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In vitro classical conditioning of a gill withdrawal reflex in Aplysia: neural correlates and possible neural mechanisms.

An in vitro preparation consisting of the siphon, mantle, gill, and abdominal ganglion undergoes classical conditioning when a weak tactile stimulus (CS) applied to the siphon is paired with a strong tactile stimulus to the gill (UCS). When the stimuli are paired, the CS comes to evoke a gill withdrawal reflex (GWR) which increases in amplitude with training. Only when the stimuli are paired in a classical conditioning paradigm does the CS come to evoke a GWR. With classical conditioning training there is an alteration in the synaptic efficacy between central sensory neurons and central gill motor neurons. Moreover, these changes can be observed in sensory neurons not activated by the CS. The changes observed, as evidence by the number of action potentials evoked in the gill motor neuron do not completely parallel the observed behavioral changes. It is suggested that in addition to changes in the synaptic efficacy at the sensory-motor neuron synapse, other changes in neuronal activity occur at other loci which lead to the observed behavioral changes.

Animals↗

Classical conditioning of the nictitating membrane response of the rabbit. I. Lesions of the cerebellar nuclei.

The classically conditioned nictitating membrane response (NMR) of the rabbit, a simple form of associative motor learning, is crucially dependent upon the cerebellum. Discrete unilateral lesions of the cerebellar nuclei were made in 20 rabbits. Lesions of the anterior interpositus nucleus (IA) abolished NMR conditioning to light and white noise stimuli on the side of the lesion without affecting unconditional responses. Lesions of the posterior interpositus nucleus, fastigial and dentate nuclei were without effect upon NMR conditioning.

Animals↗

Nicotinic cholinergic system involvement in eyeblink classical conditioning in rabbits.

A nicotinic cholinergic antagonist, mecamylamine (MEC), was administered to rabbits tested on eyeblink classical conditioning (EBCC) in the 750-ms delay paradigm for 10 90-trial sessions. Nicotinic receptors were measured in 3 brain regions in 5 treatment groups: paired conditioned stimulus-unconditioned stimulus (CS-US) presentations with (a) vehicle, young; (b) vehicle, older; (c) 0.5 mg/kg MEC, young; unpaired CS-US with (d) 0.5 mg/kg MEC, young; and (e) vehicle, young. Daily MEC injections disrupted acquisition in young rabbits (769 trials to learning criterion vs. 323 trials for vehicle-treated young rabbits). MEC-treated young rabbits learned similarly to older rabbits. Brain nicotinic receptors were not affected by 10 daily MEC injections. To our knowledge, this experimental protocol, using a low MEC dose to selectively inhibit nicotinic cholinergic receptors, is the first to demonstrate a role for nicotinic cholinergic receptors in EBCC.

Animals↗

Exploratory studies of classical conditioning of the preoral cavity in harnessed carpenter ants (Camponotus pennsylvanicus).

An attempt was made to classically condition the mouthparts of harnessed worker ants (Camponotus pennsylvanicus) in anticipation of feeding. Experiments were designed to investigate classical conditioning with one CS, discrimination between two CSs, and pseudoconditioning. Analysis indicated a small acquisition effect that could be accounted for by pseudoconditioning. The preparation can be used to study nonassociative learning and some instrumental conditioning situations.

Animals↗

Classical conditioning in the fetal rat: reinforcing properties of dynorphin A (1-13).

This study examined the reinforcing properties of dynorphin A (1-13) in a single-trial classical conditioning paradigm in the E20 rat fetus. Injection of dynorphin into the cisterna magna increased fetal motor activity and reduced facial wiping in a test of perioral cutaneous responsiveness. Dynorphin was effective as an unconditioned stimulus (US) in a classical conditioning paradigm using an artificial nipple conditioned stimulus (CS) and dynorphin A (1-13) US. The association between CS and US was dependent on activity in the kappa opioid system. Re-exposure to the artificial nipple CS after a single pairing of the nipple with dynorphin resulted in conditioned activation of the kappa opioid system. Dynorphin A (1-13) functions as a reinforcer for classical conditioning in the rat fetus after intracisternal or intrahemispheric injection, with the conditioned response depending on route of administration and site of injection.

Animals↗

Acquisition of classically conditioned eyeblink response following bilateral lesions of flocculus and paraflocculus.

Stimulation of regions of the cerebellar flocculus can elicit eyeblinks, but the relationship of this floccular eyeblink zone to eyeblink classical conditioning is unknown. In this experiment, New Zealand white rabbits received bilateral lesions of the flocculus and paraflocculus and were subsequently classically conditioned with tone and corneal airpuff on the left and then the right eye. All animals reached training criterion on both eyes, with the exception of one animal whose lesion included the superior cerebellar peduncle and who was unable to learn on the ipsilateral eye. The lesioned group was not significantly different from unlesioned controls in rate of acquisition or conditioned or unconditioned response amplitude. These results indicate that the flocculus and paraflocculus are not by themselves the essential site of plasticity for classical conditioning of the rabbit eyeblink response.

