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Here we investigate stimulus generalization in a cerebellar learning paradigm, called eyeblink conditioning. Mice were conditioned to close their eyes in response to a 10 kHz tone by repeatedly pairing this tone with an air puff to the eye 250 ms after tone onset. After ten consecutive days of training, when mice showed reliable conditioned eyelid responses to the 10 kHz tone, we started to expose them to tones with other frequencies, ranging from 2 to 20 kHz. We found that mice had a strong generalization gradient, whereby the probability and amplitude of conditioned eyelid responses gradually decreases depending on the dissimilarity with the 10 kHz tone. Tones with frequencies closest to 10kHz evoked the most and largest conditioned eyelid responses and each step away from the 10 kHz tone resulted in fewer and smaller conditioned responses. In addition, we found that tones with lower frequencies resulted in conditioned responses that peaked earlier after tone onset compared to those to tones with higher frequencies. Together, our data show prominent generalization patterns in cerebellar learning. Since the known function of cerebellum is rapidly expanding from pure motor control to domains that include cognition, reward-learning, fear-learning, social function, and even addiction, our data implies generalization controlled by cerebellum in all these domains.Significance StatementConditioned stimuli that resemble each other will evoke a rather similar behavioral response. Here, we investigate this phenomenon of stimulus generalization using Pavlovian eyeblink conditioning to probe cerebellar function. Cerebellum is a brain structure whose known function is rapidly expanding from pure motor control to domains that include cognition, reward-learning, fear-learning, social function, and even addiction. Since we found a strong generalization of the auditory stimulus in eyeblink conditioning, our data implies an important role for generalization in motor and non-motor domains.

BACKGROUND Onco-neurosurgical practice still relies heavily on pre-operatively acquired images to guide intra-operative decision-making for safe tumor removal, a practice with inherent pitfalls such as registration inaccuracy due to brain shift, and lack of real-time (functional) feedback. Exploiting the opportunity for real-time imaging of the exposed brain can improve intra-operative decision-making, neurosurgical safety and patient outcomes. Previously, we described functional Ultrasound (fUS) as a high-resolution, depth-resolved imaging technique able to detect functional regions and vascular morphology during awake resections. Here, we present for the first time fUS as a fully integrated, MRI/CT-registered imaging modality in the OR. MATERIAL AND METHODS fUS relies on high-frame-rate (HFR) ultrasound, making the technique sensitive for very small motions caused by vascular dynamics (µDoppler) and allowing measurements of changes in cerebral blood volume (CBV) with micrometer-millisecond precision. This opens up the possibility to 1) detect functional response, as CBV-changes reflect changes in metabolism of activated neurons through neurovascular coupling and 2) visualize in-vivo vascular morphology of tumor and healthy tissue. During a range of anesthetized and awake onco-neurosurgical procedures we acquired images of brain and spinal cord using conventional linear ultrasound probes connected to an experimental acquisition unit. Building on Brainlab’s ‘Cranial Navigation’ and ‘Intra-Operative Ultrasound’ modules, we could co-register our intra-operative Power Doppler Images (PDIs) to patient-registered MRI/CT-data. Using the ‘IGTLink’ research interface, we could access and store real-time tracking data for informed volume reconstructions in post-processing. RESULTS Intra-operative fUS could be registered to MRI/CT-images in real-time, showing overlays of PDIs at imaging depths of >5 centimeters. During meningioma resections, these co-registered PDIs revealed fUS’ ability to visualize the tumor’s feeding vessels and surrounding healthy vasculature prior to durotomy, with a level of detail unprecedented by conventional MRI-sequences. Comparing post-operatively reconstructed 3D-vascular maps of pre- and post-durotomy acquisitions, further confirmed the dural dependency of the vascular network feeding the tumor. During awake resections, fUS revealed distinct functional areas as activated during motor and language tasks. CONCLUSION fUS is a new real-time, high-resolution and depth-resolved imaging technique, combining characteristics uniquely beneficial for a potential image-guided resection tool. The successful integration of fUS in the onco-neurosurgical OR demonstrated by our team, is an essential step towards clinical integration of fUS, as well as the technique’s validation against modalities such as MRI and CT.

Mathematical models with varying degrees of complexity have been proposed and simulated in an attempt to represent the intricate mechanisms of the human neuron. One of the most biochemically realistic and analytical models, based on the Hodgkin–Huxley (HH) model, has been selected for study in this paper. In order to satisfy the model’s computational demands, we present a simulator implemented on Intel Xeon Phi Knights Landing manycore processors. This high-performance platform features an x86-based architecture, allowing our implementation to be portable to other common manycore processing machines. This is reinforced by the fact that Phi adopts the popular OpenMP and MPI programming models. The simulator performance is evaluated when calculating neuronal networks of varying sizes, density and network connectivity maps. The evaluation leads to an analysis of the neuronal synaptic patterns and their impact on performance when tackling this type of workload on a multinode system. It will be shown that the simulator can calculate 100 ms of simulated brain activity for up to 2 millions of biophysically-accurate neurons and 2 billion neuronal synapses within one minute of execution time. This level of performance renders the application an efficient solution for large-scale detailed model simulation.

The eyeblink conditioning paradigm captures an elementary form of associative learning in a neural circuitry that is understood to an extraordinary degree. Cerebellar cortical Purkinje cell simple spike suppression is widely regarded as the main process underlying conditioned responses (CRs), leading to disinhibition of neurons in the cerebellar nuclei that innervate eyelid muscles downstream. However, recent work highlights the addition of a conditioned Purkinje cell complex spike response, which at the level of the interposed nucleus seems to translate to a transient spike suppression that can be followed by a rapid spike facilitation. Here, we review the characteristics of these responses at the cerebellar cortical and nuclear level, and discuss possible origins and functions.

Functional ultrasound (fUS) is a relatively new imaging modality to study the brain with a high spatiotemporal resolution and a wide field-of-view. In fUS detailed images of cerebral blood flow and volume are used to derive functional information, as changes in local flow and/or volume may reflect neuronal activation through neurovascular coupling. Most fUS studies so far have been performed in rats. Translating fUS to mice, which is a favorable animal model for neuroscience, pleads for a higher spatial resolution than what has been reported so far. As a consequence the temporal sampling of the blood flow should also be increased in order to adequately capture the wide range in blood velocities, as the Doppler shifts are inversely proportional to the spatial resolution. Here we present our first detailed images of the mouse brain vasculature at high spatiotemporal resolution. In addition we show some early experimental work on tracking brain activity upon local electrical stimulation.

