetized, decerebrate, acute spinal cats. Flexion reflex facilitation, recorded from the tibialis anterior muscle, was obtained by pairing saphenous nerve stimulation (the conditioned stimulus) with superficial peroneal nerve stimulation (the unconditioned stimulus). The flexion reflex declined in control animals receiving the same number of nerve stimuli over the same time span, but in an explicitly unpaired sequence. To investigate retention, conditioned stimuli were presented at 5-min intervals following acquisition for a 2 1/2-h period. During this time a significant difference between conditioning and control groups was maintained even to the last trial, with no indication that the difference was subsiding over time. The results support the possibility that a classical conditioning paradigm applied to the spinal cord can induce alterations in spinal reflexes of long duration. Furthermore, the results appear to rule out post-tetanic potentiation as a mechanism producing the observed long-term effects.

Animals↗

Learning of physiological responses: II. Classical conditioning of the baroreflex.

The baroreflex can be classically conditioned. In neuromuscular blocked (NMB) rats, electrical stimulation of the aortic depressor nerve (ADN) and dopamine-produced blood pressure rise were effective unconditioned stimuli (UCS) for auditory discriminative classical conditioning. The conditioned response (CR) pattern (bradycardia, vasodilatation, and hypotension > 10 torr) closely resembled that of the unconditioned baroreflex. Conditioned stimulus (CS) specificity was demonstrated by discrimination of baroreflex-associated and nonassociated auditory stimuli, and also by elaborating depressor and pressor CRs to auditory CSs, which respectively had been associated with either baro-afferent (depressor) or tail-shock (pressor) UCSs. The conditioned-baroreflex-magnitude increased with trials. These findings support quantitative models in which CRs interact with and calibrate the gain and dynamic properties of natural reflexes.

Acoustic Stimulation↗

Conditioned stimulus intensity: role of cutaneous fiber size in classical conditioning of the flexion reflex in the spinal cat.

Three intensities of the conditioned stimulus (CS) were used to investigate the role of cutaneous fiber size on classical conditioning of the flexion reflex in the spinal cat. Conditioning occurred when the CS activated A alpha and A delta cutaneous fibers; CS activation of only the largest myelinated (A alpha) fibers or CS recruitment of unmyelinated (C) fibers in addition to A alpha and A delta fibers did not lead to significant conditioning. Comparison with two unpaired CS-US control treatment groups showed that the facilitation of reflex output during conditioning was specific to pairing of the CS and US and was not a consequence of sensitization or differential habituation rates between paired (conditioning) and unpaired (control) groups. These results were comparable to those from intact animal conditioning studies and thus lend further support to the spinal conditioning preparation as an appropriate simplified system for investigations of neural mechanisms of learning.

Animals↗

Fear is not critical to classically conditioned analgesia: the effects of periaqueductal gray lesions and administration of chlordiazepoxide.

A high correlation between fear and analgesia classically conditioned to footshock in rats has been reported in the literature. However, it has never been directly tested whether or not fear is in fact causal to the production of conditioned analgesia. We therefore tested whether conditioned analgesia could be elicited in the absence of fear by employing two independent methods of fear suppression. First, areas of the periaqueductal gray (PAG) previously implicated in fear were selectively lesioned. Lesions of the dorsolateral PAG significantly attenuated conditioned analgesia and markedly decreased fear responses. Second, fear was attenuated via administration of chlordiazepoxide (CDP). Rats which had been conditioned while in the presence of CDP showed no reduction in conditioned pain inhibition. These results demonstrate that: (1) fear is not causal to classically conditioned analgesia and (2) the anatomical substrates for fear and conditioned analgesia are distinct but partially overlapping. The fact that fear is not a critical antecedent for classically conditioned analgesia suggests that classical conditioning techniques may be applied clinically to increase the effectiveness of some analgesic manipulations.

Animals↗

Standard delay eyeblink classical conditioning is independent of awareness.

P. F. Lovibond and D. R. Shanks (2002) suggested that all forms of classical conditioning depend on awareness of the stimulus contingencies. This article considers the available data for eyeblink classical conditioning, including data from 2 studies (R. E. Clark, J. R. Manns, & L. R. Squire, 2001; J. R. Manns, R. E. Clark, & L. R. Squire, 2001) that were completed too recently to have been considered in their review. In addition, in response to questions raised by P. F. Lovibond and D. R. Shanks, 2 new analyses of data are presented from studies published previously. The available data from humans and experimental animals provide strong evidence that delay eyeblink classical conditioning (but not trace eyeblink classical conditioning) can be acquired and retained independently of the forebrain and independently of awareness. This conclusion applies to standard conditioning paradigms; for example, to single-cue delay conditioning when a tone is used as the conditioned stimulus (CS) and to differential delay conditioning when the positive and negative conditioned stimuli (CS+ and CS-) are a tone and white noise.

