Classical conditioning of the rabbit eyelid response with a mossy-fiber stimulation CS: I. Pontine nuclei and middle cerebellar peduncle stimulation

The nictitating membrane/eyelid responses of 18 rabbits were classically conditioned using cerebellar mossy-fiber stimulation as a conditioned stimulus (CS) and air puff as an unconditioned stimulus (US). The dorsolateral, lateral, and medial pontine nuclei and the middle cerebellar peduncle were effective stimulation-CS sites for training. In one group of rabbits, robust conditioned eyelid responses were produced with paired trials and subsequently extinguished with CS-alone and explicitly unpaired presentation of the CS and US. In a second group of rabbits, no conditioned responses were evident for 4 days of unpaired CS and US presentations. Conditioned responses did develop, however, after paired training was begun. Lesions of the interpositus nucleus of the cerebellum completely abolished the conditioned responses of a third group of rabbits overtrained with the mossy-fiber CS and air-puff US. These results support previous studies which have demonstrated that the cerebellum is critically involved in acquisition and retention of simple learned responses. In addition, the present results support previous theories of cerebellar function which have proposed that mossy fibers supply critical "learning" input to the cerebellum for acquisition and retention of motor skills.     

Classical conditioning of the nictitating membrane response of the rabbit

The nictitating membrane response (NMR) of 20 rabbits was conditioned to light and white noise conditional stimuli (CSs) using a periorbital shock unconditional stimulus (US). Unilateral lesions of the cerebellar cortex, sparing the underlying deep nuclei, were then made. Small lesions of cerebellar cortical lobule HVI abolished conditioning on the side of the lesion to both CSs leaving unconditional responses to the US intact. Larger lesions of the posterior lobe which spared HVI did not impair NMR conditioning. We conclude that cerebellar lobule HVI is essential for NMR conditioning in the rabbit. Degeneration following critical lesions of HVI was seen in a restricted region of the inferior olive - the medial part of the dorsal accessory olive and the adjoining medial part of the dorsal leaf of the principal olive. This region of the olive provides somatosensory information from the face to HVI. We suggest that HVI receives information related to the US via climbing fibres from the olive and CS information via mossy fibres from the pontine nuclei. The critical changes underlying NMR conditioning may be the association of these two inputs at the Purkinje cells of cortical lobule HVI.     

Classical conditioning of the nictitating membrane response of the rabbit

Summary We report the connections of cerebellar cortical lobule HVI in the rabbit. We have studied the anterograde and retrograde transport of wheatgerm-agglutinated horseradish peroxidase (WGA-HRP) following its injection into HVI to reveal efferent and afferent connections. All of the cases showed strong anterograde transport to the anterior interpositus nucleus (AIP) — indicating that this is the major efferent target of HVI. Retrogradely labelled cells were found in the inferior olivary, spinal trigeminal, lateral reticular, inferior vestibular and pontine nuclei. Within the olive, the medial part of the rostral dorsal accessory olive (DAO) and the adjacent medial part of the principal olive (PO) were consistently labelled in all cases. This area is known to receive somatosensory information from the face and neck. There was no projection to the hemispheral part of lobule VI from visual parts of the olive within the dorsal cap and medial parts of the medial accessory olive. Likely sources of visual and auditory information to HVI are the dorsolateral basilar pontine nuclei and nucleus reticularis tegmenti pontis, which were densely labelled in all cases. These anatomical findings are consistent whith the suggestion that, during NMR conditioning, information related to the periorbital shock unconditional stimulus (US) may be provided by climbing fibres to HVI and light and white noise conditional stimulus (CS) information may be supplied by pontine mossy fibres.     

Classical conditioning of the nictitating membrane response of the rabbit

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.     