Plastic changes in the efficacy of synapses are widely regarded to represent mechanisms underlying memory formation. So far, evidence for learning-dependent, new neuronal wiring is limited. In this study, we demonstrate that pavlovian eyeblink conditioning in adult mice can induce robust axonal growth and synapse formation in the cerebellar nuclei. Thisde novowiring is both condition specific and region specific because it does not occur in pseudoconditioned animals and is particularly observed in those parts of the cerebellar nuclei that have been implicated to be involved in this form of motor learning. Moreover, the number of new mossy fiber varicosities in these parts of the cerebellar nuclei is positively correlated with the amplitude of conditioned eyelid responses. These results indicate that outgrowth of axons and concomitant occurrence of new terminals may, in addition to plasticity of synaptic efficacy, contribute to the formation of memory.

During delay classical conditioning of the eyelid response, which models both associative learning and motor learning, the essential memory is formed in the cerebellum. In this chapter we discuss how the cerebellar cortex and cerebellar nuclei may synergistically interact to generate conditioned eyelid responses. For this, we first describe some important behavioral aspects of eyeblink conditioning, including new data on the effects of conditional stimulus (CS) durations that are either shorter or longer than the trained CS–unconditional stimulus interval. Second, we provide a systematic overview of the neuroanatomical circuits involved in eyeblink conditioning. In the third part we discuss the contributions of various forms of neural plasticity in the cerebellar cortex and cerebellar nuclei to the formation of a memory trace during eyeblink conditioning. Together, these parts allow us to synthesize an overview of putative encoding mechanisms underlying eyeblink conditioning.

Fragile X syndrome (FXS) is the most common inherited form of intellectual disability. Patients with FXS do not only suffer from cognitive problems, but also from abnormalities/deficits in procedural memory formation. It has been proposed that a lack of fragile X mental retardation protein (FMRP) leads to altered long-term plasticity by deregulation of various translational processes at the synapses, and that part of these impairments might be rescued by the inhibition of type I metabotropic glutamate receptors (mGluRs). We recently developed the Erasmus Ladder, which allows us to test, without any invasive approaches, simultaneously, both procedural memory formation and avoidance behavior during unperturbed and perturbed locomotion in mice. Here, we investigated the impact of a potent and selective mGluR5 inhibitor (Fenobam) on the behavior of Fmr1 KO mice during the Erasmus Ladder task. Fmr1 KO mice showed deficits in associative motor learning as well as avoidance behavior, both of which were rescued by intraperitoneal administration of Fenobam. While the Fmr1 KO mice did benefit from the treatment, control littermates suffered from a significant negative side effect in that their motor learning skills, but not their avoidance behavior, were significantly affected. On the basis of these studies in the FXS animal model, it may be worthwhile to investigate the effects of mGluR inhibitors on both the cognitive functions and procedural skills in FXS patients. However, the use of mGluR inhibitors appears to be strongly contraindicated in healthy controls or non-FXS patients with intellectual disability.

Neurofibromatosis type 1 (NF1) is an autosomal dominantly inherited disease, characterized by various neurocutaneous symptoms, cognitive impairments and problems in fine and gross motor performance. Although cognitive deficits in NF1 have been attributed to increased release of the inhibitory neurotransmitter γ-amino butyric acid (GABA) in the hippocampus, the origin of the motor deficits is unknown. Cerebellar Purkinje cells, the sole output neurons of the cerebellar cortex, are GABAergic neurons and express neurofibromin at high levels, suggesting an important role for the cerebellum in the observed motor deficits in NF1. To test this, we determined the cerebellar contribution to motor problems in Nf1(+/-) mice, a validated mouse model for NF1. Using the Rotarod, a non-specific motor performance test, we confirmed that, like NF1 patients, Nf1(+/-) mice have motor deficits. Next, to evaluate the role of the cerebellum in these deficits, mice were subjected to cerebellum-specific motor performance and learning tests. Nf1(+/-) mice showed no impairment on the Erasmus ladder, as step time and number of missteps were not different. Furthermore, when compensatory eye movements were tested, no performance deficits were found in the optokinetic reflex and vestibulo-ocular reflex in the dark (VOR) or in the light (VVOR). Finally, Nf1(+/-) mice successfully completed short- and long-term VOR adaptation paradigms, tests that both depend on cerebellar function. Thus, despite the confirmed presence of motor performance problems in Nf1(+/-) mice, we found no indication of a cerebellar component. These results, combined with recent clinical data, suggest that cerebellar function is not overtly affected in NF1 patients.

SummaryCerebellar motor learning is required to obtain procedural skills. Studies have provided supportive evidence for a potential role of kinase-mediated long-term depression (LTD) at the parallel fiber to Purkinje cell synapse in cerebellar learning. Recently, phosphatases have been implicated in the induction of potentiation of Purkinje cell activities in vitro, but it remains to be shown whether and how phosphatase-mediated potentiation contributes to motor learning. Here, we investigated its possible role by creating and testing a Purkinje cell-specific knockout of calcium/calmodulin-activated protein-phosphatase-2B (L7-PP2B). The selective deletion of PP2B indeed abolished postsynaptic long-term potentiation in Purkinje cells and their ability to increase their excitability, whereas LTD was unaffected. The mutants showed impaired “gain-decrease” and “gain-increase” adaptation of their vestibulo-ocular reflex (VOR) as well as impaired acquisition of classical delay conditioning of their eyeblink response. Thus, our data indicate that PP2B may indeed mediate potentiation in Purkinje cells and contribute prominently to cerebellar motor learning.Video Abstract

The brain generates many rhythmic activities, and the olivo-cerebellar system is not an exception. In recent years, the cerebellum has revealed activities ranging from low frequency to very high-frequency oscillations. These rhythms depend on the brain functional state and are typical of certain circuit sections or specific neurons. Interestingly, the granular layer, which gates sensorimotor and cognitive signals to the cerebellar cortex, can also sustain low frequency (7-25 Hz) and perhaps higher-frequency oscillations. In this review we have considered (i) how these oscillations are generated in the granular layer network depending on intrinsic electroresponsiveness and circuit connections, (ii) how these oscillations are correlated with those in other cerebellar circuit sections, and (iii) how the oscillating cerebellum communicates with extracerebellar structures. It is suggested that the granular layer can generate oscillations that integrate well with those generated in the inferior olive, in deep-cerebellar nuclei and in Purkinje cells. These rhythms, in turn, might play a role in cognition and memory consolidation by interacting with the mechanisms of long-term synaptic plasticity. (C) 2009 IBRO. Published by Elsevier Ltd. All rights reserved.

