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A nonextinction procedure for long-term studies of classically conditioned enhancement of acoustic startle in the rat.

In the Fear-Potentiated Startle (FPS) paradigm, the reflexive response to a noise burst is enhanced when it is presented with a stimulus (typically a light) that previously had been paired with the presentation of an aversive stimulus, usually an electric shock. In the FPS paradigm, the conditioned effect is demonstrated under conditions of extinction, i.e., the light is not paired with the shock during FPS testing. Because of this, the FPS paradigm is of somewhat limited value as a longitudinal measure for studying classically conditioned enhancement of acoustic startle. The present studies report a simple and reliable nonshock procedure for studying classically conditioned potentiation of acoustic startle in the rat that does not utilize testing under conditions of extinction. Naive rats were exposed to 5-or 3-days/week startle test sessions for up to 20 weeks. Twenty (20) startle stimuli (115 dB noise bursts; 40 ms in duration) were presented during each session. Half of these startle stimuli were presented in darkness and half were immediately preceded by a 3500-ms presentation of a 15-W light. With this paradigm, the effects of pairing the light with the startle noise burst could be studied across many test sessions in the absence of extinction training. The light did not increase startle amplitude on the first few startle trials of the first test session. By the end of the first session, however, and continuing for many weeks of testing, startle responses in the presence of the light were significantly greater (by 30-40%) than in the absence of the light. The finding that the startle stimulus itself can serve as an unconditioned stimulus (UCS) to enhance subsequent startle responses replicates an earlier finding. The persistence of this Startle-Potentiated Startle (SPS) effect across multiple weeks of testing is in contrast to that earlier report.

Acoustic Stimulation↗

Classical conditioning model of panic disorder: response to Wolpe and Rowan.

We refute Wolpe and Rowan's (1989) classical conditioning theory of panic disorder based upon several lines of evidence presented above. While there is no doubt that hyperventilation and conditioning are important factors in the production of panic attacks, Wolpe and Rowan's (1989) hypothesis is inconsistent with recent data concerning the limited role of hyperventilation and the importance of cognitive mediation in the production of panic attacks.

Arousal↗

Hippocampal evoked potentials and EEG changes during classical conditioning in the rat.

Hippocampal evoked potentials (EPs) and EEG responses were studied in rats, using a classical conditioning paradigm (water, US), with a spatially discontiguous CS-US arrangement in order to separate the CS and goal-related responses. In early training, when the orienting score was high, the tone CS, instead of eliciting a definite EP, usually reset hippocampal theta activity in phase, i.e. theta rhythm became time-locked to CS. With further training, orienting activity (ORI) decreased to the preconditioning level, and this was associated with the recurrence of short-latency and high voltage hippocampal EPs, similar to those observed during habituation. This high voltage EP predicted that the animal would not orient any more towards CS. This correlation was confirmed by behavioural (satiation, shock US) and by pharmacological (scopolamine HBr, 2 mg/kg) treatments, all of which reduced the ORI score. Hippocampal EEGs also showed characteristic changes during conditioning. ORI towards CS was accompanied by higher frequency spectral peaks (9 c/sec) than response to US (7--8 c/sec). This correlation was seen both across sessions and within trials. We conclude that the above changes are related to orienting, attentional factors rather than to movement-related variables.

Animals↗

Impaired learning of classically conditioned bradycardia in rats following fourth ventricle administration of D-Ala2-methionine-enkephalinamide.

Prior to differential classical conditioning on two successive days, three groups of rats received an infusion (10 micrograms) of either the opioid peptide D-alanine2-methionine-enkephalinamide (DALA), DALA plus naltrexone (5 micrograms), or saline into the rostral region of the fourth ventricle. A fourth group, which served as a control to help localize DALA's site of action, received an infusion of DALA (10 micrograms) into the brain stem area on the floor of the ventricle. The group given DALA alone in the ventricle showed no evidence of a heart rate conditioned response (CR) either during conditioning or during a nondrug test session given 2 days after conditioning. Interference with the CR by DALA was reversed by the concomitant infusion of naltrexone. The control group given DALA in the brain stem developed a normal CR. It was suggested that DALA-induced opioid-receptor activity in the region of the periaqueductal/periventricular gray or locus coeruleus region of the ventricle may have prevented the learning of a CR. This could have occurred through a blunting of the emotional aftereffects of the unconditioned stimulus or through interference with projection pathways to other areas.

