Cerebellar Output Research Articles

Cerebellar Output in Zebrafish: An Analysis of Spatial Patterns and Topography in Eurydendroid Cell Projections

The cerebellum is a brain region responsible for motor coordination and for refining motor programs. While a great deal is known about the structure and connectivity of the mammalian cerebellum, fundamental questions regarding its function in behavior remain unanswered. Recently, the zebrafish has emerged as a useful model organism for cerebellar studies, owing in part to the similarity in cerebellar circuits between zebrafish and mammals. While the cell types composing their cerebellar cortical circuits are generally conserved with mammals, zebrafish lack deep cerebellar nuclei, and instead a majority of cerebellar output comes from a single type of neuron: the eurydendroid cell. To describe spatial patterns of cerebellar output in zebrafish, we have used genetic techniques to label and trace eurydendroid cells individually and en masse. We have found that cerebellar output targets the thalamus and optic tectum, and have confirmed the presence of pre-synaptic terminals from eurydendroid cells in these structures using a synaptically targeted GFP. By observing individual eurydendroid cells, we have shown that different medial-lateral regions of the cerebellum have eurydendroid cells projecting to different targets. Finally, we found topographic organization in the connectivity between the cerebellum and the optic tectum, where more medial eurydendroid cells project to the rostral tectum while lateral cells project to the caudal tectum. These findings indicate that there is spatial logic underpinning cerebellar output in zebrafish with likely implications for cerebellar function.

Cerebellar cortical neuron responses evoked from the spinal border cell tract

Spinocerebellar systems are likely to be crucial for cerebellar hallmark functions such as coordination. However, in terms of cerebellar functional analyses, these are perhaps among the least explored systems. The aim of the present study is to achieve activation of a single component of the spinocerebellar systems and to explore to what extent it can influence the spike output of granule cells, Golgi cells, molecular layer (ML) interneurons (stellate and basket cells) and Purkinje cells (PCs). For this purpose, we took advantage of a unique arrangement discovered in neuroanatomical studies, in which the spinal border cell (SBC) component of the ventral spinocerebellar system was found to be the only spinocerebellar tract which ascends in the contralateral lateral funiculus (coLF) and have terminations in sublobulus C1 of the paramedian lobule in the posterior cerebellum. Using electrical stimulation of this tract, we find a subset of the cerebellar cortical neurons in this region to be moderately or powerfully activated. For example, some of our granule cells displayed high intensity responses whereas the majority of the granule cells displayed no response at all. The finding that more than half of the PCs were activated by stimulation of the SBC tract indicated that this system is capable of directly influencing cerebellar cortical output. The implications of these findings for the view of the integrative functions of the cerebellar cortex are discussed.

Medial cerebellar nuclear projections and activity patterns link cerebellar output to orofacial and respiratory behavior

There is ample evidence that the cerebellum plays an important role in coordinating both respiratory and orofacial movements. However, the pathway by which the cerebellum engages brainstem substrates underlying these movements is not well understood. We used tract-tracing techniques in mice to show that neurons in the medial deep cerebellar nucleus (mDCN) project directly to these putative substrates. Injection of an anterograde tracer into the mDCN produced terminal labeling in the ventromedial medullary reticular formation, which was stronger on the contralateral side. Correspondingly, injection of retrograde tracers into these same areas resulted in robust neuronal cell labeling in the contralateral mDCN. Moreover, injection of two retrograde tracers at different rostral–caudal brainstem levels resulted in a subset of double-labeled cells, indicating that single mDCN neurons collateralize to multiple substrates. Using an awake and behaving recording preparation, we show that spiking activity in mDCN neurons is correlated with respiratory and orofacial behaviors, including whisking and fluid licking. Almost half of the recorded neurons showed activity correlated with more than one behavior, suggesting that these neurons may in fact modulate multiple brainstem substrates. Collectively, these results describe a potential pathway through which the cerebellum could modulate and coordinate respiratory and orofacial behaviors.

