Inactivation of interposed nuclei in the cat: classically conditioned withdrawal reflexes, voluntary limb movements and the action primitive hypothesis

 The cerebellar interposed nuclei are considered critical components of circuits controlling the classical conditioning of eyeblink responses in several mammalian species. The main purpose of the present experiments was to examine whether the interposed nuclei are also involved in the control of classically conditioned withdrawal responses in other skeletomuscular effector systems. To achieve this objective, a unique learning paradigm was developed to examine classically conditioned withdrawal responses in three effector systems (the eyelid, forelimb and hindlimb) in individual cats. Trained animals were injected with muscimol in the cerebellar interposed nuclei, and the effects on the three conditioned responses (CRs) were examined. Although the effects of muscimol were less dramatic than previously reported in the rabbit eyeblink preparation, the inactivation of the cerebellar nuclei affected the performance of CRs in all three effector systems. In additional experiments, animals were injected with muscimol at the sites affecting classically conditioned withdrawal responses to determine the effects of these injections on reaching and locomotion behaviors. These tests demonstrated that the same regions of the cerebellar interposed nuclei which control withdrawal reflexes are also involved in the control of limb flexion and precision placement of the paw during both locomotion and reaching tasks. The obtained data indicate that the interposed nuclei are involved in the control of ipsilateral action primitives and that inactivating the interposed nuclei affects several modes of action of these functional units. Received: 15 June 1998 / Accepted: 5 November 1998     

Intermediate cerebellum and conditioned eyeblinks

The intermediate cerebellum (the intermediate cerebellar cortex and interposed nuclei) and associated brainstem circuits are essential for the acquisition and expression of classically conditioned eyeblinks in the rabbit. The purpose of the present experiment was to determine whether these circuits are also involved in adaptive eyelid closure learned in an instrumental paradigm. For that purpose, rabbits with unrestrained eyelids were trained in two tasks: (1) classical conditioning of the eyeblink; and (2) a new instrumental task in which they avoided delivery of an aversive stimulus by maintaining tonic eyelid closure. To examine the involvement of the intermediate cerebellum in these two types of learned behavior, the cerebellar interposed nuclei were injected with the GABAA agonist muscimol and with the GABAA antagonist picrotoxin. Inactivating the interposed nuclei with muscimol abolished classically conditioned eyeblinks and severely affected the rabbit's capacity to maintain tonic eyelid closure. On the other hand, reducing inhibition with picrotoxin failed to interrupt the learned responses and increased the amplitude of eyelid closure. These data indicate that the cerebellar interposed nuclei control both phasic classically conditioned eyeblinks and tonic instrumental eyelid closure. To account for this new finding, a "hybrid" hypothesis combining the cerebellar learning hypothesis and the performance hypothesis is proposed.     

Effects of muscimol inactivation of the cerebellar interposed-dentate nuclear complex on the performance of the nictitating membrane response in the rabbit

Intracranial microinjections of the GABA_A agonist muscimol were used to assess the involvement of the dentato-interposed cerebellar nuclear complex in the performance of the conditioned (CR) and unconditioned (UR) nictitating membrane responses in the rabbit. Specifically, the experiments test the hypothesis that the cerebellar nuclei are involved in the performance of both the CRs and URs. The experiments employed temporary nuclear lesions to disrupt the CRs in order to examine parallel effects on URs. Animals were conditioned in a standard delay conditioning paradigm. Injection sites at which the muscimol application disrupted execution of the CRs were identified in each rabbit. Once these sites were found, the effects of muscimol and saline injections were evaluated while alternating paired trials with unpaired trials in which only the unconditioned stimuli were applied. There are two main findings in the present study. First, the activation of the GABA_A receptors in the dentato-interposed cerebellar nuclear region reduced the amplitude and increased the latency of the UR. This change in the UR closely paralleled the disruption of the CR. This observation is consistent with the notion that the cerebellum is involved in the regulation of defensive flexion reflexes. Second, cerebellar nuclear inactivation did not eliminate the tone-induced enhancement of the UR. This finding suggests the presence of cerebellum-independent circuits subserving the intermodal interaction between the conditioned and unconditioned stimuli.     