Aged↗

Classical conditioning in the fetal rat with a long delay between presentation of CS and US.

A single paired presentation of the artificial nipple and milk results in classical conditioning of changes in perioral responsiveness in the E20 rat fetus. This classical conditioning is evidenced by a reduction in responding to perioral tactile stimulation. The results of Experiment 1 confirmed the specificity of milk as an unconditioned stimulus to support classical conditioning. Experiment 2 demonstrated that single-trial classical conditioning with the artificial nipple CS and milk US was possible with a delay of 30 s between nipple and milk presentations. Further, measurements of fetal motor behavior during the delay between CS and US presentations indicated that a single 15-s presentation of the artificial nipple increased movements of the mouth for 30 s after removal of the artificial nipple. Experiment 3 demonstrated that three exposures to the artificial nipple prolonged the expression of mouthing for up to 120 s and made possible single trial classical conditioning with a delay of 120 s between CS and US presentations. The capacity of the fetal CNS to maintain a "behavioral trace" for an ecologically important stimulus, such as the nipple, could have adaptive significance in the early development of motivated behavior.

Animals↗

Classical conditioning of feeding in Aplysia: II. Neurophysiological correlates.

Feeding behavior in Aplysia californica can be classically conditioned using tactile stimulation of the lips as conditional stimulus (CS) and food as unconditional stimulus (US) [ (companion paper)]. Conditioning resulted in an increase in the number of CS-evoked bites that persisted for at least 24 hr after training. In this study, neurophysiological correlates of classical conditioning training were identified and characterized in an in vitro preparation of the cerebral and buccal ganglia. Stimulation of a lip nerve (AT(4)), which mediates mechanosensory information, resulted in a greater number of buccal motor patterns (BMPs) in ganglia isolated from animals that had received paired training than in ganglia from control animals. The majority of the evoked BMPs were classified as ingestion-like patterns. Intracellular recordings from pattern-initiating neuron B31/32 revealed that stimulation of AT(4) evoked greater excitatory input in B31/32 in preparations from animals that had received paired training than from control animals. In contrast, excitatory input to buccal neuron B4/5 in response to stimulation of AT(4) was not significantly increased by paired training. Moreover, correlates of classical conditioning were specific to stimulation of AT(4). The number of spontaneously occurring BMPs and the intrinsic properties of two buccal neurons (B4/5 and B31/32) did not differ between groups. These results suggest that appetitive classical conditioning of feeding resulted in the pairing-specific strengthening of the polysynaptic pathway between afferent fibers and pattern-initiating neurons of the buccal central pattern generator.

Action Potentials↗

Activity-dependent enhancement of presynaptic facilitation provides a cellular mechanism for the temporal specificity of classical conditioning in Aplysia.

A hallmark of many forms of classical conditioning is a precise temporal specificity: Learning is optimal when the conditioned stimulus (CS) slightly precedes the unconditioned stimulus (US), but the learning is degraded at longer or backward intervals, consistent with the notion that conditioning involves learning about predictive relationships in the environment. To further examine the cellular mechanisms contributing to the temporal specificity of classical conditioning of the siphon-withdrawal response in Aplysia, we paired action potential activity in siphon sensory neurons (the neural CS) with tail nerve shock (the US) at three critical time points. We found that CS-US pairings at short (0.5 sec) forward intervals produced greater synaptic facilitation at sensorimotor connections than did either 0.5-sec backward pairings or longer (5 sec) forward pairings, as reflected in a differential increase in both the amplitude and rate of rise of the synaptic potential. In the same preparations, forward pairings also differentially reduced the sensory neuron afterhyperpolarization relative to backward pairings, suggesting that changes in synaptic efficacy were accompanied by temporally specific changes in ionic currents in the sensory neurons. Additional experiments demonstrated that short forward pairings of sensory cell activity and restricted applications of the neuromodulatory transmitter serotonin (normally released by the US) differentially enhanced action potential broadening in siphon sensory neurons, relative to backward pairings. Taken together, these results suggest that temporally specific synaptic enhancement engages both spike-width-dependent and spike-width-independent facilitatory processes and that activity-dependent enhancement of presynaptic facilitation may contribute to both the CS-US sequence and proximity requirements of conditioning.

Animals↗