Progressive limb ataxia following inferior olive lesions

Cerebellar climbing fibres originate in the inferior olive (IO). Temporary IO inactivation produces movement deficits. Does permanent inactivation produce similar deficits and, if so, do they recover? The excitotoxin, kainic acid, was injected into the rostral IO of three cats. Behaviour was measured during reaching and locomotion. Two cats were injected during the reaching task. Within minutes, grasping became difficult and the trajectories of the reaches showed higher arcing than normally seen. During locomotion, both cats showed head and trunk deviation to the injected side, walking paths curved to the injected side, and the paws were lifted higher than normal. Limbs contralateral to the injections became rigid. Within 1 day, posture had normalized, locomotion was unsteady and high lifting of the paws had reversed to a tendency to drag the dorsum of the paws. Passive body movement produced vestibular signs. Over a few days, locomotion normalized and vestibular signs disappeared. Reach trajectories were normal but grasping deficits persisted. Over the first week, the amplitude of limb lift during reaching and locomotion began to increase. The increase continued over time and, after several months, limb movements became severely ataxic. The effects followed the somatotopy of the rostral IO: a loss of cells in medial rostral IO only affected the forelimb, whereas a loss of cells in medial and lateral IO affected both forelimb and hindlimb. Deficits produced by IO lesions involve multiple mechanisms; some recover rapidly, some appear stable, and some worsen over time. The nature of the progressive deficit suggests a gradual loss of Purkinje cell inhibition on cerebellar nuclear cells.

Anatomical evidence for the involvement of medial cerebellar output from the interpositus nuclei in cognitive functions

Although the cerebellar interpositus nuclei are known to be involved in cognitive functions, such as associative motor learning, no anatomical evidence has been available for this issue. Here we used retrograde transneuronal transport of rabies virus to identify neurons in the cerebellar nuclei that project via the thalamus to area 46 of the prefrontal cortex of macaques in comparison with the projections to the primary motor cortex (M1). After rabies injections into area 46, many neurons in the restricted region of the posterior interpositus nucleus (PIN) were labeled disynaptically via the thalamus, whereas no neuron labeling was found in the anterior interpositus nucleus (AIN). The distribution of the labeled neurons was dorsoventrally different from that of PIN neurons labeled from the M1. This defines an anatomical substrate for the contribution of medial cerebellar output to cognitive functions. Like the dentate nucleus, the PIN has dual motor and cognitive channels, whereas the AIN has a motor channel only.

Lesion of the cerebellar paravermis increases dopamine D1 receptor levels in the contralateral striatum

Anatomical and biochemical findings have long suggested that a projection from the cerebellum to the basal ganglia exists, and recent findings proposed that the cerebellum influences glutamatergic striatal activity. We have previously shown that a complete, genetic, lack of Purkinje cells induces an upregulation of dopamine D1 receptors (DRD1) in the output of the basal ganglia, the substantia nigra pars reticulata. In this study, we produced a focal unilateral lesion in the cerebellar paravermal cortex and we studied the levels and distribution of dopamine receptors and transporters, with the use of in vitro receptor autoradiography. The lesion produced a statistically significant increase in DRD1 specific binding in the contralateral medial striatum and a bilateral decrease in dopamine transporter (DAT) levels in the dorsolateral striatum. Our finding of a DRD1 increase after disruption of the cerebellar corticonuclear projection suggests that the cerebellar output modulates the basal ganglia DRD1-mediated pathway.