Effects of muscimol inactivation of the cerebellar interposed-dentate nuclear complex on the performance of the nictitating membrane response in the rabbit

Intracranial microinjections of the GABA_A agonist muscimol were used to assess the involvement of the dentato-interposed cerebellar nuclear complex in the performance of the conditioned (CR) and unconditioned (UR) nictitating membrane responses in the rabbit. Specifically, the experiments test the hypothesis that the cerebellar nuclei are involved in the performance of both the CRs and URs. The experiments employed temporary nuclear lesions to disrupt the CRs in order to examine parallel effects on URs. Animals were conditioned in a standard delay conditioning paradigm. Injection sites at which the muscimol application disrupted execution of the CRs were identified in each rabbit. Once these sites were found, the effects of muscimol and saline injections were evaluated while alternating paired trials with unpaired trials in which only the unconditioned stimuli were applied. There are two main findings in the present study. First, the activation of the GABA_A receptors in the dentato-interposed cerebellar nuclear region reduced the amplitude and increased the latency of the UR. This change in the UR closely paralleled the disruption of the CR. This observation is consistent with the notion that the cerebellum is involved in the regulation of defensive flexion reflexes. Second, cerebellar nuclear inactivation did not eliminate the tone-induced enhancement of the UR. This finding suggests the presence of cerebellum-independent circuits subserving the intermodal interaction between the conditioned and unconditioned stimuli.     

Reversible inactivations of the cerebellum with muscimol prevent the acquisition and extinction of conditioned nictitating membrane responses in the rabbit

Lesions of the cerebellum severely impair the classically conditioned nictitating membrane response (NMR) in rabbits. Thus, the cerebellum is essential for the production of conditioned responses (CRs), either because it is actively involved in NMR conditioning or because damage to it causes motor or other general deficits. To distinguish between these alternatives, the cerebellum may be inactivated during training. Inactivation of the cerebellum during acquisition training might result in the absence of CRs on initial trials of subsequent training without the neuronal blockade. The blockade may have prevented learning but it may have produced other deficits that require time or further training to overcome. This problem can be addressed by inactivating the cerebellum during extinction training. If inactivation during extinction training results in the immediate production of CRs when training is resumed without the blockade, then it may be concluded that extinction learning was prevented by the blockade — the presence of CRs argues against any deficits not associated with learning. We used muscimol to inactivate the cerebellum and test its involvement in acquisition and extinction of NMR conditioning in the same subjects. We injected muscimol close to the interpositus nucleus of the cerebellum 1 h before each of four daily training sessions of delay conditioning. Almost no CRs were produced in these training sessions — there was little or no acquisition of NMR conditioning during cerebellar inactivation. The subjects were then trained for four daily sessions without injections of muscimol. There were no CRs on initial trials of the first session of retraining, but all subjects produced CRs by the end of this session. The subjects then received four daily sessions of extinction training with muscimol inactivation of the nuclei — no CRs were produced. Extinction training then continued for four daily sessions without muscimol inactivation. On the first of these sessions, all subjects immediately produced high levels of CRs. These responses then extinguished within and between sessions with characteristic beginning-of-session spontaneous recovery. There was little or no extinction of NMR conditioning during cerebellar inactivation. After inactivation, the muscimolinactivated subjects went on to acquire and extinguish NM responses at rates similar to those of appropriate controls. We conclude that cerebellar circuitry is essential for, and actively engaged in, both acquisition and extinction of this simple form of motor learning.     

Functional plasticity in the interposito-thalamo-cortical pathway during conditioning

