Nadia L. Cerminara

Evidence that Climbing Fibers Control an Intrinsic Spike Generator in Cerebellar Purkinje Cells

It is well established that the climbing fiber (CF) input to a cerebellar Purkinje cell (PC) can exert a controlling influence on the background simple spike (SS) activity of the cell, in that repetitive stimulation of CFs causes a decrease in SS activity, and removal or inactivation of CFs is followed by a rise in activity. In the present study, the effects of inactivation of CFs in the short term and longer term (hours) were investigated in anesthetized rats to determine how the CFs control the PC SS activity. Inactivation of the CF input to a PC was accomplished by either reversibly inactivating with lignocaine or by microlesioning the inferior olive. Consistent with previous findings, CF removal caused a transformation of the PC firing pattern, with SSs discharging more regularly and rising to an exceptionally high level. In cases in which CF activity resumed, SS rate declined to control levels within a few seconds. However, with sustained CF inactivation (30 min to 5 hr), SS activity continues to rise progressively and develops an oscillating firing pattern, consisting of alternating bursts of high-frequency discharge at up to 100-150 Hz followed by 10-20 sec periods of electrical quiescence. No accompanying changes in the threshold for evoking SSs via the parallel fibers were seen to accompany the increases in tonic SS activity. We conclude that the increase in SS activity that follows CF inactivation could be caused by the removal of an inhibitory action that CFs exert on the intrinsic pacemaker present in PCs under normal conditions.

Redefining the cerebellar cortex as an assembly of non-uniform Purkinje cell microcircuits

The adult mammalian cerebellar cortex is generally assumed to have a uniform cytoarchitecture. Differences in cerebellar function are thought to arise primarily through distinct patterns of input and output connectivity rather than as a result of variations in cortical microcircuitry. However, evidence from anatomical, physiological and genetic studies is increasingly challenging this orthodoxy, and there are now various lines of evidence indicating that the cerebellar cortex is not uniform. Here, we develop the hypothesis that regional differences in properties of cerebellar cortical microcircuits lead to important differences in information processing.

An internal model of a moving visual target in the lateral cerebellum

In order to overcome the relatively long delay in processing visual feedback information when pursuing a moving visual target, it is necessary to predict the future trajectory of the target if it is to be tracked with accuracy. Predictive behaviour can be achieved through internal models, and the cerebellum has been implicated as a site for their operation. Purkinje cells in the lateral cerebellum (D zones) respond to visual inputs during visually guided tracking and it has been proposed that their neural activity reflects the operation of an internal model of target motion. Here we provide direct evidence for the existence of such a model in the cerebellum by demonstrating an internal model of a moving external target. Single unit recordings of Purkinje cells in lateral cerebellum (D2 zone) were made in cats trained to perform a predictable visually guided reaching task. For all Purkinje cells that showed tonic simple spike activity during target movement, this tonic activity was maintained during the transient disappearance of the target. Since simple spike activity could not be correlated to eye or limb movements, and the target was familiar and moved in a predictable fashion, we conclude that the Purkinje cell activity reflects the operation of an internal model based on memory of its previous motion. Such a model of the targets motion, reflected in the maintained modulation during the targets absence, could be used in a predictive capacity in the interception of a moving object.