Animals↗

Altered activity in the hippocampus is more detrimental to classical conditioning than removing the structure.

Hippocampal ablation has no effect on the acquisition of the rabbit's classically conditioned nictitating membrane response. Systemic administration of scopolamine, which alters hippocampal neuronal activity, severely retards acquisition of the conditioned response in normal animals and those with cortical ablations. In animals with hippocampal ablations, however, scopolamine has no effect on conditioning. These findings suggest that altered neuronal activity in the hippocampus is more detrimental to conditioning than removing the structure.

Animals↗

Use of classical conditioning procedures to control aggressive behaviors in children: a preliminary report.

This experiment tested the effectiveness of classical conditioning procedures in reducing children's aggressive behaviors in a classroom setting. As hypothesized, 11 boys and 14 girls who were shown pictures of aggressive scenes paired with the presentation of negatively evaluated words subsequently displayed significantly fewer aggressive responses than controls who had not experienced these conditioning procedures. A significant teacher/classroom effect was also found, which appeared to be a function of the level of the 3 teachers' permissiveness. Specifically, children's aggressive behaviors occurred more frequently in the classroom where a permissive atmosphere prevailed than in the classroom where a more restrictive atmosphere was maintained. In those classrooms where a permissive atmosphere prevailed, the children who were the same sex as their teacher tended to display more aggressive responses than those children who were the opposite sex from their teacher.

Adolescent↗

Awareness of the CS-UCS contingency and classical conditioning of skin conductance responses with olfactory CSs.

The possibility of demonstrating acquisition of classically conditioned responses without awareness of the conditioned stimulus-unconditioned stimulus (CS-UCS) contingency using olfactory stimuli with 58 college student subjects was tested. A classical discrimination delay conditioning paradigm was employed, with electric shock as the UCS and two pleasant odors (perfumes) as the conditioned stimuli (CS+ and CS-). Trial-by-trial measures of skin conductance conditioned responses served as dependent variables. A masking task in the form of an olfactory memory task was employed for the purpose of delaying the onset of awareness of the conditioning contingency. Awareness of the conditioning contingency was assessed by a concurrent and a post hoc measure, and subjects who satisfied both criteria were considered aware of the CS-UCS contingency. Conditioning was observed only in the aware subjects, and only after the onset of awareness of the CS+-UCS contingency. Respiratory activity, measured as a check against possible artifacts, had no effect on the SCR measures. It was concluded that the awareness of the CS-UCS contingency is necessary for acquisition of discriminative conditioned responses in humans, regardless of the sensory modality in which CSs are presented. Sex differences in skin conductance measures and performance on the olfactory memory task were observed.

Adult↗

Classical conditioning and modification of the rabbit's (Oryctolagus cuniculus) unconditioned nictitating membrane response.

A fundamental tenet of behavior is that a reflex is automatic, unconscious, involuntary, and relatively invariant. However, we have discovered that a reflex can change dramatically as a function of classical conditioning, and this change can be demonstrated independently of the conditioned stimulus. We have termed this phenomenon conditioning-specific reflex modification (CRM). Although the behavioral laws and neural substrates of nonassociative reflex changes have been identified, the behavioral laws and neural substrates of CRM are only now being revealed. For example, CRM is similar to classical conditioning in that (a) it is a function of both the strength of conditioning and (b) the strength of the unconditioned stimulus, (c) it can be extinguished, and (d) it can be generalized from one unconditioned stimulus to another. Preliminary analysis suggests that CRM may have some features in common with post-traumatic stress disorder and may provide insights into treatment of the disorder.

Animals↗

Acquisition of conditional discriminations in hippocampal lesioned and decorticated rats: evidence for learning that is separate from both simple classical conditioning and configural learning.

This study examined whether hippocampal or neocortical lesions would impair acquisition of a discrimination task using taste aversions. Rats were injected with a drug 15 min before a flavored solution-lithium chloride pairing. On alternate days, vehicle injections preceded and followed access to the same flavored solution. Rats learned to consume significantly more of the flavored solution after vehicle injections than after drug injections. Rats with hippocampal lesions or neonatal decortication performed as well as controls. Rats with hippocampal lesions also learned a similar task in which visual and textural cues predicted whether access to a flavored solution would be followed by an injection of lithium chloride or vehicle. However, these hippocampal lesions did impair performance in the Morris water task. Occasion setting may involve a type of learning dissociated from both simple classical conditioning and configural learning.