In classic conditioning, the interstimulus interval (ISI) between the conditioned (CS) and unconditioned (US) stimulus is a critical parameter. The aim of the present experiment was to assess whether, during conditioning, modification of the CS-US interval could reliably produce changes in the functional properties of the interposito-thalamo-cortical pathways (INTCps). Five cats were prepared for chronic stimulation and recording from several brain regions along this pathway in awake animals. The CS was a weak electric shock applied on the interposed nucleus of the cerebellum in sites that initially elicited forelimb flexion (i.e., alpha motor responses) in three cats, and equal proportions of flexor and extensor responses in two cats. The US was an electric shock applied on the skin that elicited forelimb flexions. The motor and neurobiological effects of synchronous CS-US were compared with pairings in which the CS was applied 100 ms before US. Simultaneous and sequential application of CS and US produced different behavioral outcomes and resulted in different neural processes in the interposito-thalamo-cortical pathways (INTCps). The simultaneous presentation of stimuli only produced a small increase in excitability spreading to all the body representational zones of the primary motor cortex and a weak increase in the amplitude of the alpha motor response. In contrast, the sequential application led to a profound modification of the interposed output to neurons in the forelimb representation of the motor cortex. These robust neuronal correlates of conditioning were accompanied by a large facilitation of the alpha motor response (alpha-MR). There were also changes in the direction of misdirected alpha responses and an emergence of functionally appropriate, long-latency withdrawal forelimb flexion. These data revealed that, during conditioning, plastic changes within the thalamocortical connections are selectively induced by sequential information from central and peripheral afferents. This sequence significantly contributed to neural processes that are responsible for the acquisition, expression, and extinction of anticipatory flexion responses.     

The temporary inactivation of the red nucleus affects performance of both conditioned and unconditioned nictitating membrane responses in the rabbit

These experiments are part of a series of studies examining the role of the red nucleus in the performance of the conditioned and unconditioned nictitating membrane reflexes in the rabbit. Specifically, the experiments test the hypothesis that the temporary inactivation of the red nucleus selectively affects the performance of the conditioned reflex. The experiments were designed to assess the effects of lidocaine and control saline microinjections on conditioned as well as unconditioned responses in both paired and unpaired trials. Rabbits were chronically implanted with cannulae through which small injecting tubes were passed stereotaxically to the red nucleus. The animals were conditioned using a delay paradigm in which a 1 kHz tone and an air puff applied to the cornea were used as the unconditioned and conditioned stimulus, respectively. Once conditioned, the effects of either lidocaine or saline injection were evaluated while alternating paired trials with unpaired trials in which only the air puff was applied. The principal finding of this study was that the amplitudes of both the conditioned and unconditioned responses were reduced following lidocaine injection into the red nucleus. The effect on the unconditioned response amplitude could not be ascribed to any interaction between the conditioned and unconditioned responses, since it also was present in the unpaired trials. The reduction in amplitude of the conditioned and unconditioned responses was shown to be correlated with changes in other characteristics of the same responses. The data suggest that the red nucleus contributes to the performance of both the conditioned and unconditioned nictitating membrane reflexes and consequently is not likely to be involved only in pathways responsible for mediating and/or storing the engram for the conditioned reflex.     

GABA neurotransmission in the cerebellar interposed nuclei: involvement in classically conditioned eyeblinks and neuronal activity

The cerebellar interposed nuclei (IN) are an essential part of circuits that control classically conditioned eyeblinks in the rabbit. The function of the IN is under the control of GABAergic projections from Purkinje cells of the cerebellar cortex. The exact involvement of cerebellar cortical input into the IN during eyeblink expression is not clear. While it is known that the application of gamma-aminobutyric acid-A (GABA(A)) agonists and antagonists affects the performance of classically conditioned eyeblinks, the effects of these drugs on IN neurons in vivo are not known. The purpose of the present study was to measure the effects of muscimol and picrotoxin on the expression of conditioned eyeblinks and the activity of IN cells simultaneously. Injections of muscimol abolished conditioned responses and either silenced or diminished the activity of IN cells. Two injections were administered in each picrotoxin experiment. The first injection of picrotoxin slightly modified the timing and amplitude of the eyeblink, produced mild tonic eyelid closure, increased tonic activity of IN cells, and reduced the amplitude of the neural responses. The second injection of picrotoxin abolished conditioned responses, further increased tonic eyelid closure, dramatically elevated the tonic activity of IN cells, and in most cases, abolished neuronal responses. These results demonstrate that both GABA(A)-mediated inactivation and tonic up-regulation of IN cells can interrupt the expression of conditioned eyeblinks and that this behavioral effect is accompanied by the suppression of the neuronal activity correlates of the conditioned stimulus and response.     