Stopping a clinical trial without reaching the final objective is not the ideal outcome any researcher wants; sometimes ceasing is inevitable. Due to marginal inclusion of patients we were forced to cease our randomized clinical trial on the effectiveness of proprioceptive training on the development of chronic whiplash complaints a year after the start. Although incidence figures demonstrate that recruitment of the planned number of whiplash patients would be easily feasible, we were unable to enroll the amount of subjects. Several motives can be proposed that would have prevented this obliged halting from happening. Other studies also report impracticability of the planned number of whiplash injury patients. (c) 2008 Elsevier Inc. All rights reserved.

The climbing fibre (CF) input controls cerebellar Purkinje cell (PC) activity as well as synaptic plasticity at parallel fibre (PF)-PC synapses. Under high activity conditions, CFs release not only glutamate, but also the neuropeptide corticotropin-releasing factor (CRF). Brief periods of such high CF activity can lead to the induction of long-term depression (LTD) at CF-PC synapses. Thus, we have examined for the first time the role of CRF in regulating excitatory postsynaptic currents (EPSCs) and long-term plasticity at this synapse. Exogenous application of CRF alone transiently mimicked three aspects of CF-LTD, causing reductions in the CF-evoked excitatory postsynaptic current, complex spike second component and complex spike afterhyperpolarization. The complex spike first component is unaffected by CF-LTD induction and was similarly unaffected by CRF. Application of a CRF receptor antagonist reduced the expression amplitude and induction probability of CF-LTD monitored at the EPSC level. Collectively, these results suggest that under particular sensorimotor conditions, co-release of CRF from climbing fibres could down-regulate excitatory transmission and facilitate LTD induction at CF-PC synapses. Inhibition of either protein kinase C (PKC) or protein kinase A (PKA) attenuated the effects of CRF upon CF-EPSCs. We have previously shown that CF-LTD induction is PKC-dependent, and here demonstrate PKA-dependence as well. These results suggest that both the acute effects of CRF on CF-EPSCs as well as the facilitating effect of CRF on CF-LTD induction can be explained by a CRF-mediated recruitment of PKC and PKA.

Williams Syndrome (WS, [MIM 194050]) is a disorder caused by a hemizygous deletion of 25-30 genes on chromosome 7q11.23. Several of these genes including those encoding cytoplasmic linker protein-115 (CYLN2) and general transcription factors (GTF2I and GTF2IRD1) are expressed in the brain and may contribute to the distinct neurological and cognitive deficits in WS patients. Recent studies of patients with partial deletions indicate that hemizygosity of GTF2I probably contributes to mental retardation in WS. Here we investigate whether CYLN2 and GTF2IRD1 contribute to the motoric and cognitive deficits in WS. Behavioral assessment of a new patient in which STX1A and LIMK1, but not CYLN2 and GTF2IRD1, are deleted showed that his cognitive and motor coordination functions were significantly better than in typical WS patients. Comparative analyses of gene specific CYLN2 and GTF2IRD1 knockout mice showed that a reduced size of the corpus callosum as well as deficits in motor coordination and hippocampal memory formation may be attributed to a deletion of CYLN2, while increased ventricle volume can be attributed to both CYLN2 and GTF2IRD1. We conclude that the motor and cognitive deficits in Williams Syndrome are caused by a variety of genes and that heterozygous deletion of CYLN2 is one of the major causes responsible for such dysfunctions.

Echinoderm microtubule-associated protein (EMAP) is the major microtubule binding protein in dividing sea urchin (Strongylocentrotus purpuratus) eggs. Echinoderm microtubule-associated protein like protein 4 (Eml4, restrictedly overexpressed proliferation-associated protein 120 kDa (Ropp120)) is one of the five mammalian EMAP homologues, the cellular function of which remains to be elucidated. In our first set of experiments we determined the spatio-temporal expression pattern of Eml4 in mouse brain. Our results demonstrate that Eml4 is a highly developmentally regulated gene with high expression levels in the developing nervous system of E11 embryos declining to low levels in adult. Spatially, Eml4 expression becomes restricted to the olfactory bulb, hippocampus and cerebellum. Transient transfection of a fusion construct of full-length mouse Eml4 with green fluorescent protein (GFP-Eml4) into Cos7 and HeLa cells resulted in colocalization of GFP-Eml4 with microtubules. This colocalization was observed both with microtubules of non-dividing cells and with the mitotic spindle of dividing cells. In addition, transient overexpression of GFP-Eml4 in Cos7 cells resulted in microtubules that were resistant to nocodazole treatment suggesting that Eml4 stabilizes microtubules. A consequence of microtubule stabilization is a net reduction in the amount of free tubulin. Microtubule stabilizing proteins therefore are expected to indirectly decrease the microtubule growth rate. Indeed, transient transfection of GFP-Eml4 resulted in a marked decrease in the microtubule growth rate, which is in line with our hypothesis that Eml4 functions as a microtubule stabilizing protein. In summary, our results suggest that Eml4 is a developmentally regulated protein that colocalizes with and stabilizes microtubules.

Vestibular paradigms are widely used for investigating mechanisms underlying cerebellar motor learning. These include adaptation of the vestibuloocular reflex (VOR) after visual-vestibular mismatch training and vestibular compensation after unilateral damage to the vestibular apparatus. To date, various studies have shown that VOR adaptation may be supported by long-term depression (LTD) at the parallel fiber to Purkinje cell synapse. Yet it is unknown to what extent vestibular compensation may depend on this cellular process. Here we investigated adaptive gain changes in the VOR and optokinetic reflex during vestibular compensation in transgenic mice in which LTD is specifically blocked in Purkinje cells via expression of a peptide inhibitor of protein kinase C (L7-PKCi mutants). The results demonstrate that neither the strength nor the time course of vestibular compensation are affected by the absence of LTD. In contrast, analysis of vestibular compensation in spontaneous mutants that lack a functional olivo-cerebellar circuit ( lurchers) shows that this form of motor learning is severely impaired. We conclude that oculomotor plasticity during vestibular compensation depends critically on intact cerebellar circuitry but not on the occurrence of cerebellar LTD.

This paper presents the design and implementation of a real-time epilepsy detection filter that is suitable for closed-loop seizure suppression. The design aims to minimize the detection delay, while a reasonable average detection rate is obtained. The filter is based on a complex Morlet wavelet and uses an adaptive thresholding strategy for the seizure discrimination. This relatively simple configuration allows the algorithm to run on a cheap and readily available microprocessor prototyping platform. The performance of the filter is verified using both in vivo real-time measurements as well as simulations over a pre-recorded EEG dataset (29.75 hours with 1914 seizures). An average detection delay of 492 ms is achieved with a sensitivity of 96.03% and a specificity of 93.60%.