Animals↗

Age-related disruption of trace but not delay classical conditioning of the rabbit's nictitating membrane response.

Young (6 months of age) and old (36-60 months) New Zealand albino rabbits underwent classical conditioning of the nictitating membrane response in either a delay conditioning (Experiment 1) or a trace conditioning (Experiment 2) paradigm. There was no difference between old and young animals in acquisition of the conditioned response in the delay paradigm, nor were there any age-related differences in generalization to the tone conditioned stimulus (CS) or in sensitivity to the tone CS or eye shock unconditioned stimulus. In the trace conditioning paradigm, however, old animals acquired the conditioned response significantly more slowly than young rabbits. Because the same stimulus parameters and the same response were used in both experiments, it is unlikely that age-related differences in trace conditioning were due to stimulus sensitivity, motivation, or fatigue. The results are discussed in terms of how brain changes that accompany aging could differentially affect these two types of classical conditioning.

Aging↗

Classical conditioning of beginning reading responses.

The present study describes the first demonstration that laboratory-controlled experimental procedures can lead to the successful acquisition and subsequent retention of classically conditioned beginning reading responses (CCBRRs) in children of both sexes and mean age of 4 yr. Anticipatory instructions combined with higher-order classical conditioning temporally arranged into a trace conditioning paradigm presented for 10 trials for each response to be learned led to beginning reading responses being successfully acquired by 20 children during 95% of the 2,220 total acquisition learning trials and subsequently correctly recalled on 114 of the 222 retention test trials. Findings support the view that perhaps the relatively sudden and sustained acquisition learning curves for reading responses on the second-signalling-system level of behavior in the present study may be quite different from the relatively slow and incremental learning curves usually obtained in classical conditioning of the autonomic type which occur on the first-signalling-system level.

Age Factors↗

Sensory and associative effects of morphine and naloxone in classical conditioning of the rabbit nictitating membrane response.

In Experiment I, classical conditioning of the rabbit's nictitating membrane response was accomplished by the pairing of tone and light conditioned stimuli with a shock unconditioned stimulus applied to the paraorbital region of the head. Morphine (5 mg/kg) significantly retarded the acquisition of conditioned responses to both conditioned stimuli. Moreover, morphine had no effect on nonassociative responding (baseline responding or responding to tone and light stimuli) or on the latency and amplitude of the unconditioned response elicited by shock during the explicitly unpaired presentations of tone, light and shock stimuli. The retardant effect of morphine on acquisition of conditioned responses was blocked by naloxone (1 mg/kg). In Experiment II, morphine (0.2-10 mg/kg) had no effect on the intensity threshold of the shock unconditioned stimulus for elicitation of unconditioned responses or on the latencies of the elicited responses. However, morphine (5 and 10 mg/kg) did produce a small but significant decrease in the amplitude of unconditioned responses elicited by the two highest shock intensities employed (3 and 4 mA). This latter effect of morphine was completely blocked by naloxone (1 mg/kg). In Experiment III, morphine (5 mg/kg) blocked the sensory processing of a tone conditioned stimulus, in previously trained animals, as measured by a significant (24 dB) elevation in the intensity threshold of the conditioned stimulus for elicitation of conditioned responses and an increase in the latency of the elicited response. Naloxone (1 mg/kg) completely blocked the effects of morphine on the sensory processing of the tone-conditioned stimulus.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Classical conditioning in the sea anemone, Cribrina xanthogrammica.

Earlier attempts to classically condition sea anemones have yielded inconclusive results. Using light as CS and shock as US, conditioning as distinguished from sensitization and pseudoconditioning was demonstrated in anemones. Procedural controls included substitution of light alone, shock alone, and random light and shock in place of paired light-shock trials. Responses measured were electrical output and folding of the oral disc. The conditioned response was distinguished from the unconditioned response to light and the unconditioned response to shock in terms of response latency of both electrical and behavioral measures.

Cnidaria↗

Classically conditioned changes of blood glucose level in humans.

