Germund Hesslow

Acquisition, Extinction, and Reacquisition of a Cerebellar Cortical Memory Trace

Associative learning in the cerebellum underlies motor memories and probably also cognitive associations. Pavlovian eyeblink conditioning, a widely used experimental model of such learning, depends on the cerebellum, but the memory locus within the cerebellum as well as the underlying mechanisms have remained controversial. To date, crucial information on how cerebellar Purkinje cells change their activity during learning has been ambiguous and contradictory, and there is no information at all about how they behave during extinction and reacquisition. We have now tracked the activity of single Purkinje cells with microelectrodes for up to 16 h in decerebrate ferrets during learning, extinction, and relearning. We demonstrate that paired peripheral forelimb and periocular stimulation, as well as paired direct stimulation of cerebellar afferent pathways (mossy and climbing fibers) consistently causes a gradual acquisition of an inhibitory response in Purkinje cell simple spike firing. This conditioned cell response has several properties that matches known features of the behavioral conditioned response. The response latency varies with the interstimulus interval, and the response maximum is adaptively timed to precede the unconditioned stimulus. Across training trials, it matches behavioral extinction to unpaired stimulation and also the substantial savings that occur when paired stimulation is reinstated. These data suggest that many of the basic behavioral phenomena in eyeblink conditioning can be explained at the level of the single Purkinje cell.

Cerebellum and conditioned reflexes

The central assumption of existing models of motor learning in the cerebellum is that cerebellar mossy fibres signal information about the context in which a movement is to be performed and climbing fibres signal in relation to a movement error. This leads to changes in the responsiveness of Purkinje cells, which on the next occasion will generate a corrected output in a given context. Support for this view has come mainly from work on adaptation of the vestibulo-ocular reflex. The discovery that classically conditioned eyeblink responses depend critically on the cerebellum offers the possibility to study the learning of a novel behaviour, rather than modification of an existing reflex. After repeated pairing of a neutral stimulus, such as a tone, with a blink-eliciting stimulus, the tone will acquire the ability to elicit a blink on its own. We review evidence from studies employing a wide variety of techniques that the cerebellum is critical in this type of learning as well as evidence that mossy and climbing fibres have roles assigned to them in cerebellar learning models.

Correspondence between climbing fibre input and motor output in eyeblink-related areas in cat cerebellar cortex.

The purpose of the present work was to identify sites in the cerebellar cortex which are likely to control eyeblink. This work was motivated by findings suggesting that the cerebellum is involved in the learning and/or performance of the classically conditioned eyeblink response. The identification was based on climbing fibre input to the cortex and on the effects of electrical stimulation of the cerebellar cortex in cats decerebrated rostral to the red nucleus. The cerebellar surface was searched for areas receiving short latency climbing fibre input on periorbital electrical stimulation. Four such areas were found in the c1 and c3 zones of lobules VI and VII in the anterior lobe of the cerebellum and in the c3 zone in the paramedian lobule. Electrical stimulation of the cerebellar cortex with trains (150‐400 Hz) of at least 10 ms duration evoked two types of EMG response in the orbicularis oculi muscle. An early response, time‐locked to the onset of the stimulation, was unrelated to climbing fibre input and a delayed response, time‐locked to the termination of the stimulation, could only be evoked from areas which received short latency climbing fibre input from the eye, that is, the c1 and c3 zones. The delayed responses had long latencies (up to 50 ms) after the termination of the stimulus train and could be delayed further by prolonging the stimulation. Both types of response were abolished by injections of small amounts of lignocaine into the brachium conjunctivum. A number of characteristics of the delayed responses are described. They could be inhibited by a further shock to the same area of the cerebellar cortex. Their latency could be increased by increasing the stimulation frequency. The period between stimulation and appearance of the response often showed a decrease in spontaneous EMG activity. There was a close topographical correspondence between input and output. Delayed responses could be evoked from all four of the areas in the c1 and c3 zones which have climbing fibre input from the periorbital area. They could not be evoked from other areas. In contrast, early responses were only evoked from areas without such climbing fibre input. It is proposed that the delayed responses were generated by activation of Purkinje cell axons leading to hyperpolarization and a subsequent rebound depolarization and activation of cells in the interpositus nucleus. The cortical areas are therefore probably involved in the control of the orbicularis oculi muscle.(ABSTRACT TRUNCATED AT 400 WORDS)

Inhibition of classically conditioned eyeblink responses by stimulation of the cerebellar cortex in the decerebrate cat.