Rapid transfer of training occurs when direct mossy fiber stimulation is used as a conditioned stimulus for classical eyelid conditioning

Fifteen rabbits were classically conditioned using stimulation of the right pontine nuclei as a conditioned stimulus and a corneal airpuff as an unconditioned stimulus. After conditioning criterion was reached, the stimulation conditioned stimulus was transferred to the left pontine nuclei and the rate of conditioned response acquisition observed. Our results indicate that when electrodes were placed symmetrically in the right and left pontine nuclei, extremely rapid transfer of training occurred. Together with previous data, the present data supply further indirect evidence that the site of neural plasticity which underlies acquisition and retention of classically conditioned skeletal muscle responses is located efferent to the pontine nuclei, namely in the cerebellum.     

Classically conditioned withdrawal reflex in cerebellar patients. 2. Impaired unconditioned responses

The study addresses the issue of the role of the cerebellum in human withdrawal-reflex conditioning by comparing data from patients with pure cerebellar diseases (CBL, n = 10) and from cerebellar patients showing additional extracerebellar symptoms (CBL+, n = 10) with those from 11 control subjects (CTRL). During recording sessions, the standard delay-conditioning paradigm with paired-trials was used with tone as the conditioned stimulus (CS). Parameters of the conditioned muscle responses are analyzed in an accompanying paper. Here, we focus on the unconditioned muscle response. A train of current pulses (unconditioned stimulus, US) evoked a lower-limb withdrawal reflex (unconditioned response, UR), which was recorded electromyographically from leg muscles. During the recording sessions with CTRL subjects, UR amplitudes decayed from initially 100% to approximately 50% at the end of the session. This type of decay was clearly less pronounced in the CBL group and minimal in the CBL+ group. Furthermore, the CBL group exhibited UR onsets that were delayed by 20 ms compared with those from CTRL subjects. Although the ranges of measurements characterizing the URs of a given cerebellar patient tested in the paired-trial paradigm overlapped with those of control subjects, the statistically significant differences observed at the group level suggest deficits in the performance of the reflex responses. The delayed URs in patients and the different type of decay of UR amplitudes in repetitively evoked withdrawal reflexes constitute evidence that the cerebellum is critically involved in the control of these UR parameters.     

Dissociaton of conditioned eye and limb responses in the cerebellar interpositus

Thompson and colleagues have demonstrated that the lateral interpositus nucleus of the cerebellum is the essential locus for the classical conditioning of the somatic eyeblink response. Preliminary studies reported that lesioning the cerebellar interpositus nucleus ipsilateral to the side of training also appears to abolish conditioned limb flexion responses. Previous studies have suggested that the interpositus nucleus is somatotopically organized with the eye being represented laterally and the hindlimb medially. Presently, we employed a double dissociation paradigm to examine the effects of muscimol (a GABA(A) agonist) injections on eyeblink versus limb flexion conditioned responses in the ipsilateral cerebellar interpositus nucleus of New Zealand white rabbits. For eyeblink conditioning, the conditioned stimulus (CS) was a 14-V lamp bulb and the unconditioned stimulus (US) was a 3-psi corneal airpuff to the left eye. For limb flexion conditioning, the CS was a 1-kHz, 85-95 dB SPL tone and the US was a 3- to 5-mA shock to the upper left hindlimb. Upon training on both responses to a 60-100% criterion, the rabbits were then tested on eyeblink and limb flexion responses after injections of muscimol (0.1-0.3 mul of a 0.01- to 1.0-M solution) into either the lateral (eyeblink) or medial (limb flexion) interpositus nucleus. We have been able to successfully decrease or abolish the percent conditioned responses (CRs) of both the eyeblink and limb flexion conditioning selectively without affecting the other. These results thus lend further support for the notion of the existence of a somatotopic map in the interpositus nucleus for learning.     

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.     