Procedures of classical conditioning in animals and man have provided evidence that most psychophysiological responses can be acquired by repeated association with previous neutral stimuli. Many animal studies reported on conditioned changes in the blood glucose level; the nature of the conditioned response (CR), hypo- or hyperglycemia, however, seems to vary with experimental procedures. The present study aimed to elicit conditioned blood glucose changes in human subjects. Thirty male volunteers participated in five sessions each. The sessions were separated by 3 days, with identical time course and procedure. The subjects were informed that we wanted to test the effects of insulin or placebo injections on cognitive functioning, and were kept busy with pseudotests. In four sessions, subjects were injected with 0.035 IU/kg body weight of human insulin as the unconditioned stimulus (US), which induced the expected fall in blood glucose level below 50 mg/dl (UR). Injections were accompanied by a specific stimulus compound (conditioned stimulus, CS) in half of the subjects. In the fifth session, the CS was associated with a placebo injection. About half of the subjects showed a change from the baseline level of blood glucose of more than 10 mg/dl, which we would interpret as a conditioned response. Conditioning occurred more often in those subjects who were given a CS compound in addition to the injection, which itself together with the experimental environment may have been a sufficient CS.

Adult↗

Effects of MDA on classical conditioning of the rabbit nictitating membrane response.

In Experiment 1, classical conditioning of the rabbit's nictitating membrane response (NMR) was accomplished by pairing tone and light conditioned stimuli (CSs) with a shock unconditioned stimulus (UCS). MDA impaired the acquisition of conditioned responses (CR) to a tone-CS, while significantly enhancing CR acquisition to a light-CS. Experiment 2, employing explicitly unpaired CS, UCS training, revealed no reliable effects of MDA upon nonassociative processes. Subsequent efforts determined if MDA's CR acquisition effects resulted from alterations in sensory processing of the CS, UCS, and/or UCR motor functioning. Specifically, it was determined that MDA: (a) increased the tone-CS intensity threshold for eliciting CRs (Experiment 3); (b) attenuated the tone-induced reflex modification of the unconditioned NMR (Experiment 4); and (c) enhanced UCR frequency at varying UCS intensities (Experiment 5). It was concluded that MDA's effect upon CR acquisition reflected the drug's effect upon CS and UCS/UCR processing and thereby altered the ability of these components of conditioning to enter into associative learning.

3,4-Methylenedioxyamphetamine↗

Scopolamine disruption of behavioral and hippocampal responses in appetitive trace classical conditioning.

Twelve young rabbits (3-6 months; Oryctolagus cuniculus) were classically conditioned in a trace jaw movement paradigm (300 ms tone, 450 ms trace, 200 ms intraoral water) after implantation of electrodes into area CA1 of dorsal hippocampus. Rabbits were divided into two groups and administered either 0.5 mg/kg scopolamine hydrobromide (HBr) or 0.5 mg/kg scopolamine methylbromide (MBr) subcutaneously before daily training sessions. Rabbits given HBr took significantly more trials to reach a behavioral criterion of eight conditioned responses in any nine consecutive trials than rabbits given MBr (P = 0.03). Conditioned, but not unconditioned, rhythmic jaw movement responses of the HBr group were of a lower frequency (Hz) than those of MBr rabbits (P = 0.02). The magnitude of hippocampal conditioning-related responses across the first 3 days of training was significantly smaller for HBr rabbits than for MBr rabbits (P = 0.02). These effects of central cholinergic blockade are similar to those reported for undrugged aging rabbits trained in the same paradigm (Seager MA, Borgnis RL, Berry SD. Neurobiol. Aging 1997;18(6):631 639).

Age Factors↗

The effects of reversible inactivation of the red nucleus on learning-related and auditory-evoked unit activity in the pontine nuclei of classically conditioned rabbits.

The pontine nuclei carry auditory conditioned stimulus information to the cerebellum during classical conditioning of the nictitating membrane response in rabbits. In well-trained animals learning-related as well as stimulus-evoked unit activity can be recorded throughout the pontine nuclei but particularly in the lateral and dorsolateral pons. Recent work in our laboratory has provided evidence that the learning-related unit activity in the pons is dependent on the interpositus nucleus and that the pons is not a site of essential plasticity for the learned response. In the present study we considered the question of whether learning-related unit activity might be projected from the interpositus nucleus to the pons through the red nucleus, a primary output target of the interpositus and a structure known to be essential for expression of the learned response. Multiple unit recordings were taken from lateral and dorsolateral pontine locations in well-trained rabbits before and during cooling of the red nucleus. Analysis of pooled data for all recording locations within the lateral and dorsolateral pons indicated that reversible inactivation of red nucleus abolished both stimulus-evoked and learning-related unit activity. However, we also found discrete recording locations where stimulus-evoked and learning-related unit activity were attenuated but not abolished by red nucleus cooling.

Animals↗

[Classical conditioning of anaphylaxis in sensitized guinea pigs].