Summary Absence of functional FMRP causes Fragile X syndrome. Abnormalities in synaptic processes in the cerebral cortex and hippocampus contribute to cognitive deficits in Fragile X patients. So far, the potential roles of cerebellar deficits have not been investigated. Here, we demonstrate that both global and Purkinje cell-specific knockouts of Fmr1 show deficits in classical delay eyeblink conditioning in that the percentage of conditioned responses as well as their peak amplitude and peak velocity are reduced. Purkinje cells of these mice show elongated spines and enhanced LTD induction at the parallel fiber synapses that innervate these spines. Moreover, Fragile X patients display the same cerebellar deficits in eyeblink conditioning as the mutant mice. These data indicate that a lack of FMRP leads to cerebellar deficits at both the cellular and behavioral levels and raise the possibility that cerebellar dysfunctions can contribute to motor learning deficits in Fragile X patients.

Contains fulltext : 48072.pdf (Publisher’s version ) (Closed access) While firing rate is well established as a relevant parameter for encoding information exchanged between neurons, the significance of other parameters is more conjectural. Here, we show that regularity of neuronal spike activities affects sensorimotor processing in tottering mutants, which suffer from a mutation in P/Q-type voltage-gated calcium channels. While the modulation amplitude of the simple spike firing rate of their floccular Purkinje cells during optokinetic stimulation is indistinguishable from that of wild-types, the regularity of their firing is markedly disrupted. The gain and phase values of tottering's compensatory eye movements are indistinguishable from those of flocculectomized wild-types or from totterings with the flocculus treated with P/Q-type calcium channel blockers. Moreover, normal eye movements can be evoked in tottering when the flocculus is electrically stimulated with regular spike trains mimicking the firing pattern of normal simple spikes. This study demonstrates the importance of regularity of firing in Purkinje cells for neuronal information processing.

Whiplash-associated disorders (WAD) are a major problem in the Western world, which put a formidable financial burden on modern society and which evoke an emerging debate on the true nature of their origin. To date there is no generally accepted test that allows us to diagnose WAD objectively. Because whiplash injury causes dysfunction of proprioception in the neck, we investigated the characteristics of the cervico-ocular reflex (COR) of presumptive WAD patients. These patients and age-matched healthy controls were rotated at different stimulus peak velocities in the dark while their head was fixed in space. The gain values of the COR were significantly increased in the patient population at a wide range of stimulus peak velocities with maximum difference at the lower frequencies (p = 0.037, ANOVA). Hence, although larger numbers of patients should be measured, the COR gain appears to be a parameter that may permit an objective diagnosis of WAD.

The cervico-ocular reflex (COR) works in conjunction with the vestibulo-ocular reflex (VOR) and the optokinetic reflex (OKR) in order to prevent visual slip over the retina during head movement. The COR induces eye movements in response to proprioceptive signals from the neck. We investigated whether the COR gain can be adapted by inducing a mismatch between vision and neck proprioception, in analogy to VOR adaptation. Thirteen healthy subjects were rotated in the dark in a trunk-to-head manner (the head fixed in space while the body passively rotated sinusoidally with a peak velocity of 1.25 degrees /s). Eye movements were recorded with infrared video-oculography under various adaptive conditions. Analysis showed a small but significant reduction in COR gain in the suppression conditions. This means that the cervico-ocular reflex can be modified after only 10 min of concurrent visual and cervical stimulation.

Contains fulltext : 57891.pdf (Publisher’s version ) (Closed access) Cerebellar long-term depression (LTD) at parallel fibre-Purkinje cell (P-cell) synapses is thought to embody neuronal information storage for motor learning. Transgenic L7-protein kinase C inhibitor (PKCI) mice in which cerebellar LTD is selectively blocked do indeed exhibit impaired adaptation in the vestibulo-ocular reflex (VOR) while their default oculomotor performance is unaffected. Although supportive, these data do not definitively establish a causal link between memory storage required for motor learning and cerebellar LTD. As the L7-PKCI transgene is probably activated from the early stages of P-cell development, an alternative could be that P-cells develop abnormal signals in L7-PKCI mutants, disturbing mechanisms of motor learning that rely on proper P-cell outputs. To test this alternative hypothesis, we studied simple spike (SS) and complex spike (CS) activity of vertical axis P-cells in the flocculus of L7-PKCI mice and their wild-type littermates during sinusoidal optokinetic stimulation. Both SS and CS discharge dynamics appeared to be very similar in wild-type and transgenic P-cells at all stimulus frequencies (0.05-0.8 Hz). The CS activity of all vertical axis cells increased with contralateral stimulus rotation and lagged ipsiversive eye velocity by 165-180 degrees. The SS modulation was roughly reciprocal to the CS modulation and lagged ipsiversive eye velocity by approximately 15 degrees. The baseline SS and CS discharge characteristics were indistinguishable between the two genotypes. We conclude that the impaired VOR learning in L7-PKCI mutants does not reflect fundamental aberrations of the cerebellar circuitry. The data thus strengthen the evidence that cerebellar LTD is implicated in rapid VOR learning but not in the development of normal default response patterns.

We have isolated a novel transcript with homology to the major microtubule-associated protein in dividing sea urchin embryos, EMAP. The protein has a predicted MW of ∼180 kDa and we have named it Eml5 (EMAP-like protein 5, GenBank accession no. AY445136). Eml5 contains 11 putative WD40 domains and 3 hydrophobic stretches of 43 aa, HELP domains, which have been suggested to be involved in microtubule binding. Eml5 appears to consist of two tandem repeats of the complete EMAP protein separated by a putative dimerization domain. Eml5 mRNA and protein is expressed at high levels in the hippocampus, cerebellum and olfactory bulb, as determined by in situ hybridization and immunocytochemistry. Eml5 transcripts can be detected in fore- and hindbrain structures from embryonic day 13 onwards. Because other EMAP-like proteins are involved in regulating microtubule dynamics, it is likely that Eml5 plays a role in the regulation of cytoskeletal rearrangements during neuronal development and in adult brain

The induction of cerebellar long-term depression (LTD) at the parallel fibre-Purkinje cell synapse is selectively blocked in L7-PKCi transgenic mice, rendering these mice unable to adaptively modify their vestibulo-ocular reflex (VOR) during visuo-vestibular training for a few hours. Despite this deficit, their eye movement performance as well as their general motor behaviour appears unaffected. This combination suggests that, in the long term, residual forms of plasticity in the vestibulo-cerebellar circuitry can compensate for the absence of cerebellar LTD. To investigate whether LTD-deficient mice exhibit motor learning in the long run, we subjected L7-PKCi transgenic mice to visuo-vestibular training paradigms that were aimed at either increasing or decreasing the VOR response in the course of eight consecutive days. During the increasing paradigm, the VOR gain of transgenic mice increased significantly, while VOR gain decreased and VOR phase-lead increased during the decreasing paradigm. The impact of these long training periods on the VOR was significantly smaller in LTD-deficient mice than in wild type littermates. Thus, while LTD may be necessary for short-term VOR adaptation, it facilitates but is not required for long-term adaptation of the VOR.