Reflex facilitation of the nictitating membrane response remains after cerebellar lesions

Reflex facilitation and associated properties were investigated during classical conditioning of the nictitating membrane (NM) response in rabbit. In the first experiment, the role of the cerebellum was examined by comparing the unconditioned responses of animals with bilateral lesions of the deep cerebellar nuclei with those of operated controls during counterbalanced tone/light (T/L) discrimination training. Both T and L facilitated unconditioned NM responses when used as the CS+ (conditioned stimulus), but neither facilitated when used as the CS-. There were no significant differences in the amount of reflex facilitation exhibited by animals with lesions compared with control animals. Animals with lesions, however, failed to acquire conditioned responses after 10 days of training, whereas all control animals met acquisition criterion within 4 days. In the second experiment, reflex facilitation was shown to decrement in a stimulus-specific manner when nonreinforced presentations of an auditory stimulus were given. The discussion of results focuses on the relation between reflex facilitation and classical conditioning in terms of behavioral properties and underlying neural systems.     

Lock-and-key mechanisms of cerebellar memory recall based on rebound currents

A basic question for theories of learning and memory is whether neuronal plasticity suffices to guide proper memory recall. Alternatively, information processing that is additional to readout of stored memories might occur during recall. We formulate a "lock-and-key" hypothesis regarding cerebellum-dependent motor memory in which successful learning shapes neural activity to match a temporal filter that prevents expression of stored but inappropriate motor responses. Thus, neuronal plasticity by itself is necessary but not sufficient to modify motor behavior. We explored this idea through computational studies of two cerebellar behaviors and examined whether deep cerebellar and vestibular nuclei neurons can filter signals from Purkinje cells that would otherwise drive inappropriate motor responses. In eyeblink conditioning, reflex acquisition requires the conditioned stimulus (CS) to precede the unconditioned stimulus (US) by >100 ms. In our biophysical models of cerebellar nuclei neurons this requirement arises through the phenomenon of postinhibitory rebound depolarization and matches longstanding behavioral data on conditioned reflex timing and reliability. Although CS–US intervals <100 ms may induce Purkinje cell plasticity, cerebellar nuclei neurons drive conditioned responses only if the CS–US training interval was >100 ms. This bound reflects the minimum time for deinactivation of rebound currents such as T-type Ca²⁺. In vestibulo-ocular reflex adaptation, hyperpolarization-activated currents in vestibular nuclei neurons may underlie analogous dependence of adaptation magnitude on the timing of visual and vestibular stimuli. Thus, the proposed lock-and-key mechanisms link channel kinetics to recall performance and yield specific predictions of how perturbations to rebound depolarization affect motor expression.     

Forelimb reflexes modulated by tonic neck positions in cats

The modulation of monosynaptic forelimb reflexes by tonic neck positions was investigated in cats with the head fixed.Lateral flexion of the body in a horizontal plane markedly facilitates reflexes of the deep radial nerve (DR) in the ipsilateral forelimb, while the antagonistic ulnar nerve (ULN) reflexes are strongly inhibited. Opposite effects are seen after contralateral body movement.Dorsiflexion of the body clearly increases DR-reflexes and exerts a reciprocal although more pronounced inhibition on ULN reflexes. Opposite effects appear after ventriflexion.The reflex modulation starts with head-body displacements of approximately 5° and increases with increasing angles. Furthermore reflex modulation does not depend on the intact cerebrum and cerebellum. The comparison of forelimb and hindlimb reflexes shows a decrease of the neck influences along the spinal cord.Supported by Sonderforschungsbereich Hirnforschung und Sinnes-physiology (SFB 70) der Deutschen Forschungsgemeinschaft (DFG).Essential parts of this paper will be submitted by A. F. as a thesis to the Faculty of Medicine, Freiburg/Breisgau, Germany.     

Electrophysiological evidence for formation of new corticorubral synapses associated with classical conditioning in the cat