Mechanisms of synchronous activity in cerebellar Purkinje cells

Complex spike synchrony is thought to be a key feature of how inferior olive climbing fibre afferents make their vital contribution to cerebellar function. However, little is known about whether the other major cerebellar input, the mossy fibres (which generate simple spikes within Purkinje cells, PCs), exhibit a similar synchrony in impulse timing. We have used a multi‐microelectrode system to record simultaneously from two or more PCs in the posterior lobe of the ketamine/xylazine‐anaesthetized rat to examine the relationship between complex spike and simple spike synchrony in PC pairs located mainly in the A2 and C1 zones in crus II and the paramedian lobule. PC pairs displaying correlations in the occurrence of their complex spikes (coupled PCs) were usually located in the same zone and were also more likely to exhibit correlations in the timing of their spontaneous simple spikes and associated pauses in activity. In coupled PCs, synchrony in both complex spike and simple spike activity was enhanced and the relative timing in the occurrence of complex spikes could be altered by peripheral stimulation. We conclude that the functional coupling between PC pairs in their complex spike and simple spike activity can be significantly modified by sensory inputs, and that mechanisms besides electrotonic coupling are involved in generating PC synchrony. Synchronous activity in multiple PCs converging onto the same cerebellar nuclear cells is likely to have a significant impact on cerebellar output that could form important timing signals to orchestrate coordinated movements.

Behavioural Significance of Cerebellar Modules

A key organisational feature of the cerebellum is its division into a series of cerebellar modules. Each module is defined by its climbing input originating from a well-defined region of the inferior olive, which targets one or more longitudinal zones of Purkinje cells within the cerebellar cortex. In turn, Purkinje cells within each zone project to specific regions of the cerebellar and vestibular nuclei. While much is known about the neuronal wiring of individual cerebellar modules, their behavioural significance remains poorly understood. Here, we briefly review some recent data on the functional role of three different cerebellar modules: the vermal A module, the paravermal C2 module and the lateral D2 module. The available evidence suggests that these modules have some differences in function: the A module is concerned with balance and the postural base for voluntary movements, the C2 module is concerned more with limb control and the D2 module is involved in predicting target motion in visually guided movements. However, these are not likely to be the only functions of these modules and the A and C2 modules are also both concerned with eye and head movements, suggesting that individual cerebellar modules do not necessarily have distinct functions in motor control.

Systematic regional variations in Purkinje cell spiking patterns

In contrast to the uniform anatomy of the cerebellar cortex, molecular and physiological studies indicate that significant differences exist between cortical regions, suggesting that the spiking activity of Purkinje cells (PCs) in different regions could also show distinct characteristics. To investigate this possibility we obtained extracellular recordings from PCs in different zebrin bands in crus IIa and vermis lobules VIII and IX in anesthetized rats in order to compare PC firing characteristics between zebrin positive (Z+) and negative (Z−) bands. In addition, we analyzed recordings from PCs in the A2 and C1 zones of several lobules in the posterior lobe, which largely contain Z+ and Z− PCs, respectively. In both datasets significant differences in simple spike (SS) activity were observed between cortical regions. Specifically, Z− and C1 PCs had higher SS firing rates than Z+ and A2 PCs, respectively. The irregularity of SS firing (as assessed by measures of interspike interval distribution) was greater in Z+ bands in both absolute and relative terms. The results regarding systematic variations in complex spike (CS) activity were less consistent, suggesting that while real differences can exist, they may be sensitive to other factors than the cortical location of the PC. However, differences in the interactions between SSs and CSs, including the post-CS pause in SSs and post-pause modulation of SSs, were also consistently observed between bands. Similar, though less strong trends were observed in the zonal recordings. These systematic variations in spontaneous firing characteristics of PCs between zebrin bands in vivo, raises the possibility that fundamental differences in information encoding exist between cerebellar cortical regions.