Animals↗

MAPK signaling pathways mediate AMPA receptor trafficking in an in vitro model of classical conditioning.

The mitogen-activated protein kinase (MAPK) signal transduction pathways have been implicated in underlying mechanisms of synaptic plasticity and learning. However, the differential roles of the MAPK family members extracellular signal-regulated kinase (ERK) and p38 in learning remain to be clarified. Here, an in vitro model of classical conditioning was examined to assess the roles of ERK and p38 MAPK in this form of learning. Previous studies showed that NMDA-mediated trafficking of synaptic glutamate receptor 4 (GluR4)-containing AMPA receptors (AMPARs) underlies conditioning in this preparation and that this is accomplished through GluR4 interactions with the immediate-early gene protein Arc and the actin cytoskeleton. Here, it is shown that attenuation of conditioned responses (CRs) by ERK and p38 MAPK antagonists is associated with significantly reduced synaptic localization of GluR4 subunits. Western blotting reveals that p38 MAPK significantly increases its activation levels during late stages of conditioning during CR expression. In contrast, ERK MAPK activation is enhanced in early conditioning during CR acquisition. The results suggest that MAPKs have a central role in the synaptic delivery of GluR4-containing AMPARs during in vitro classical conditioning.

Animals↗

Postnatal development of respiratory and vocal responses during aversive classical conditioning in cats.

Respiratory rate, respiration amplitude, and vocal responses were recorded in cats of different ages during classic conditioning. Vocal responses to the conditional stimulus (CS) appeared first in 8-week-old kittens, and became prominent at older ages. An increase in respiration rate occurred after the onset of the CS in cats of all ages. Similarly, the decrease of respiration amplitude was observed at all ages, but only in 8-week-old and older subjects did this response resemble an adult pattern. In 4-week-old kittens the response was characterized by an early and brief peak, suggesting an alpha conditional response (CR). Clear and significant discrimination between a warning and a safety signal was present in both respiratory and vocal responses after the age of 8 weeks.

Aging↗

Classical conditioned response of rectosigmoid motility and regional cerebral activity in humans.

The relationship between the central processes of classical conditioning and conditioned responses of the gastrointestinal function is incompletely understood in humans. We tested the hypothesis that the rectosigmoid motility becomes conditioned with anticipatory painful somatosensory stimulus and that characteristic brain areas become activated during anticipation. In nine right-handed healthy male subjects, a loud buzzer (CS, conditional stimulus) was paired with painful transcutaneus electrical nerve stimulation to the right hand (unconditional stimulus). Rectosigmoid muscle tone measured by the barostat as the intrabag volume, phasic contractions of the bowel measured as the number of phasic volume events (PVEs), and regional cerebral blood flow assessed by positron emission tomography (PET), were measured before and after conditioning. Following conditional trials, the bag volume after CS alone did not show significant changes between before and after the stimulus, but the number of PVEs after 2-minute interval of the CS alone was significantly greater than that before the stimulus (P < 0.05). The PET data showed the conditioning elicited significant cerebral activation of the prefrontal, anterior cingulate, parietal and insula cortices (P < or = 0.001, uncorrected). Rectosigmoid motility can be conditioned with increase in phasic contractions in humans.

Adult↗

Inhibiting the expression of a classically conditioned behavior prevents its extinction.

The underlying neuronal substrates and behavioral properties that might mediate extinction of the classically conditioned eye-blink response (CR) were examined. Four groups of rabbits were trained to perform the CR. Two of the groups then received either three or six sessions of tone-alone extinction training while the motor nuclei that mediate expression of the CR (facial nucleus and accessory abducens) were reversibly inactivated with microinjections of the GABA agonist muscimol. After these inactivation extinction sessions, rabbits received four more extinction sessions without inactivation. Two groups of controls received either three or six extinction sessions while saline vehicle was infused into the motor nuclei, followed by four sessions with no infusions. Saline infusions had no effect on extinction, and controls extinguished the CR normally over the first three to four sessions. In contrast, muscimol inactivation of the motor nuclei completely prevented any performance of CRs during the three or six inactivation extinction sessions. At the start of the four extinction sessions without inactivation, rabbits performed CRs at the same rate and amplitude as controls on their first extinction sessions. The muscimol rabbits then extinguished the CR normally over the four sessions without inactivation. In short, inactivation of the motor nuclei completely prevented any extinction of the eye-blink CR with no effect on subsequent extinction without inactivation. These results are discussed in terms of possible neuroanatomical loci that might mediate the extinction process as well as how effects of manipulating CR performance during extinction may affect the extinction process.