Electrotonic coupling by gap junctions between neurons in the inferior olive has been claimed to underly complex spike (CS) synchrony of Purkinje cells in the cerebellar cortex and thereby to play a role in the coordination of movements. Here, we investigated the motor performance of mice that lack connexin36 (Cx36), which appears necessary for functional olivary gap junctions. Cx36 null-mutants are not ataxic, they show a normal performance on the accelerating rotorod, and they have a regular walking pattern. In addition, they show normal compensatory eye movements during sinusoidal visual and/or vestibular stimulation. To find out whether the normal motor performance in mutants reflects normal CS activity or some compensatory mechanism downstream of the cerebellar cortex, we determined the CS firing rate, climbing-fiber pause, and degree of CS synchrony. None of these parameters in the mutants differed from those in wildtype littermates. Finally, we investigated whether the role of coupling becomes apparent under challenging conditions, such as during application of the tremorgenic drug harmaline, which specifically turns olivary neurons into an oscillatory state at a high frequency. In both the mutants and wildtypes this application induced tremors of a similar duration with similar peak frequencies and amplitudes. Thus surprisingly, the present data does not support the notion that electrotonic coupling by gap junctions underlies synchronization of olivary spike activity and that these gap junctions are essential for normal motor performance.

The mouse connexin 36 (Cx36) gene was mapped on chromosome 2 and an identical transcriptional start site was determined in brain and retina on exon I. Rabbit polyclonal antibodies to the presumptive cytoplasmic loop of the Cx36 protein recognized in immunohistochemical analyses Cx36 expression in the retina, olfactory bulb, hippocampus, inferior olive and cerebellum. In olivary neurons strong punctate labeling at dendritic cell contacts and weaker labeling in the cytoplasm of dendrites were shown by immuno electron microscopy. After expression of mouse Cx36 cDNA in human HeLa cells, neurobiotin transfer was increased 1.8-fold and electrical conductance at least 15-fold compared to untransfected HeLa cells. No Lucifer Yellow transfer was detected in either untransfected or Cx36 transfected HeLa cells. Single Cx36 channels in transfected HeLa cells showed a unitary conductance of 14.3 + or - 0. 8 pS. The sensitivity of Cx36 channels to transjunctional voltage was low in both HeLa-Cx36 cells and Xenopus oocytes expressing mouse Cx36. No increased transfer of neurobiotin was detected in heterotypic gap junctions formed by Cx36 and 9 other connexins expressed in HeLa cells. Our results suggest that Cx36 channels function as electrical synapses for transmission of electrical and metabolic signals between neurons in the central nervous system.

Apoptosis is a critical cellular event during several stages of neuronal development. Recently, we have shown that biotinylated annexin V detects apoptosis in vivo in various cell lineages of a wide range of species by binding to phosphatidylserines that are exposed at the outer leaflet of the plasma membrane. In the present study, we tested the specificity by which annexin V binds apoptotic neurons, and subsequently investigated developmental cell death in the central and peripheral nervous system of early mouse embryos at both the cellular and histological level, and compared the phagocytic clearance of apoptotic neurons with that of apoptotic mesodermal cells. Our data indicate: (i) that biotinylated annexin V can be used as a sensitive marker that detects apoptotic neurons, including their extensions at an early stage during development; (ii) that apoptosis plays an important part during early morphogenesis of the central nervous system, and during early quantitative matching of brain-derived neurotrophic factor and neurotrophic factor 3 responsive postmitotic large clear neurons in the peripheral ganglia with their projection areas; and (iii) that apoptotic neurons are removed by a process that differs from classical phagocytosis of non-neuronal tissues.

Two different populations of projection neurons are intermingled in the cerebellar nuclei. One group consists of small, γ-aminobutyric acid-containing (GABAergic) neurons that project to the inferior olive, and the other group consists of larger, non-GABAergic neurons that provide an input to one or more, usually premotor, centers in the brainstem, such as the red nucleus, the thalamus, and the superior colliculus. All cerebellar nuclear neurons are innervated by GABAergic Purkinje cells. In this study, we investigated whether individual Purkinje cells of the C1 zone of the paramedian lobe of the rat innervate both groups of projection neurons in the anterior interposed nucleus. Two different, retrogradely transported tracers, either cholera toxin β subunit (CTb) or wheat germ agglutinin coupled to horseradish peroxidase (WGA-HRP) and a gold lectin tracer were injected into the red nucleus and the inferior olive, respectively, whereas Purkinje cell axons were anterogradely labeled with biotinylated dextran amine (BDA) injected into the paramedian lobule. Cerebellar nuclear sections studied with the light microscope demonstrated a close relation of varicosities from BDA-labeled Purkinje cell axons with both gold lectin- and CTb-labeled neurons. Branches of individual axons could be traced to both retrogradely labeled cell populations. At the ultrastructural level, synapses of labeled Purkinje cell terminals with profiles of WGA-HRP-labeled projection neurons predominated over contacts with gold lectin-containing neurons. Nine out of 367 investigated BDA-labeled terminals were observed to be presynaptic to a WGA-HRP-labeled profile as well as to a gold lectin-labeled profile. This indicates that nuclear cells that project to the inferior olive as well as those that project to premotor centers are under the influence of the same Purkinje cells. Such an arrangement would suggest an in-phase cortical modulation of the activation patterns of the inhibitory cells that project to the inferior olive and excitatory cells that project to premotor nuclei, which could explain why olivary neurons, especially those of the rostral part of the dorsal accessory olive, appear to be unresponsive to stimuli generated during active movement. J. Comp. Neurol. 392:164–178, 1998. © 1998 Wiley-Liss, Inc.

The vestibulocerebellum is involved in the control of compensatory eye movements. To investigate its role in learning and timing of motor behavior, we investigated compensatory eye movements in mice with the use of search coils. Wild-type mice showed the ability to increase the gain of their vestibulo-ocular reflex by visuovestibular training. This adaptation did not occur in lurcher mice, a natural mouse mutant that completely lacks Purkinje cells. During the optokinetic reflex the phase of the eye movements of lurcher mice in reference to the stimulus lagged behind that of wild-type littermates, whereas during the vestibulo-ocular reflex it led that of the wild-type mice. During combined optokinetic and vestibular stimulation, the phase of the lurcher mice lagged behind that of the wild-type mice at the low stimulus frequencies, whereas it led the phase of the wild-type mice at the high frequencies. In addition, the optokinetic response of the lurcher mice showed a significantly longer latency during constant-velocity step stimulation than that of the wild-type mice. We conclude that Purkinje cells are necessary for both learning and timing of compensatory eye movements in mice. The present description of gain adaptation and phase dynamics provides the basis for studies in which the molecular mechanisms of cerebellar control of compensatory eye movements are investigated with the use of genetically manipulated mice.