Classical conditioning of the nictitating membrane response of the rabbit

Summary The nictitating membrane response (NMR) of 15 rabbits was conditioned to light and white noise conditional stimuli (CSs) using a periorbital shock unconditional stimulus (US). Unilateral lesions of the inferior olive were then made. Lesions restricted to the medial parts of rostral dorsal accessory olive (DAO) and principal olive (PO) abolished conditioning and prevented subsequent acquisition on either side. Unconditional responses to the US were intact. Lesions in all other parts of the olive did not impair conditioning. The effective lesions were located in that part of the olive which supplies somatosensory information from the face to cerebellar lobule HVI. Lobule HVI is also essential for NMR conditioning. We suggest that this region of the inferior olive is part of a circuit which provides US information to the cerebellar cortex during NMR conditioning.     

Classical conditioning with brain stimulation at one site serving as both CS and US

Cats with chronic electrodes in the inferior colliculus received paired presentations of a weak (CS) and a strong (US) electrical stimulus delivered through the same electrode. After 140–200 such pairings, the weaker stimulus was able to elicit a close approximation to the unconditioned motor response elicited by the stronger stimulus. The response eventually elicited by the weaker stimulus was seen to extinguish following a series of presentations of the weaker stimulus alone. Additional animals receiving the same number of unpaired presentations of the weak and strong electrical stimuli failed to acquire the tendency to make a motor movement in the presence of the weak stimulus. The weak electrical stimulus coming to evoke a response originally elicited only by a stronger stimulus was explained on the basis of classical conditioning rather than pseudoconditioning or sensitization.     

Classical conditioning of a flexor nerve response in spinal cats: effects of tibial nerve CS and a differential conditioning paradigm

Previous studies have shown flexor nerve response increases produced by classical conditioning procedures in spinal cats when the conditioned stimulus (CS) was delivered to the superficial peroneal nerve and the unconditioned stimulus (US) was delivered to the ankle skin. In this study, these effects were produced when the CS was delivered to the whole tibial nerve or to the medial plantar branch. The finding that response increases followed by extinction effects could be obtained when either the superficial peroneal or the tibial nerve was utilized allowed the assessment of the effects of a differential conditioning paradigm. The responses to CS-US presentations on the superficial peroneal nerve increased, whereas responses to CS presentations on the tibial nerve remained unchanged. However, lack of extinction effects in the superficial peroneal data suggested that stimulation of the tibial nerve potentiated superficial peroneal evoked responses. Furthermore, responses evoked by stimulation of either nerve increased when paired trials were given on the tibial nerve. These data demonstrate that stimulation of the tibial nerve potentiates responses to superficial peroneal nerve stimulation but that superficial peroneal nerve stimulation has no effect on responses to CS presentations to the tibial nerve.     

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. © 1997 John Wiley & Sons, Inc. Dev Psychobiol 30: 49–59, 1997     

Classical conditioning of rabbits 2-1/2 to 4 years old using mossy fiber stimulation as a CS

A basic issue in age-related deterioration in memory formation concerns the locus of the age changes. Changes in peripheral portions of the auditory CS pathway could account for the observed age differences in classical conditioning in rabbits. We bypassed peripheral portions of the CS pathway and input the CS more centrally by electrical microstimulation of mossy fibers in the pons. New Zealand White rabbits aged 2-1/2-4 years were compared to six 3-month-old control rabbits with electrode implantations in comparable areas. Training consisted of 108 trials per day in which stimulation CS was paired with a corneal airpuff US. Older rabbits took almost five times as long to condition as the young animals. Thus, age differences in eyelid classical conditioning cannot be entirely attributed to changes in the peripheral CS pathway. Changes in the nervous system occurring more centrally in the cerebellum appear to affect learning and memory.     

Olfactory associative conditioning in infant rats with brain stimulation as reward: II. Norepinephrine mediates a specific component of the bulb response to reward.

One of the circuits modified by early olfactory learning is in the olfactory bulb. Specifically, response patterns of mitral-tufted cells are modified by associative conditioning during the early postnatal period. In addition, previous work has demonstrated that mitral-tufted cell single units respond to both olfactory conditioned stimuli and rewarding stimulation of the medial forebrain bundle-lateral hypothalamus (MFB-LH). The present study suggests that norepinephrine beta-receptor activation is required for early olfactory learning using MFB-LH stimulation as reward. Propranolol injected before odor-MFB-LH pairings blocks the acquisition of conditioned behavioral responses and their neural correlates to the conditioned odor. Furthermore, propranolol blocks a specific class of the mitral-tufted cell responses to MFB-LH reward stimulation. The relationship of this response to reward and early learning is discussed.     