Acoustic Stimulation↗

Immediate-early gene-encoded protein Arc is associated with synaptic delivery of GluR4-containing AMPA receptors during in vitro classical conditioning.

The immediate-early gene Arc is rapidly expressed in response to neuronal activity and is thought to be involved in mechanisms of synaptic plasticity. The function of Arc in these processes remains unknown. The present study demonstrates that during an in vitro neural correlate of eyeblink classical conditioning, there is a rapid and transient increase in levels of Arc protein that require activation of N-methyl-d-aspartate receptors. In the early phase of conditioning during conditioned response (CR) acquisition, there is significantly greater colocalization of Arc protein and GluR4-containing AMPA receptors at synaptic sites, however, colocalization of Arc and GluR4 was not observed after later stages of conditioning during CR expression. There was also significantly enhanced coimmunoprecipitation of Arc with GluR4 subunits and actin early in conditioning but not of Arc with NR1 subunits, and these associations declined to control levels in later stages of conditioning. These data suggest a role for Arc protein in the synaptic delivery of GluR4-containing AMPA receptors by interactions with cytoskeletal protein complexes during the acquisition phase of in vitro classical conditioning.

Animals↗

A cellular mechanism of classical conditioning in Aplysia.

The defensive siphon and gill withdrawal of Aplysia is a simple reflex, mediated by a well-defined neural circuit, that exhibits sensitization in response to strong stimulation of the tail. The siphon withdrawal reflex also exhibits classical conditioning when a weak stimulus to the siphon or mantle shelf (the conditioned stimulus or CS) is paired with a shock to the tail (the unconditioned stimulus or US). Cellular studies indicate that the mechanism of this conditioning shares aspects of the mechanism of sensitization of the reflex: presynaptic facilitation due to a cAMP-mediated decrease in K+ current and consequent broadening of action potentials in the sensory neurones. Thus, tail shock (the US) produces greater facilitation of the monosynaptic EPSP from a sensory to a motor neurone if the shock is immediately preceded by spike activity in the sensory neurone than if it occurs without spike activity (sensitization), or if the shock and spike activity are presented in a specifically unpaired pattern. This activity-dependent amplification of facilitation involves a greater broadening of action potentials in paired than in unpaired sensory neurones and appears to be due to a greater depression of the same serotonin- and cAMP-sensitive K+ current involved in sensitization. These results indicate that a mechanism of classical conditioning of the withdrawal reflex is an elaboration of the mechanism underlying sensitization. By analogy, the mechanisms of higher-order features of learning, such as the effect of contingency, may be built from combinations of the molecular mechanisms of these simple forms of learning.

Animals↗

Inactivation of the brachium conjunctivum prevents extinction of classically conditioned eyeblinks.

It is well established that the intermediate cerebellum is involved in the acquisition of classically conditioned eyeblink responses (CRs). Recent studies that inactivated the interposed nuclei (IN) demonstrated that blocking the intermediate cerebellum also interrupts CR extinction. Is this extinction deficit related to interrupting the information flow to efferent targets of the IN? To address this question, we inactivated axons of IN neurons in the brachium conjunctivum (BC). This treatment blocked the output of the intermediate cerebellum without directly affecting neurons in the deep cerebellar nuclei. Rabbits were trained in a delay classical conditioning paradigm, using a tone as the conditioned stimulus (CS) and a corneal air puff as the unconditioned stimulus (US). Then, the BC was microinjected with a sodium channel blocker, tetrodotoxin, during 4 extinction sessions in which rabbits were presented only with the CS. Tests performed after the 4-day injection period revealed that CRs did not extinguish in BC inactivation sessions but extinguished at a normal rate in the absence of the drug. CRs were then re-acquired. These data show that the normal flow of information along axons of cerebellar nuclear cells is required for CR extinction.

Animals↗