The purpose of the present study was to test the hypothesis that neurones in the anterior interpositus nucleus, under the control of Purkinje cells in the c1 and c3 zones of the cerebellar cortex, exert some control over classically conditioned responses. In particular, the experiments were designed to determine whether the cerebellar control of conditioned and unconditioned responses is different. The experiments were performed on cats decerebrated rostral to the red nucleus under halothane anaesthesia. The cats were conditioned using either a 1000 Hz tone or trains of stimuli through the skin of the proximal forelimb as the conditioned stimulus, and periorbital electrical stimulation as the unconditioned stimulus. A large proportion of the animals acquired conditioned responses at normal rates. It could be shown that these were true conditioned responses and did not result from sensitization or pseudoconditioning. For instance, unpaired presentations of conditioned and unconditioned stimuli caused rapid extinction. Cerebellar areas controlling eyeblink were identified by recording climbing fibre responses in the cerebellar cortex and recording EMG activity in the eyelid evoked by stimulation of the cerebellar cortex. When single shocks of 40‐70 microA were applied to these areas during the emission of conditioned eyeblink responses, the latter were strongly inhibited. The inhibition had a latency of about 10 ms and a duration of 25‐75 ms. It was shown that this inhibition of the conditioned responses was topographically specific and could only be evoked from cortical sites identified as controlling eyeblink. Stimulation of the periphery of an eyeblink area caused little or no inhibition. The effect of cortical stimulation on unconditioned reflex responses in the orbicularis oculi muscle was also tested. Some inhibition of unconditioned responses was observed, but quantitative analysis showed that this inhibition was considerably weaker than the corresponding inhibition of conditioned responses. The magnitude of the inhibition was determined for unconditioned responses of different sizes including responses which were weaker than the conditioned responses. It is concluded that conditioned eyeblink responses are under strong cerebellar control from areas in the c1 and c3 zones receiving climbing fibre input from the periorbital area. This effect is not likely to be due to a reduction in the background facilitation of facial motoneurones. In contrast, the weak inhibition of the unconditioned response was probably due to this mechanism. The results, therefore, suggest that the conditioned responses are dependent on the cerebellum in a way that is not true of unconditioned responses.

Inhibition of the inferior olive during conditioned responses in the decerebrate ferret

Output from the interpositus nucleus can inhibit the inferior olive, probably via the GABA-ergic nucleo-olivary pathway. It has been suggested that the function of this inhibition might be to regulate synaptic plasticity resulting from parallel fibre/climbing fibre interaction in cerebellar Purkinje cells, by providing negative feedback information to the olive. Thus, when a learned response, generated by the interpositus nucleus, reaches a sufficient amplitude, the olive would be inhibited and further learning blocked. This suggestion was tested in a classical conditioning paradigm. Decerebrate ferrets were trained using electrical skin stimulation of the forelimb as the conditioned stimulus (CS) and periorbital stimulation as the unconditioned stimulus (US). Climbing fibre responses evoked in Purkinje cells by the US were recorded as surface field potentials in the part of the c3 zone controlling eyeblink. It was found that the CS did not inhibit the olive at the beginning of training, but when conditioned responses were large, the olive was inhibited by the CS in some animals. After a number of unpaired CS presentations, which caused extinction of the conditioned response, the inhibition disappeared. The size of individual conditioned responses correlated negatively with the size of the climbing fibre responses evoked by the US. Climbing fibre responses evoked by direct stimulation of the olive were also inhibited. It was concluded that cerebellar output during performance of a conditioned response inhibits the inferior olive. The results thus support the hypothesis of a cerebellar locus of conditioning and are consistent with the proposed role of cerebello-olivary inhibition.

Suppression of cerebellar Purkinje cells during conditioned responses in ferrets.