Motor dysfunction and cerebellar Purkinje cell firing impairment in Ebf2 null mice

Mice deficient for the transcription factor Ebf2 lose a subset of Purkinje cells during development and have a hypotrophic cerebellar cortex. Related motor symptoms and the function of Purkinje cells surviving in the adult have not been studied so far. Ebf2 null mice presented locomotor impairment and a deficiency of motor coordination and motor learning. Ebf2 null Purkinje cells of the anterior lobe, relative to wild-type controls, were patch-clamp recorded in acutely prepared slices. While immature Purkinje cells (10-20 postnatal days) of Ebf2 null mice showed no significant difference relative to wild-types, in the adult they featured a higher input resistance, increased anomalous rectification, decreased first spike latency, higher initial firing frequency, lower voltage threshold and reduced afterhyperpolarizations and post-burst hyperpolarizations. These parameters indicate a difference in the response to both hyperpolarizing and depolarizing stimuli, corresponding to an altered cerebellar cortical output signaling. In contrast, adult climbing fibers attained a normal monoinnervation pattern and the parallel fiber-Purkinje cell synapse showed evoked postsynaptic currents and paired-pulse facilitation functionally indistinguishable from wild-type PCs. These results suggest that the motor deficits exhibited by Ebf2 null mice could be due, at least in part, to an impairment of the firing properties of surviving Purkinje cells. These findings indicate that Ebf2 is important for the development and maintenance of normal Purkinje cell discharge properties.

Cerebellar rTMS stimulation may induce prolonged clinical benefits in essential tremor, and subjacent changes in functional connectivity: An open label trial

BackgroundCerebello-thalamo-cortical (CTC) pathways dysfunction is involved in pathological oscillations causing tremor in essential tremor (ET). Low-frequency (1 Hz) repetitive transcranial magnetic stimulation (rTMS) of the cerebellum can effectively modulate the cerebellar output. ObjectiveAs one session of rTMS can induce a brief improvement, we hypothesized that repeated sessions might have a cumulative and potentially long-term therapeutic effect on ET. We assessed, in an open label trial, the efficacy of one-week rTMS treatment on tremor and on the motor-CTC dysfunction in ET patients. MethodsResting-state fMRI functional connectivity was used as an indicator of CTC network integrity in 11 ET patients and 11 healthy subjects. Resting-state fMRI connectivity was quantified at baseline in patients and control subjects between the cerebellum and the motor network, and between the cerebellum and the default brain network (DBN) taken as control. The fMRI study was repeated in patients after 5 days of bilateral 1 Hz rTMS applied to the posterior cerebellar cortex. Tremor was assessed clinically (Fahn–Tolosa–Marin scale) and quantified using electromyographic and accelerometric recordings at baseline (day 1, before the cerebellar stimulation) and after the end of the cerebellar stimulation period at day 5, day 12 and day 29. ResultsRepeated rTMS over the cerebellum significantly improved total and specific (tremor, drawing, functional disability) scores, and reduced tremor amplitude (P < 0.006). It also re-established the defective information processing in the CTC network (P(Δ|y) > 0.909), but not in the DBN. The effects persisted for 3 weeks after the last session. ConclusionCerebellar stimulation could be an effective treatment option for patients with severe essential tremor.

Diversity of Neuronal Elements and Circuitry in the Cerebellar Nuclei

The afferent and efferent synaptic connections of the cerebellar nuclei (CN) place them in a key position where they can integrate sensory signals with the output from cerebellar cortex and to provide the main efferent pathway of the cerebellum. While this conclusion can be derived based on purely anatomical knowledge, it remains unknown in which manner the CN contributes to the generation of cerebellar output signals that are involved in creating timing signals and temporal patterns. As a first step towards understanding the role neuronal circuits of the CN, the major CN neuronal types are now identified based on expression patterns of neurotransmitters (GABA and glycine) and characterized both in electrophysiological and morphological manner. The classification-likely to be refined in the future-consists of six types: four classes of projection and two classes of local neurons. The classification is a combination of electrophysiological and morphological methods with the expression pattern of GAD67 and GlyT2, markers for GABAergic and glycinergic neurons, respectively (Uusisaari et al. J Neurophysiol 97:901-911, 2007; Uusisaari and Knöpfel, Cerebellum 10(4):637-46, 2010).