The inferior olive provides climbing fibres to Purkinje cells in the cerebellar cortex and gives off axon collaterals to the cerebellar nuclei. The cerebellar nuclei contain GABAergic neurons that provide an inhibitory projection to the inferior olive and excitatory neurons that influence behaviour through various other premotor nuclei in the brainstem and diencephalon. Whether the olivary axon collaterals innervate the GABAergic neurons in the cerebellar nuclei is unknown. In the present study we investigated this projection in mice at the ultrastructural level using post-embedding GABA immunocytochemistry and anterograde and retrograde tracing of biotinylated dextrane amine and gold-lectin. It is demonstrated that the olivary axon collaterals do not only innervate non-GABAergic neurons in the cerebellar nuclei, but also GABAergic nucleo-olivary cells, thus establishing a direct feedback loop to the inferior olive.

The distribution of phospholipids across the two leaflets of the plasma membrane is important for many cellular processes including phagocytosis and hemostasis. In the present study we investigated the in vivo plasma membrane distribution of the aminophospholipid phosphatidylserine in mouse embryos with a novel technique employing Annexin V, a Ca2+ dependent phosphatidylserine binding protein, conjugated to fluorescein isothiocyanate and biotin. Annexin V directly applied to cryostat sections labeled the plasma membrane of all cells at the interface. In contrast, Annexin V injected intracardially into viable mouse embryos labeled almost exclusively apoptotic cells. These apoptotic cells were visible in all tissues and derived from all germ layers. Our experiments demonstrate that phosphatidylserine is asymmetrically distributed between the two leaflets of the plasma membrane in virtually all cell types in vivo and that this asymmetry is lost early during apoptosis.

De Zeeuw, C. I., S.K.E. Koekkoek, D.R.W. Wylie, and J. I. Simpson. Association between dendritic lamellar bodies and complex spike synchrony in the olivocerebellar system. J. Neurophysiol. 77: 1747–1758, 1997. Dendritic lamellar bodies have been reported to be associated with dendrodendritic gap junctions. In the present study we investigated this association at both the morphological and electrophysiological level in the olivocerebellar system. Because cerebellar GABAergic terminals are apposed to olivary dendrites coupled by gap junctions, and because lesions of cerebellar nuclei influence the coupling between neurons in the inferior olive, we postulated that if lamellar bodies and gap junctions are related, then the densities of both structures will change together when the cerebellar input is removed. Lesions of the cerebellar nuclei in rats and rabbits resulted in a reduction of the density of lamellar bodies, the number of lamellae per lamellar body, and the density of gap junctions in the inferior olive, whereas the number of olivary neurons was not significantly reduced. The association between lamellar bodies and electrotonic coupling was evaluated electrophysiologically in alert rabbits by comparing the occurrence of complex spike synchrony in different Purkinje cell zones of the flocculus that receive their climbing fibers from olivary subnuclei with different densities of lamellar bodies. The complex spike synchrony of Purkinje cell pairs, that receive their climbing fibers from an olivary subnucleus with a high density of lamellar bodies, was significantly higher than that of Purkinje cells, that receive their climbing fibers from a subnucleus with a low density of lamellar bodies. To investigate whether the complex spike synchrony is related to a possible synchrony between simple spikes, we recorded simultaneously the complex spike and simple spike responses of Purkinje cell pairs during natural visual stimulation. Synchronous simple spike responses did occur, and this synchrony tended to increase as the synchrony between the complex spikes increased. This relation raises the possibility that synchronously activated climbing fibers evoke their effects in part via the simple spike response of Purkinje cells. The present results indicate that dendritic lamellar bodies and dendrodendritic gap junctions can be downregulated concomitantly, and that the density of lamellar bodies in different olivary subdivisions is correlated with the degree of synchrony of their climbing fiber activity. Therefore these data support the hypothesis that dendritic lamellar bodies can be associated with dendrodendritic gap junctions. Considering that the density of dedritic lamellar bodies in the inferior olive is higher than in any other area of the brain, this conclusion implies that electrotonic coupling is important for the function of the olivocerebellar system.

The cerebellum is crucial for different types of motor learning. Established theories of cerebellar learning posit that the cerebellum learns by adjusting the weights of Parallel Fiber (PF) to Purkinje cells (PC) synapses, thanks to teaching signals provided by Climbing Fiber inputs. While these theories are consistent with a large body of experimental data, in particular on synaptic plasticity in PF to PC synapses, they cannot easily explain a growing body of experimental work, which seems to indicate a significant role of other sites of plasticity. Recent advances in the development of a large number in transgenic animals, as well as behavioral and electrophysiogical comparative studies between these animals and wild-type animals, have opened an unprecedented window into the mechanisms underlying learning in this structure. In particular, it has been shown that specific knock-outs are impaired selectively on difficult variants of the vestibulo-ocular reflex (VOR) adaptation task, one of the most studied cerebellar-dependent motor learning tasks. These impairments can occur even though the classical plasticity mechanisms are left untouched. These data pose significant new challenges for established models of cerebellar learning. To better understand the mechanisms of learning in the cerebellum, we built a model that can reproduce the available data on VOR adaptation, in both wild-type and transgenic animals. The model includes some of the main cell types involved in this task: granule cells (GCs), the input layer of cerebellar cortex, that receives vestibular information from the mossy fibers (MFs); Purkinje cells (PCs), as well as molecular layer interneurons (INs); and two cell populations in the medial vestibular nuclei (MVN), one excitatory and one inhibitory, that together control eye movement. The model also includes two sites of learning: the classical GC to PF plasticity site, as well as plasticity in the MF to MVN synapses. We provide a mechanistic understanding on how the system learns VOR adaptation in normal conditions, as well as how the system is impaired by specific knock-outs, which selectively suppress inhibition onto PCs, or increase the excitability of GCs. Finally, we show that our model is consistent with behavioral, as well as in vivo electrophysiological recordings.

It is well established that the olivocerebellar system is involved in the complex tasks of motor performance and motor learning, but the exact mechanism underlying these processes remains to be elucidated. The olivocerebellar system is equipped with electrical synapses at several locations. The electrical synapses provide the system with a tool that allows the formation or deformation of ensembles of neurons. These cooperating ensembles of neurons may be the key mechanism to encode large sets of complex motor instructions and/or they may coordinate temporal aspects of motor signals. This chapter describes all the locations of connexin36-mediated electrical synapses in the olivocerebellar system, their structural and electrophysiological signature, their functional and computational relevance, and their putative role(s) in the generation and optimization of motor behaviors.