An attempt of classical conditioning of anaphylaxis by odor was carried out, using actively sensitized guinea pigs with an inhalation of ovalbumin (OA). One month after sensitization, animals were divided into the conditioned group; group C, and the unconditioned group; group U, consisted of 6 animals, respectively. Dimethylsulfied (DMS: sulfur odor), was inhaled in group C as a conditioned stimulus with OA which is an unconditioned stimulus, while only OA was inhaled in group U. Four days after these procedures, saline was inhaled in group C and DMS was solely inhaled in group U in order to equalize the total inhaled dose of OA and DMS in both groups. These sessions were repeated once a week for seven weeks. After final sessions, all animals were inhaled DMS, saline and OA separately, and blood samples were drawn after each inhalation to measure plasma histamine levels. After an inhalation of DMS only, plasma histamine levels of group C and U were 47.5 +/- 9.7 and 25.7 +/- 1.2 ng/ml, respectively. In group C, plasma histamine levels were 32.9 +/- 4.7 at the inhalation of saline and 59.0 +/- 9.2 ng/ml at OA inhalation. Plasma histamine level after an inhalation of DMS only was significantly higher in group C than that in group U (p < 0.05). These results suggest that the conditional stimulus (DMS inhalation) may induce histamine release in the absence of any antigenic stimulus and support the evidence for mast cell-neuron interaction.

Anaphylaxis↗

Effects of LSD on learning as measured by classical conditioning of the rabbit nictitating membrane response.

Acquisition of the classically conditioned, nictitating membrane response was used to assess effects of LSD on learning. Tone and light conditioned stimuli (CS) were presented 800 msec before delivery of the unconditioned stimulus, consisting of a 100 msec electric shock to the skin over the paraorbital region of the head. Extension of the membrane to the CSs in the 800 msec prior to shock onset was recorded as a conditioned response (CR), while extension to shock onset was recorded as an unconditioned response (UCR). LSD (1, 10, 30, 100 or 300 nmol/kg) was injected i.v., 30 min before each daily conditioning session. Dosages of 1 to 100 nmol/kg of LSD produced a dose-dependent enhancement of CR acquisition. Acquisition to a criterion of 10 successive CRs required 184 trials at 30 nmol/kg of LSD as compared to 293 trials with controls. Separate groups of rabbits received explicity unpaired presentations of stimuli (tone alone, light alone and shock alone). The frequency of responding within 800 msec of CS onset or in the 800 msec before shock onset was low (2-3%) and was not affected by any dosage of LSD, indicating that the effects of LSD on acquisition were not due to sensitization, pseudoconditioning or changes in baseline responding. LSD also had no effect on UCR amplitude. Hence, the systematic effects of LSD on acquisition of CRs reflects the action of the drug on learning.

Acoustic Stimulation↗

Intracellular correlates of acquisition and long-term memory of classical conditioning in Purkinje cell dendrites in slices of rabbit cerebellar lobule HVI.

Intradendritic recordings in Purkinje cells from a defined area in parasaggital slices of cerebellar lobule HVI, obtained after rabbits were given either paired (classical conditioning) or explicitly unpaired (control) presentations of tone and periorbital electrical stimulation, were used to assess the nature and duration of conditioning-specific changes in Purkinje cell dendritic membrane excitability. We found a strong relationship between the level of conditioning and Purkinje cell dendritic membrane excitability after initial acquisition of the conditioned response. Moreover, conditioning-specific increases in Purkinje cell excitability were still present 1 month after classical conditioning. Although dendritically recorded membrane potential, input resistance, and amplitude of somatic and dendritic spikes were not different in cells from paired or control animals, the size of a potassium channel-mediated transient hyperpolarization was significantly smaller in cells from animals that received classical conditioning. In slices of lobule HVI obtained from naive rabbits, the conditioning-related increases in membrane excitability could be mimicked by application of potassium channel antagonist tetraethylammonium chloride, iberiotoxin, or 4-aminopyridine. However, only 4-aminopyridine was able to reduce the transient hyperpolarization. The pharmacological data suggest a role for potassium channels and, possibly, channels mediating an IA-like current, in learning-specific changes in membrane excitability. The conditioning-specific increase in Purkinje cell dendritic excitability produces an afterhyperpolarization, which is hypothesized to release the cerebellar deep nuclei from inhibition, allowing conditioned responses to be elicited via the red nucleus and accessory abducens motorneurons.

Action Potentials↗

Acquisition of the classically conditioned eyeblink response in humans over the life span.

Human subjects ranging in age from 18 to 85 years underwent classical conditioning of the eyeblink response to a tone conditioned stimulus (CS) and an air-puff unconditioned stimulus (UCS). There was a decline in percentage of conditioned responses with age. This decline was most noticeable in subjects over age 50. These conditioning deficits were not due to age-related changes in sensitivity to the tone CS or the air-puff UCS, nor could the conditioning deficits be attributed to an age-related decline in general cognitive abilities or to changes in spontaneous blink rates. The results are discussed in terms of using the classically conditioned eyeblink in humans in conjunction with the classically conditioned nictitating membrane response in rabbits as a model system for studying the neurobiology of age-related conditioning deficits.

Adolescent↗