The Cerebellum and Cerebello-Thalamo-Cortical Channels Contribute to New Learning and Long-Term Memory of Motor Skill

Lu and his research team used the retrograde transneuronal transport of rabies virus to identify neurons in the cerebellar nuclei that project via the thalamus to the cerebral cortex of macaques. In particular, projections from deep cerebellar nuclei to area 46 of the prefrontal cortex were compared with projections to the primary motor cortex (M1). They found that, after viral injections into area 46, many labeled neurons were observed in the ventral aspect of the Posterior Interpositus Nucleus (PIN), whereas no neuron labeling was found in the Anterior Interpositus Nucleus (AIN). In contrast, a number of labeled neurons were found in the dorsal portion of the PIN after viral injections into the M1. Additionally, neurons labeled from the M1 injections were also observed in the AIN [1-3]. These findings demonstrate that the cerebellar interpositus nuclei possess distinct outputs to the prefrontal cortex versus the M1, which potentially contribute to different aspects of behavioral functions such as different stages of the motor learning. Medial cerebellar outputs from the interpositus nuclei have been thought to be crucial for the associative motor learning. McCormick and Thompson have reported that lesions of the interpositus nuclei abolished the memorized eyeblink response; recordings from these nuclei have revealed neuronal activity to response learning [4]. Yet, clinical studies have reported that initial stage of trace eyeblink conditioning learning was impaired in patients with lesions in the PIN [5]. Imaging studies have also shown that the cerebellum play a key role in eyeblink conditioning [6-8]. Moreover, significant changes of positron emission tomography and regional cerebral blood flow in several areas, including the cerebellum and the prefrontal cortex, were observed during performance of associative motor learning task [9]. The above, taken together with the new medial cerebellar channels from the posterior interpositus nucleus to area 46 of the prefrontal cortex, it is reasonable to hypothesize that the cerebellum and cerebello-thalamocortical channels contribute to motor learning. The medial cerebellar output from the ventral PIN and the dorsal PIN/AIN play differential roles in various steps of the motor learning [5,10,11]. The dorsal aspect of the PIN has been thought to contribute to performance of memorized motor responses such as eyeblink and saccadic eye movements rather than to their initial learning [10, 11]. If this is the case, then impairment in the acquisition of the new learning by lesions of the PIN could be due to functional blockade of the ventral rather than dorsal portion of the PIN [5]. Furthermore, previous studies have shown evidence that the expression of representative motor memory formation genes was increased selectively in the AIN, but not in the PIN [12]. This finding strongly suggests that the AIN is involved in the storage and/or retrieval of long-term memory of motor learning. Moreover, many studies have indicated that the AIN is the site for long-term memory rather than new learning during practice of the eyeblink conditioning response. Examples include lesion [13-17]; inactivation [18,19]; electron microscopy [20]; functional magnetic resonance imaging [21]. Given the above, it is natural to raise a theory that the medial cerebellar output from the ventral PIN and the dorsal PIN/AIN plays a critical role in the new learning and long-term memory, respectively, during the motor learning processing. Interestingly, different cortical areas have been thought to be involved in these various steps of motor learning. A considerable theory for motor learning indicates that new learning is an explicit process originating in the prefrontal cortex, and that the long term memory for the learned motor skill becomes automatic and an implicit process formed in the motor cortex [22-24]. Thus, the disynaptic pathway from the ventral aspect of the PIN to area 46 may be involved in the initiation of the association of motor learning. After repeated practice, cerebellar channels to the M1 originating from the dorsal PIN/AIN turn to play a key role in the storage and/or retrieval of long-term memory, which makes the association automatic. Last, neuropsychological testing of patients with cerebellar lesions has revealed specific deficits in different faculties, including visual perception [25-31], short-term memory [32,33], or verbal fluency [34]. Moreover, clinical studies have shown that, in the brains of subjects with autism, the most consistent abnormalities are found in cerebellar Purkinje cells [35]. Accordingly, the questions of whether and how these cerebello-thalamo-cortical channels and their involvements in