Structural Basis of Cerebellar Microcircuits in the Rat

The topography of the cerebellar cortex is described by at least three different maps, with the basic units of each map termed “microzones,” “patches,” and “bands.” These are defined, respectively, by different patterns of climbing fiber input, mossy fiber input, and Purkinje cell (PC) phenotype. Based on embryological development, the “one-map” hypothesis proposes that the basic units of each map align in the adult animal and the aim of the present study was to test this possibility. In barbiturate anesthetized adult rats, nanoinjections of bidirectional tracer (Retrobeads and biotinylated dextran amine) were made into somatotopically identified regions within the hindlimb C1 zone in copula pyramidis. Injection sites were mapped relative to PC bands defined by the molecular marker zebrin II and were correlated with the pattern of retrograde cell labeling within the inferior olive and in the basilar pontine nuclei to determine connectivity of microzones and patches, respectively, and also with the distributions of biotinylated dextran amine-labeled PC terminals in the cerebellar nuclei. Zebrin bands were found to be related to both climbing fiber and mossy fiber inputs and also to cortical representation of different parts of the ipsilateral hindpaw, indicating a precise spatial organization within cerebellar microcircuitry. This precise connectivity extends to PC terminal fields in the cerebellar nuclei and olivonuclear projections. These findings strongly support the one-map hypothesis and suggest that, at the microcircuit level of resolution, the cerebellar cortex has a common plan of spatial organization for major inputs, outputs, and PC phenotype.

Consensus paper: current views on the role of cerebellar interpositus nucleus in movement control and emotion.

In the present paper, we examine the role of the cerebellar interpositus nucleus (IN) in motor and non-motor domains. Recent findings are considered, and we share the following conclusions: IN as part of the olivo-cortico-nuclear microcircuit is involved in providing powerful timing signals important in coordinating limb movements; IN could participate in the timing and performance of ongoing conditioned responses rather than the generation and/or initiation of such responses; IN is involved in the control of reflexive and voluntary movements in a task- and effector system-dependent fashion, including hand movements and associated upper limb adjustments, for quick effective actions; IN develops internal models for dynamic interactions of the motor system with the external environment for anticipatory control of movement; and IN plays a significant role in the modulation of autonomic and emotional functions.

The lateral cerebellum and visuomotor control.

The lateral cerebellum receives an abundance of visual input providing the link between visual and motor control centers. In this review we discuss experiments designed to increase our understanding of how visual inputs to the cerebellum are arranged in relation to the zonal organization of the cerebellar cortex, and how visual inputs are utilized to assist in the regulation of a visually guided movement. On the basis of anatomical and physiological characteristics our findings indicate that the medial-most folium in crus I of the cat lateral cerebellum can be subdivided into at least three functionally distinct zones; from lateral to medial along the length of the folium these correspond to zones D(1), lateral C(3) and C(2). Each zone displays clear differences in olivo-cortico-nuclear connectivity and in the anesthetized animal zones D(1) and C(2) both receive powerful visual inputs relayed via the climbing fiber system. Complementary experiments in awake behaving cats found that Purkinje cells located in the D(1) and D(2) zones of crus I exhibit changes in simple spike discharge time locked to target motion during a visually guided reaching task. These changes were unaffected by temporary visual denial of the target, raising the possibility that internally generated feedforward visuomotor control mechanisms are operating, in which a predictive model of the targets motion has been constructed by the CNS.

Purkinje cells in the lateral cerebellum of the cat encode visual events and target motion during visually guided reaching.

In this study the receipt of visual information by the lateral cerebellum and its contribution to a motor output was studied using single unit recording of cerebellar cortical neurones in cats trained to perform visually guided reaching. The activity of Purkinje cells and other cortical neurones in the lateral cerebellum was investigated in relation to various aspects of the task, such as visual events, parameters of target movement, and limb and eye movements. Two‐thirds (66%) of Purkinje cells tested could signal simple visual events, such as a flash of light. Neurones were also capable of detecting other less potent, but behaviourally important visual events, such as a ‘GO’ signal (LED brightening). Half of the cells tested were responsive to the on‐going motion of the visual target, displaying tonically altered discharge rates for as long as it was moving, and a ‘preferred’ target velocity. A small proportion of cells showed short latency visual modulation that persisted during the forelimb reach. Anatomical tracing studies confirmed that the recordings were obtained from the D1 zone of crus I. In summary, cells in this region of lateral cerebellar cortex perform simple visual functions, such as event detection, but also more complex visual functions, such as encoding parameters of target motion, and their visual responsiveness is appropriate for a role in accurate visually guided reaching to a moving target.