The present study was performed to clarify whether or not structural plasticity of synaptic connections underlies classical conditioning mediated by the red nucleus (RN) in the cat. Conditioned forelimb flexion is established by pairing electrical conditioned stimuli (CS), applied to corticorubral fibers at the cerebral peduncle (CP), with a forelimb skin shock (the unconditioned stimulus, US), but not by applying the CS alone or by pairing the CS and US at random intervals. In our previous study, it was shown that the firing probability of rubrospinal neurons (RN neurons) in response to the CS was well correlated with acquisition of the conditioned forelimb flexion and that the primary site of neural change underlying establishment of the conditioned forelimb flexion was suggested to be at corticorubral synapses. In the present study, we investigated corticorubral excitatory postsynaptic potentials evoked by CP stimulation (CP-EPSPs), in order to identify the neuronal mechanism underlying establishment of classical conditioning. In normal cats, CP-EPSPs had a typical slow-rising phase, which has been attributed to the distal location of corticorubral synapses on the dendrites of RN neurons. In contrast, in animals that received paired conditioning, subsequent CP stimulation evoked potentials with a fast-rising time course. In control groups of cats that received CS alone, CS randomly paired with the US, or only the same surgical operations as the conditioned animals, most of the CP-EPSPs displayed slow-rising EPSPs that similar to those observed in normal cats. The mean time from onset to peak of the potentials in the conditioned animals was significantly shorter than that seen in other groups. Therefore, the appearance of a fast-rising potential correlates well with acquisition of the conditioned forelimb flexion. The amplitude of the fast-rising potential was gradually changed with stimulus intensity. It had a short onset latency following CP stimulation (0.9 ms), which was similar to that of the slow-rising EPSP in normal cats. It followed high-frequency stimulation up to 100 Hz. These results suggest that the newly appearing, fast-rising potential was a monosynaptically evoked EPSP. Fast-rising EPSPs were also induced by stimulation of the sensorimotor cortex (SM). Since the SM-EPSP was occluded by the CP-EPSP, the SM cortex is, at least in part, a likely source of fast-rising EPSPs. Fast-rising SM-EPSPs were also observed at the unitary level. The SM-EPSPs in the conditioned animals exhibited somatotopical representation in their cortical origin, as has been described in normal cats. The electrotonic length was calculated from the voltage transient responses to current steps injected into the RN neurons. There was no concomitant change in the electrotonic length following the classical conditioning. Furthermore, the fastrising EPSPs were often observed as if they were superposed on the slow-rising EPSPs that were observed in normal animals. These observations suggest that the appearance of fast-rising EPSPs is due to the formation of new corticorubral synapses on the somata or the proximal dendrites of the RN neurons, and not as a result of a reduction in the electrotonic length of the RN neurons. The present study provides further evidence that this type of structural plasticity of synaptic connections underlies establishment of the classically conditioned forelimb flexion.     

Properties of conditioned abducens nerve responses in a highly reduced in vitro brain stem preparation from the turtle

Previous work suggested that the cerebellum and red nucleus are not necessary for the acquisition, extinction, and reacquistion of the in vitro classically conditioned abducens nerve response in the turtle. These findings are extended in the present study by obtaining conditioned responses (CRs) in preparations that received a partial ablation of the brain stem circuitry. In addition to removing all tissue rostral to and including the midbrain and cerebellum, a transection was made just caudal to the emergence of the IXth nerve. Such ablations result in a 4-mm-thick section of brain stem tissue that functionally eliminates the sustained component of the unconditioned response (UR) while leaving only a phasic component. We refer to this region of brain stem tissue caudal to the IXth nerve as the "caudal premotor blink region." Neural discharge was recorded from the abducens nerve following a single shock unconditioned stimulus (US) applied to the ipsilateral trigeminal nerve. When the US was paired with a conditioned stimulus (CS) applied to the posterior eighth, or auditory, nerve using a delay conditioning paradigm, a positive slope of CR acquisition was recorded in the abducens nerve, and CR extinction was recorded when the stimuli were alternated. Resumption of paired stimuli resulted in reacquisition. Quantitative analysis of the CRs in preparations in which the caudal premotor blink region had been removed and those with cerebellar/red nucleus lesions showed that both types of preparations had abnormally short latency CR onsets compared with preparations in which these regions were intact. Preparations with brain stem transections had significantly earlier CR offsets as more CRs terminated as short bursts when compared with intact or cerebellar lesioned preparations. These data suggest that a highly reduced in vitro brain stem preparation from the turtle can be classically conditioned. Furthermore, the caudal brain stem is not a site of acquisition in this reduced preparation, but it contributes to the sustained activity of both the UR and CR. Finally, the unusually short CR onset latencies following lesions to the cerebellum are not further exacerbated by removal of the caudal brain stem. These studies suggest that convergence of CS and US synaptic inputs onto the abducens nerve reflex circuitry may underlie acquisition in this reduced preparation, but that mechanisms that control learned CR timing arise from the cerebellorubral system.     