Decerebrate ferrets were conditioned, using electrical stimulation of the forelimb as conditioned stimulus and periorbital stimulation as unconditioned stimulus, until they produced conditioned eyeblink responses. The latency of these was 125-250 ms. Microelectrode recordings were made from single Purkinje cells in an eyeblink controlling area in the c3 zone of the cerebellar cortex. Whereas Purkinje cells in animals, which had only received unpaired stimulus presentations, responded weakly or not at all to the conditioned stimulus, some cells in conditioned animals responded with a powerful suppression of simple spike firing. The latency of this suppression was 50-200 ms. The results support the hypothesis that classical conditioning involves plastic changes in cerebellar Purkinje cells.

Learned movements elicited by direct stimulation of cerebellar mossy fiber afferents.

Definitive evidence is presented that the conditioned stimulus (CS) in classical conditioning reaches the cerebellum via the mossy fiber system. Decerebrate ferrets received paired forelimb and periocular stimulation until they responded with blinks to the forelimb stimulus. When direct mossy fiber stimulation was then given, the animals responded with conditioned blinks immediately, that is, without ever having been trained to the mossy fiber stimulation. Antidromic activation was prevented by blocking mossy fibers with lignocaine ventral to the stimulation site. It could be excluded that cerebellar output functioned as the CS. Analysis of latencies suggests that conditioned responses (CRs) are not generated by mossy fiber collaterals to the deep nuclei. Hence, the memory trace is probably located in the cerebellar cortex.

Cerebellar control of the inferior olive.

A subpopulation of neurones in the cerebellar nuclei projects to the inferior olive, the source of the climbing fibre input to the cerebellum. This nucleo-olivary projection follows the zonal and, probably also, the microzonal arrangement of the cerebellum so that closed loops are formed between the neurones in the olive, the cerebellar cortex and the nuclei. The nucleo-olivary pathway is GABAergic, but several investigators argue that its main effect is to regulate electrotonic coupling between cells in the inferior olive rather than inhibit the olive. However, there is now strong evidence that the nucleo-olivary fibres do inhibit the olive. Three functions have been suggested for this inhibition: (i) feedback control of background activity in Purkinje cells, (ii) feedback control of learning, and (iii) gating of olivary input in general. Evidence is consistent with (i) and (ii). Activity in the nucleo-olivary pathway suppresses both synaptic transmission and background activity in the olive. When learned blink responses develop, the blink related part of the olive is inhibited while blinks are produced. When the nucleo-olivary pathway is interrupted, there is a corresponding increase in complex spike discharge in Purkinje cells followed by a strong suppression of simple spike firing. Stimulation of the pathway has the opposite results. It is concluded that the nucleo-olivary fibres are inhibitory and that they form a number of independent feedback loops, each one specific for a microcomplex, that regulate cerebellar learning as well as spontaneous activity in the olivo-cerebellar circuit.

Evidence for a GABA-mediated cerebellar inhibition of the inferior olive in the cat

Summary1. Climbing fibres were activated by peripheral nerve stimulation at ‘high’ frequencies (>3 Hz) for 15–25 s and then at 0.9 Hz for about 1 min. The high frequency activation induced a post-conditioning inhibition, lasting up to about 1 min, of climbing fibre responses recorded from the cerebellar surface. 2. Electrolytic lesions were made in the superior cerebellar peduncle (brachium conjunctivum). After the lesion, the post-conditioning inhibition was completely eliminated. 3. Injections of the GABA-receptor blocker bicuculline methiodide into the inferior olive reversibly blocked the post-conditioning inhibition. 4. The results support the hypothesis proposed by Andersson and Hesslow (1987a), that post-conditioning inhibition is mediated by a GABA-ergic interposito-olivary pathway.

Do we need a concept of disease

The terms “health”, “disease” and “illness” are frequently used in clinical medicine. This has misled philosophers into believing that these concepts are important for clinical thinking and decision making. For instance, it is held that decisions about whether or not to treat someone or whether to relieve someone of moral responsibility depend on whether the person has a disease. In this paper it is argued that the crucial role of the ‘disease’ concept is illusory. The health/disease distinction is irrelevant for most decisions and represents a conceptual straightjacket. Sophisticated and mature clinical decision making requires that we free ourselves from the concept of disease.