Electrical stimulation of brainstem nuclei: elicitation, modification, and conditioning of the rabbit nictitating membrane response

Electrical stimulation of the reticular formation, pars oralis of the spinal trigeminal, abducens, and accessory abducens nuclei was used to assess the role of these sites in the elicitation, reflex modification, and classical conditioning of the rabbit's nictitating membrane response (NMR). Although electrical brain stimulation of the targeted sites revealed comparable levels of unconditioned responses, the spinal trigeminal nucleus was the only site at which reflex modification and conditioned response acquisition occurred reliably. These findings suggest that a locus of conditioned stimulus and unconditioned stimulus interaction, mediating either or both reflex modification and NMR conditioning, is on the sensory side of the reflex arc, at the pars oralis of the spinal trigeminal nucleus.     

Amygdala central nucleus lesions: effect on heart rate conditioning in the rabbit

The present study was designed as an initial step in determining the specific anatomical systems of the amygdala which may contribute to the expression of conditioned heart rate responding during aversive Pavlovian conditioning in the New Zealand rabbit. Animals receiving either small or large radio-frequency lesions of the central nucleus of the amygdala demonstrated a significant attenuation of the conditioned bradycardia response to the conditioned stimulus when compared to that demonstrated by control animals. No significant effects of the lesions on baseline heart rate or on the heart rate orienting response to the conditioned stimulus were observed. Central nucleus lesions also produced a significant effect on the unconditioned heart rate response to the unconditioned stimulus manifested in an increased duration and prolonged habituation of the unconditioned response. The results are consistent with anatomical and physiological evidence suggesting the involvement of the central nucleus in the expression of emotional responses including accompanying cardiovascular alterations.     

The lateral reticular nucleus in the cat—II. Effects of lateral reticular lesions on posture and reflex movements

Unilateral lesion of the lateral reticular nucleus produced a postural asymmetry characterized by ipsilateral hypertonia and contralateral hypotonia of the limb extensor muscles. Soon after the operation the cat was unable to stand or walk, and it laid on one side. Within 3 days, it began to walk, but it often deviated to its contralateral side, 'frequently falling in this direction. Some compensation for both postural and motor deficits occurred in chronic preparations maintained up to 154 days after the lesion. The postural asymmetry was reversed by the following operations: (1) section of the ipsilateral VIIIth nerve, (2) electrolytic lesion of the ipsilateral Deiters' nucleus, or (3) ablation of the contralateral vermal cortex of the cerebellar anterior lobe.The lateral reticular nucleus lesion also produced, in the ipsilateral limbs, a transient loss of the proprioceptive placing reaction and a persistent deficit of the tactile placing reflex. These effects on the ipsilateral side were not reversed by the procedures described above.All of these behaviors depended on selective destruction of the lateral reticular nucleus and were not due to damage to the nearby main reticular formation or ascending and descending pathways. Moreover, the postural changes did not involve mesencephalic or higher mechanisms, since they were still observed after decerebration. The forelimbs were affected primarily by lesions involving the dorsomedial, magnocellular part of the lateral reticular nucleus, whereas the hindlimbs also were affected by lesions including the ventrolateral, parvicellular part of the lateral reticular nucleus.The postural asymmetry is attributed to interruption of the crossed spinoreticulocerebellar pathway, acting on the vermal cortex of the anterior lobe and the fastigial nucleus, while the ipsilateral loss of the placing reaction is attributed to interruption of the uncrossed spinoreticulocerebellar pathway acting on the intermediate cortex of the anterior lobe and the interpositus nucleus. The lateral reticular nucleus appears, therefore, to be composed of two, more or less independent, parts, a crossed one being related to major changes in postural tone, equilibrium and locomotion of the entire body, and an uncrossed one restricted to discrete movements of the ipsilateral limbs.     