The persistence of many contrasting notions of climbing fiber function after years of investigation testifies that the issue of climbing fiber contributions to cerebellar transactions is still unresolved. The proposed capabilities of the climbing fibers cover an impressive spectrum. For many researchers, the climbing fibers signal errors in motor performance, either in the conventional manner of frequency modulation or as a single announcement of an “unexpected event”. More controversial is the effect of these signals on the simple spike modulation of Purkinje cells. In some hands, they lead to a long-term depression of the strength of parallel fiber synapses, while, in other hands, they lead to a short-lasting enhancement of the responsiveness of Purkinje cells to mossy fiber inputs or contribute to the often-seen reciprocal relation between complex and simple spike modulation. For still other investigators, the climbing fibers serve internal timing functions through their capacity for synchronous and rhythmic firing. The above viewpoints are presented in the spirit of trying to reach some consensus about climbing fiber function. Each point of view is introduced by summarizing first the key observations made by the respective proponents; then the issues of short-lasting enhancement, reciprocity between complex and simple spikes, and synchrony and rhythmicity are addressed in the context of the visual climbing fiber system of the vestibulocerebellum.

The DBA/2J inbred strain of mice has been used extensively in hearing research as it suffers from early-onset, progressive hearing loss. Initially, it mostly affects high frequencies, but already at 2-3 months hearing loss becomes broad. In search for hearing loss genes other than Cadherin 23 (otocadherin) and fascin-2, which make a large contribution to the high-frequency deficits, we used a large set of the genetic reference population of BXD recombinant inbred strains. For frequencies 4, 8, 16 and 32 kHz, auditory brainstem response hearing thresholds were longitudinally determined from 2-3 up to 12 weeks of age. Apart from a significant, broad quantitative trait locus (QTL) for high-frequency hearing loss on chromosome 11 containing the fascin-2 gene, we found a novel, small QTL for low-frequency hearing loss on chromosome 18, from hereon called ahl9. Real-time quantitative polymerase chain reaction of organs of Corti, isolated from a subset of strains, showed that a limited number of genes at the QTL were expressed in the organ of Corti. Of those genes, several showed significant expression differences based on the parental line contributing to the allele. Our results may aid in the future identification of genes involved in low-frequency, early-onset hearing loss.

The projections of Purkinje cells from zones in the ventral nodulus of pigmented rabbits were studied with the use of extracellularly injected biocytin as an anterograde tracer. The zones were physiologically identified according to the complex spike modulation of Purkinje cells in response to optokinetic stimulation. Purkinje cells in the most medial zone do not respond to optokinetic stimulation; they project to the fastigial nucleus, the perifastigial white matter, the periinterposed white matter, and the medial vestibular nucleus. In the adjacent zone, Purkinje cells respond best to optokinetic stimulation about the vertical axis; they project to the periinterposed white matter and the medial vestibular nucleus. Purkinje cells in the next zone respond best to optokinetic stimulation about an axis approximately perpendicular to the ipsilateral anterior canal; they project to the periinterposed white matter, dorsal group y, the superior vestibular nucleus, and the medial vestibular nucleus. In the most lateral zone, Purkinje cells respond best to optokinetic stimulation about the vertical axis; they project to the periinterposed white matter, dorsal group y, and the medial vestibular nucleus. The majority of axons gave off collaterals and innervated more than one nucleus. Often, three or four different areas received terminals from a single Purkinje cell axon. The zonal projection pattern of the ventral nodulus is compared to that of the flocculus, which, with respect to the visual climbing fiber afferents, has similar zones.

Fragile X syndrome (FXS) is a single gene disorder that is the most common heritable cause of intellectual disability and the most frequent monogenic cause of autism spectrum disorders (ASD). FXS is caused by trinucleotide repeats in the promoter region of the fragile X mental retardation gene (fmr1). This leads to the downregulation of the fragile X mental retardation protein (FMRP), which regulates translation of a wide range of proteins. The extent of expression level alterations of synaptic proteins affected by FMRP loss and their consequences on synaptic dynamics in FXS has not been fully investigated. Here, we have investigated the molecular mechanisms underlying FXS by identifying the molecular signature and elucidating the function of FXS-associated proteins. The first part of the study consists of an exploratory molecular profiling study on brain tissue samples of a fmr1 KO mouse model using shotgun label-free liquid chromatography mass spectrometry (LC-MS). These studies indicate that most of the changed proteins are involved in synaptic functions. Thus, as a next step we investigated synaptosomes from the cerebellum and hippocampus using LC-MS and label-based selected reaction monitoring (SRM). Key findings relate to altered levels of proteins involved in GABA-signalling in the cerebellum. To explore these findings further, FM1-43 dye was used in cultured hippocampal neurons and cerebellar Purkinje cells to track vesicle recycling and unloading profiles. Furthermore, ultrastructural analysis of synaptic vesicles of the GABA-ergic Purkinje cells in the cerebellum was performed to investigate potential pre-synaptic effects. Taken together, these studies provide novel insights into the molecular changes associated with FXS.

1. We investigated saccadic eye movements elicited by microstimulation in the vermis of the rabbit. Scleral search coils were implanted under the conjunctiva of both eyes and a recording chamber was placed over the cerebellar vermis. 2. Conjugate saccadic eye movements were evoked in lobules VIa, b and c and VII of the vermis by currents ranging from 4 to 60 microA. All movements were horizontal with no apparent vertical component. 3. The cortex on both sides of the vermal mid-line could be divided in two zones, dependent on the direction of elicited saccades. In the medial zone saccades were directed ipsilaterally, in the lateral zone contralaterally. 4. We conclude that the topography of saccadic eye movements in the rabbit cerebellar vermis is, unlike in monkey and cat, organized in parasagittal zones.

The direct projection from the cerebellar nuclei to the inferior olive is GABAergic. In the present study, we investigated the projection from the cerebellar nuclei to the mesodiencephalic junction which is known to provide an excitatory projection to the inferior olive. The mesodiencephalic junction was studied in cat following anterograde transport of wheatgerm agglutinated horseradish peroxidase from the cerebellar nuclei in combination with: (1) retrograde transport of gold-lectin conjugate from the inferior olive; and (2) postembedding GABA-immunocytochemistry. Light microscopic analysis revealed that overlap of the anterograde and retrograde labeling was especially prominent in the nucleus of Darkschewitsch. Electron microscopic examination of this area showed: (1) that many cerebellar terminals made synaptic contacts with neurons that project to the inferior olive; (2) that virtually all cerebellar terminals were non-GABAergic and displayed an excitatory morphology; and (3) olivary projecting neurons were non-GABAergic. It is concluded that the indirect cerebellar projection to the inferior olive via the nucleus of Darkschewitsch is disynaptic and excitatory.