The role of the cerebellum in classical conditioning of discrete behavioral responses

The cerebellum and its associated circuitry constitutes the entire essential neuronal system for classical conditioning of eye-blink and other discrete responses (e.g. limb flexion) learned with an aversive unconditioned stimulus (US) using the standard delay paradigm where the conditioned stimulus (CS) and the US coterminate. Evidence reviewed here strongly supports the following conclusions. The CS pathway involves sensory relay nuclei projections to the pontine nuclei and its mossy fiber projections to the cerebellar cortex and nuclei. The US pathway involves activation of the inferior olive (dorsal accessory olive for eye blink) and its climbing fiber projections to the cerebellar cortex and nuclei. The conditioned response (CR) pathway involves the cerebellar interpositus nucleus, the superior cerebellar peduncle pathway to the magnocellular red nucleus and rubral projections to premotor and motor nuclei generating the behavioral response. Anatomical data, neuronal unit recordings, electrical stimulation, lesions and methods of reversible inactivation all strongly support the hypothesis that the essential memory trace for the learning of these discrete conditioned responses is formed and stored in the cerebellar interpositus nucleus. Neuronal/synaptic plasticity is also established in the cerebellar cortex in this form of learning but the role of the cortex is less clear. We argue that the cortex plays a key role in normal acquisition and adaptive timing of the conditioned response, under certain circumstances, but it remains unclear exactly what features of conditioning are being encoded in the cerebellar cortex in this basic form of associative learning and memory.     

Effect of experimentally induced heart disease on conditioned reflexes inhibiting respiration and the activity of the heart

Summary A reduction of the unconditioned reflex, which caused apnea and bradycardia was noted in animals with compenraten expenmental setic stenosis. The sound and light conditioned reflexes which were formed on its base were, likewise, reduced. The process of formation of the new conditioned reflexes on the base of the above unconditioned inflex became more difficult. These experimental data were evaluated in view of I. P. Pavlov's conception of the closure of the conditioned association between the cortical representation of unconditioned reflex and the cortical representation of the corresponding conditioned stimulant. It may be suggested that the reduction of the above unconditioned reflex in heart defect causes decreased excitability of its cortical representation. As a result of this the power of the process of excitation is decreased, the existing temporary associations are disturbed to a certain degree and the formation of new temporary associations becomes more difficult. The adaptive significance of reduction of unconditioned and conditioned reflexes, which inhibit the heart activity consists (in condition of heart defect) in the fact that it more or less guarantees the vitally important continuity of the compensatory hyperfunction of the heart.Presented by V. V. Parin, Member AMN SSSR     

Classically conditioned withdrawal reflex in cerebellar patients. 1. Impaired conditioned responses

The role of the cerebellum in the classically conditioned, human lower-limb-withdrawal reflex was studied in ten patients with pure cerebellar diseases (CBL), ten patients showing additional extracerebellar symptoms (CBL+), and in 11 sex- and age-matched normal controls (CTRL). Where conditioning was successful, the electrically evoked, unconditioned response was preceded by a tone-conditioned response (CR). CR incidence was variable, with best results in the CTRL, significantly less in CBL, and lowest in CBL+. Although CRs could be established in subjects in all groups, a continuous increase in the CR incidence in the course of the recording session was observed primarily in CTRL. In CBL and CBL+, such a characteristic reflex acquisition was rather the exception. CR onsets in CBL were within the range of those in CTRL, but CR amplitude was significantly lower in CBL. Cerebellar patients with circumscribed lesions behaved differently in our motor-learning paradigm, depending on the lesion site. Patients suffering from pathology of the posterior inferior cerebellum showed a mean CR incidence within the lower range of CTRL. In contrast, if the anterior and superior cerebellum was affected, few or even no CRs were observed. Our findings thus provide evidence that the human cerebellum is required for the acquisition and the retention of this specific conditioned limb-withdrawal reflex. In particular, anterior and superior parts of the cerebellum appear to be involved. Thus, an expansion of the current concept of clinically based, functional compartmentalization is suggested, such that anterior and superior cerebellar regions must be intact to establish plastic changes required for the acquisition of the conditioned withdrawal response.