Eyeblink conditioning in the rabbit (Oryctolagus cuniculus) with stimulation of the mystacial vibrissae as a conditioned stimulus.

Eyeblink conditioning is a well-understood paradigm for the study of learning and memory and has been successfully employed with the use of auditory and visual conditioned stimuli (CSs). In this study, vibrotactile stimulation of the mystacial vibrissae was examined as an alternative CS in the rabbit (Oryctolagus cuniculus). The technique is described and acquisition of eyeblink conditioning (EBC) with stimulation of a single row of vibrissae in a delay paradigm is reported. Extinction of EBC with presentation of the CS alone is demonstrated, as well as reacquisition with stimulation of a single whisker. Finally, control experiments ensure that the CS has no auditory components. Ipsilateral presentation of the CS and airpuff is a more effective combination for training than contralateral presentations. Vibrotactile stimulation of the vibrissae as a CS will enable further examination of the neural correlates of learning in a well-characterized sensory system.     

Differential response dynamics of corticothalamic glutamatergic synapses in the lateral geniculate nucleus and thalamic reticular nucleus

The corticothalamic feedback pathway provides excitatory synaptic input to both the thalamic reticular nucleus and the lateral geniculate nucleus. We studied excitatory postsynaptic currents elicited from corticothalamic stimulation in the visual sector of the thalamic reticular nucleus and the lateral geniculate nucleus to compare the response of these neurons to stimulation of their common input pathway. Using whole cell patch clamp recordings in ferret thalamic slices, we compared single excitatory postsynaptic current decay kinetics, presynaptic glutamate release dynamics through paired pulse facilitation and responses to corticothalamic train stimulation. We found that single thalamic reticular nucleus excitatory postsynaptic currents were significantly sharper than lateral geniculate nucleus responses. The mean thalamic reticular nucleus excitatory postsynaptic current decay constant (tau) was 4.9+/-0.5 ms, while the mean lateral geniculate nucleus excitatory postsynaptic current tau value was 11.8+/-0.8 ms. Presynaptic release dynamics as measured by responses to paired stimuli were conserved between the thalamic reticular nucleus and lateral geniculate nucleus. However, facilitating responses to train stimulation were markedly different between nuclei. Lateral geniculate nucleus responses showed proportionately larger facilitation (reaching 842.9 +/- 76.4% of excitatory postsynaptic current 1 amplitude) than thalamic reticular nucleus responses (reaching 223.1 +/- 44.0% of excitatory postsynaptic current 1 amplitude). These data indicate that while the corticothalamic pathway produces excitatory postsynaptic currents in both the thalamic reticular nucleus and lateral geniculate nucleus, other factors uniquely affect the functional integration of the inputs in each nucleus.     

Retardation of rabbit nictitating membrane conditioning by subseizure electrical stimulation of hippocampus.

The experiment involved classical conditioning of the nictitating membrane response of rabbits with subseizure, posttrial electrical stimulation of hippocampus. Hippocampal electrical activity was recorded before, during and after each trial to assure that no seizure activity was present. Stimulated animals, in comparison with unstimulated controls, showed a marked retardation of CR onset which paralleled that of previous research employing seizure producing stimulation. This implies that brain seizures are not the critical element in the disruption effect, but rather the anatomical locus of stimulation. Results are discussed in the context of recent lesion, recording and stimulation research employing the conditioned nictitating membrane response of rabbits as a model system.     

Classical conditioning of heart rate in rats using direct vagal stimulation as a US

Classically conditioned heart-rate deceleration was established in rats using a 2-sec stimulation of the intact cervical vagus nerve as the US. Although small in magnitude, changes in the form of the CR over training indicated that there may have been inhibition of delay similar to that obtained using a traditional painful shock US. Independent assessments of motivational potency revealed that the vagal US did not lead to conditioned lever-press suppression in a CER procedure, but did produce reliable escape responding in a shuttle-box preference situation. It was suggested that the mildly aversive motivational effects of vagal stimulation may have stemmed from the activation of a vagally mediated system involved in the regulation of hunger.