The dorsal cap and ventrolateral outgrowth of the inferior olive are involved in the control of eye movements. The caudal dorsal cap is predominantly involved in the horizontal optokinetic reflex; it receives most of its GABAergic input from the nucleus prepositus hypoglossi. In the present study, we determined the source of a major inhibitory input to the rostral dorsal cap and the ventrolateral outgrowth, which are the olivary subnuclei mainly involved in the "vertical" optokinetic reflexes. We studied these subnuclei in the rabbit with the use of retrograde tracing of horseradish peroxidase and anterograde tracing of wheat germ agglutinin-coupled horseradish peroxidase combined with postembedding immunocytochemistry. The ventral dentate nucleus of the cerebellum and dorsal group y project contralaterally to the rostral dorsal cap and ventrolateral outgrowth; this projection is entirely GABAergic. The terminals of this input form predominantly symmetric synapses with extraglomerular and intraglomerular dendrites; the remaining terminals are axosomatic. In addition, the dorsal cap and ventrolateral outgrowth contain significantly more crest synapses than any other olivary subnucleus. The terminals that form these crest synapses are derived from dorsal group y and/or the ventral dentate nucleus. None of the terminals in the dorsal cap or ventrolateral outgrowth was glycinergic.

The dorsal cap of the inferior olive is involved in the control of eye movements and is excited by inputs from the midbrain. In the present study we attempted to determine the inhibitory input to this nucleus in rat and rabbit. The projection from the nucleus prepositus hypoglossi to the dorsal cap was studied in the light microscope by anterograde tracing of Phaseolus vulgaris-leucoagglutinin and lesion-induced depletion of glutamic acid decarboxylase immunoreactivity, and in the electron microscope by anterograde tracing of wheat germ agglutinin-coupled horseradish peroxidase combined with GABA immunocytochemistry. We show that the nucleus prepositus hypoglossi projects bilaterally to the dorsal cap, contralaterally to the ventrolateral outgrowth, and ipsilaterally to the medial accessory olive. After lesioning of the nucleus prepositus hypoglossi, the caudal dorsal cap was depleted of most of its glutamic acid decarboxylase-immunoreactive terminals while the rostral dorsal cap and the ventrolateral outgrowth were depleted of a minor part. Ultrastructural analysis indicates that the majority, but not all, of the terminals from the nucleus prepositus hypoglossi in the dorsal cap are GABA-positive. These GABA-positive and GABA-negative terminals form predominantly symmetric and asymmetric synapses; most of them synapse on dendrites outside and inside glomeruli, frequently in association with dendrodendritic gap junctions, while a small minority are axosomatic. None of the terminals from the nucleus prepositus hypoglossi was found to form a crest synapse, although synapses of this kind were predominantly formed by GABAergic terminals. This study shows that the dorsal cap receives a major inhibitory input from the nucleus prepositus hypoglossi, the terminals of which are located at strategic positions on the olivary neurons.

Traditionally studies aimed at elucidating the molecular mechanisms underlying cerebellar motor learning have been focused on plasticity at the parallel fiber to Purkinje cell synapse. In recent years, however, the concept is emerging that formation and storage of memories are both distributed over multiple types of synapses at different sites. Here, we examined the potential role of potentiation at the mossy fiber to granule cell synapse, which occurs upstream to plasticity in Purkinje cells. We show that null-mutants of N-methyl D-aspartate-NR2A receptors (NMDA-NR2A(-/-) mice) have impaired induction of postsynaptic long-term potentiation (LTP) at the mossy fiber terminals and a reduced ability to raise the granule cell synaptic excitation, while the basic excitatory output of the mossy fibers is unaffected. In addition, we demonstrate that these NR2A(-/-) mutants as well as mutants in which the C terminal in the NR2A subunit is selectively truncated (NR2A(Delta C/Delta C) mice) have deficits in phase reversal adaptation of their vestibulo-ocular reflex (VOR), while their basic eye movement performance is similar to that of wild type littermates. These results indicate that NMDA-NR2A mediated potentiation at the mossy fiber to granule cell synapse is not required for basic motor performance, and they raise the possibility that it may contribute to some forms of vestibulo-cerebellar memory formation. (C) 2011 IBRO. Published by Elsevier Ltd. All rights reserved.

Grip force modulation has a rich history of research, but the results remain to be integrated as a neurocomputational model and applied in a robotic system. Adaptive grip force control as exhibited by humans would enable robots to handle objects with sufficient yet minimal force, thus minimizing the risk of crushing objects or inadvertently dropping them. We investigated the feasibility of grip force control by means of a biological neural approach to ascertain the possibilities for future application in robotics. As the cerebellum appears crucial for adequate grip force control, we tested a computational model of the olivo-cerebellar system. This model takes into account that the processing of sensory signals introduces a 100 ms delay, and because of this delay, the system needs to learn anticipatory rather than feedback control. For training, we considered three scenarios for feedback information: (1) grip force error estimation, (2) sensory input on deformation of the fingertips, and (3) as a control, noise. The system was trained on a data set consisting of force and acceleration recordings from human test subjects. Our results show that the cerebellar model is capable of learning and performing anticipatory grip force control closely resembling that of human test subjects despite the delay. The system performs best if the delayed feedback signal carries an error estimation, but it can also perform well when sensory data are used instead. Thus, these tests indicate that a cerebellar neural network can indeed serve well in anticipatory grip force control not only in a biological but also in an artificial system. (C) 2009 IBRO. Published by Elsevier Ltd. All rights reserved.

The fragile X syndrome (FRAXA) is the most widespread heritable form of mental retardation caused by the lack of expression of the fragile X mental retardation protein (FMRP). This lack has been related to deficits in cerebellum-mediated acquisition of conditioned eyelid responses in individuals with FRAXA. In the present behavioral study, long-term effects of deficiency of FMRP were investigated by examining the acquisition, savings and extinction of delay eyeblink conditioning in male individuals with FRAXA. In the acquisition experiment, subjects with FRAXA displayed a significantly poor performance compared with controls. In the savings experiment performed at least 6 months later, subjects with FRAXA and controls showed similar levels of savings of conditioned responses. Subsequently, extinction was faster in subjects with FRAXA than in controls. These findings confirm that absence of the FMRP affects cerebellar motor learning. The normal performance in the savings experiment and aberrant performance in the acquisition and extinction experiments of individuals with FRAXA suggest that different mechanisms underlie acquisition, savings and extinction of cerebellar motor learning.