Unravelling cerebellar pathways with high temporal precision targeting motor and extensive sensory and parietal networks

Increasing evidence has implicated the cerebellum in providing forward models of motor plants predicting the sensory consequences of actions. Assuming that cerebellar input to the cerebral cortex contributes to the cerebro-cortical processing by adding forward model signals, we would expect to find projections emphasising motor and sensory cortical areas. However, this expectation is only partially met by studies of cerebello-cerebral connections. Here we show that by electrically stimulating the cerebellar output and imaging responses with functional magnetic resonance imaging, evoked blood oxygen level-dependant activity is observed not only in the classical cerebellar projection target, the primary motor cortex, but also in a number of additional areas in insular, parietal and occipital cortex, including sensory cortical representations. Further probing of the responses reveals a projection system that has been optimized to mediate fast and temporarily precise information. In conclusion, both the topography of the stimulation effects and its emphasis on temporal precision are in full accordance with the concept of cerebellar forward model information modulating cerebro-cortical processing.

Historical Review of the Cerebellar Output: Motor and Cognitive Channels

Historical Review of the Cerebellar Output: Motor and Cognitive Channels

Defective dentate nucleus GABA receptors in essential tremor

The development of new treatments for essential tremor, the most frequent movement disorder, is limited by a poor understanding of its pathophysiology and the relative paucity of clinicopathological studies. Here, we report a post-mortem decrease in GABA(A) (35% reduction) and GABA(B) (22-31% reduction) receptors in the dentate nucleus of the cerebellum from individuals with essential tremor, compared with controls or individuals with Parkinson's disease, as assessed by receptor-binding autoradiography. Concentrations of GABA(B) receptors in the dentate nucleus were inversely correlated with the duration of essential tremor symptoms (r(2) = 0.44, P < 0.05), suggesting that the loss of GABA(B) receptors follows the progression of the disease. In situ hybridization experiments also revealed a diminution of GABA(B(1a+b)) receptor messenger RNA in essential tremor (↓27%). In contrast, no significant changes of GABA(A) and GABA(B) receptors (protein and messenger RNA), GluN2B receptors, cytochrome oxidase-1 or GABA concentrations were detected in molecular or granular layers of the cerebellar cortex. It is proposed that a decrease in GABA receptors in the dentate nucleus results in disinhibition of cerebellar pacemaker output activity, propagating along the cerebello-thalamo-cortical pathways to generate tremors. Correction of such defective cerebellar GABAergic drive could have a therapeutic effect in essential tremor.

Synaptic action of the olivocerebellar system on cerebellar nuclear spike activity.

Cerebellar output is necessary for the ideal implementation of many nervous system functions, particularly motor coordination. A key step toward understanding the generation of this output is characterizing the factors that shape the activity of the cerebellar nuclei (CN). There are four major sources of synaptic input that modulate CN activity; collaterals of climbing and mossy fibers are two, and the remaining two are provided by Purkinje cell (PC) axons in the form of simple spikes (SSs) and complex spikes (CSs). Most hypotheses of cerebellar function focus on SSs as the primary determinant of CN activity. However, it is likely that CSs also cause significant direct effects on CN activity, something that is rarely considered. To explore this possibility, we recorded from synaptically connected PC-CN neuron cell pairs in rats. Cross-correlograms of CS and CN activity from such recordings demonstrate that spontaneous CSs have a strong inhibitory effect on CN activity, apparently sufficient, in some cases, to trigger changes in the intrinsic excitability of the CN neuron that long outlast the underlying CS-mediated GABAergic IPSP. Furthermore, many CS-CN correlograms show an initial excitatory response, demonstrating the ability of climbing fiber collaterals to significantly excite CN neurons. A substantial fraction (24%) of correlograms displayed an excitation-inhibition sequence, providing evidence that a CN neuron often receives collaterals from the same olivocerebellar axons as innervate the PCs projecting to it. Thus, excitation followed by inhibition appears to be a hard-wired response pattern of many CN neurons to olivocerebellar activity.

Neural circuitry and plasticity mechanisms underlying delay eyeblink conditioning

Pavlovian eyeblink conditioning has been used extensively as a model system for examining the neural mechanisms underlying associative learning. Delay eyeblink conditioning depends on the intermediate cerebellum ipsilateral to the conditioned eye. Evidence favors a two-site plasticity model within the cerebellum with long-term depression of parallel fiber synapses on Purkinje cells and long-term potentiation of mossy fiber synapses on neurons in the anterior interpositus nucleus. Conditioned stimulus and unconditioned stimulus inputs arise from the pontine nuclei and inferior olive, respectively, converging in the cerebellar cortex and deep nuclei. Projections from subcortical sensory nuclei to the pontine nuclei that are necessary for eyeblink conditioning are beginning to be identified, and recent studies indicate that there are dynamic interactions between sensory thalamic nuclei and the cerebellum during eyeblink conditioning. Cerebellar output is projected to the magnocellular red nucleus and then to the motor nuclei that generate the blink response(s). Tremendous progress has been made toward determining the neural mechanisms of delay eyeblink conditioning but there are still significant gaps in our understanding of the necessary neural circuitry and plasticity mechanisms underlying cerebellar learning.

Evaluating the adaptive-filter model of the cerebellum.

The adaptive-filter model of the cerebellar microcircuit is in widespread use, combining as it does an explanation of key microcircuit features with well-specified computational power. Here we consider two methods for its evaluation. One is to test its predictions concerning relations between cerebellar inputs and outputs. Where the relevant experimental data are available, e.g. for the floccular role in image stabilization, the predictions appear to be upheld. However, for the majority of cerebellar microzones these data have yet to be obtained. The second method is to test model predictions about details of the microcircuit. We focus on features apparently incompatible with the model, in particular non-linear patterns in Purkinje cell simple-spike firing. Analysis of these patterns suggests the following three conclusions. (i) It is important to establish whether they can be observed during task-related behaviour. (ii) Highly non-linear models based on these patterns are unlikely to be universal, because they would be incompatible with the (approximately) linear nature of floccular function. (iii) The control tasks for which these models are computationally suited need to be identified. At present, therefore, the adaptive filter remains a candidate model of at least some cerebellar microzones, and its evaluation suggests promising lines for future enquiry.

The mysterious microcircuitry of the cerebellar nuclei.

The microcircuitry of cerebellar cortex and, in particular, the physiology of its main element, the Purkinje neuron, has been extensively investigated and described. However, activity in Purkinje neurons, either as single cells or populations, does not directly mediate the cerebellar effects on the motor effector systems. Rather, the result of the entire cerebellar cortical computation is passed to the relatively small cerebellar nuclei that act as the final, integrative processing unit in the cerebellar circuitry. The nuclei ultimately control the temporal and spatial features of the cerebellar output. Given this key role, it is striking that the internal organization and the connectivity with afferent and efferent pathways in the cerebellar nuclei are rather poorly known. In the present review, we discuss some of the many critical shortcomings in the understanding of cerebellar nuclei microcircuitry: the extent of convergence and divergence of the cerebellar cortical pathway to the various cerebellar nuclei neurons and subareas, the possible (lack of) conservation of the finely-divided topographical organization in the cerebellar cortex at the level of the nuclei, as well as the absence of knowledge of the synaptic circuitry within the cerebellar nuclei. All these issues are important for predicting the pattern-extraction and encoding capabilities of the cerebellar nuclei and, until resolved, theories and models of cerebellar motor control and learning may err considerably.

Topography of Cerebellar Deficits in Humans

The cerebellum is a key-piece for information processing and is involved in numerous motor and nonmotor activities, thanks to the anatomical characteristics of the circuitry, the enormous computational capabilities and the high connectivity to other brain areas. Despite its uniform cytoarchitecture, cerebellar circuitry is segregated into functional zones. This functional parcellation is driven by the connectivity and the anatomo-functional heterogeneity of the numerous extra-cerebellar structures linked to the cerebellum, principally brain cortices, precerebellar nuclei and spinal cord. Major insights into cerebellar functions have been gained with a detailed analysis of the cerebellar outputs, with the evidence that fundamental aspects of cerebrocerebellar operations are the closed-loop circuit and the predictions of future states. Cerebellar diseases result in disturbances of accuracy of movements and lack of coordination. The cerebellar syndrome includes combinations of oculomotor disturbances, dysarthria and other speech deficits, ataxia of limbs, ataxia of stance and gait, as well as often more subtle cognitive/behavioral impairments. Our understanding of the corresponding anatomo-functional maps for the human cerebellum is continuously improving. We summarize the topography of the clinical deficits observed in cerebellar patients and the growing evidence of a regional subdivision into motor, sensory, sensorimotor, cognitive and affective domains. The recently described topographic dichotomy motor versus nonmotor cerebellum based upon anatomical, functional and neuropsychological studies is also discussed.

Human Locomotor Adaptive Learning Is Proportional to Depression of Cerebellar Excitability

Human locomotor adaptive learning is thought to involve the cerebellum, but the neurophysiological mechanisms underlying this process are not known. While animal research has pointed to depressive modulation of cerebellar outputs, a direct correlation between adaptive learning and cerebellar depression has never been demonstrated. Here, we used transcranial magnetic stimulation to assess excitability changes occurring in the cerebellum and primary motor cortex (M1) after individuals learned a new locomotor pattern on a split-belt treadmill. To control for potential changes associated to task performance complexity, the same group of subjects was also assessed after performing 2 other locomotor tasks that did not elicit learning. We found that only adaptive learning resulted in reduction of cerebellar inhibition. This effect was strongly correlated with the magnitude of learning (r = 0.78). In contrast, M1 excitability changes were not specific to learning but rather occurred in association with task complexity performance. Our results demonstrate that locomotor adaptive learning in humans is proportional to cerebellar excitability depression. This finding supports the theory that adaptive learning is mediated, at least in part, by long-term depression in Purkinje cells. This knowledge opens the opportunity to target cerebellar processes with noninvasive brain stimulation to enhance motor learning.

Cerebellar cortical output encodes temporal aspects of rhythmic licking movements and is necessary for normal licking frequency

Rodents consume water by performing stereotypic, rhythmic licking movements that are believed to be controlled by brainstem pattern-generating circuits. Previous work has shown that synchronized population activity of inferior olive neurons was phase-locked to the licking rhythm in rats, suggesting a cerebellar involvement in temporal aspects of licking behavior. However, what role the cerebellum has in licking behavior and whether licking is represented in the high-frequency simple spike output of Purkinje cells remains unknown. We recorded Purkinje cell simple and complex spike activity in awake mice during licking, and determined the behavioral consequences of loss of cerebellar function. Mouse cerebellar cortex contained a multifaceted representation of licking behavior encoded in the simple spike activities of Purkinje cells distributed across Crus I, Crus II and lobus simplex of the right cerebellar hemisphere. Lick-related Purkinje cell simple spike activity was modulated rhythmically, phase-locked to the lick rhythm, or non-rhythmically. A subpopulation of lick-related Purkinje cells differentially represented lick interval duration in their simple spike activity. Surgical removal of the cerebellum or temporary pharmacological inactivation of the cerebellar nuclei significantly slowed the licking frequency. Fluid licking was also less efficient in mice with impaired cerebellar function, indicated by a significant decline in the volume per lick fluid intake. The gross licking movement appeared unaffected. Our results suggest a cerebellar role in modulating the frequency of the central pattern-generating circuits controlling fluid licking and in the fine coordination of licking, while contributing little to the coordination of the gross licking movement.