Inferior Olivary Axons Research Articles

Pax3 induces target-specific reinnervation through axon collateral expression of PSA-NCAM

Damaged or dysfunctional neural circuits can be replaced after a lesion by axon sprouting and collateral growth from undamaged neurons. Unfortunately, these new connections are often disorganized and rarely produce clinical improvement. Here we investigate how to promote post-lesion axonal collateral growth, while retaining correct cellular targeting. In the mouse olivocerebellar path, brain-derived neurotrophic factor (BDNF) induces correctly-targeted post-lesion cerebellar reinnervation by remaining intact inferior olivary axons (climbing fibers). In this study we identified cellular processes through which BDNF induces this repair. BDNF injection into the denervated cerebellum upregulates the transcription factor Pax3 in inferior olivary neurons and induces rapid climbing fiber sprouting. Pax3 in turn increases polysialic acid-neural cell adhesion molecule (PSA-NCAM) in the sprouting climbing fiber path, facilitating collateral outgrowth and pathfinding to reinnervate the correct targets, cerebellar Purkinje cells. BDNF-induced reinnervation can be reproduced by olivary Pax3 overexpression, and abolished by olivary Pax3 knockdown, suggesting that Pax3 promotes axon growth and guidance through upregulating PSA-NCAM, probably on the axon’s growth cone. These data indicate that restricting growth-promotion to potential reinnervating afferent neurons, as opposed to stimulating the whole circuit or the injury site, allows axon growth and appropriate guidance, thus accurately rebuilding a neural circuit.

Electrical coupling regulated by GABAergic nucleo-olivary afferent fibres facilitates cerebellar sensory–motor adaptation

The inferior olivary (IO) nucleus makes up the signal gateway for several organs to the cerebellar cortex. Located within the sensory–motor-cerebellum pathway, the IO axons, i.e., climbing fibres (CFs), massively synapse onto the cerebellar Purkinje cells (PCs) regulating motor learning whilst the olivary nucleus receives negative feedback through the GABAergic nucleo-olivary​ (NO) pathway. The NO pathway regulates the electrical coupling (EC) amongst the olivary cells thus facilitating synchrony and timing. However, the involvement of this EC regulation on cerebellar adaptive behaviour is still under debate. In our study we have used a spiking cerebellar model to assess the role of the NO pathway in regulating vestibulo-ocular-reflex (VOR) adaptation. The model incorporates spike-based synaptic plasticity at multiple cerebellar sites and an electrically-coupled olivary system. The olivary system plays a central role in regulating the CF spike-firing patterns that drive the PCs, whose axons ultimately shape the cerebellar output. Our results suggest that a systematic GABAergic NO deactivation decreases the spatio-temporal complexity of the IO firing patterns thereby worsening the temporal resolution of the olivary system. Conversely, properly coded IO spatio-temporal firing patterns, thanks to NO modulation, finely shape the balance between long-term depression and potentiation, which optimises VOR adaptation. Significantly, the NO connectivity pattern constrained to the same micro-zone helps maintain the spatio-temporal complexity of the IO firing patterns through time. Moreover, the temporal alignment between the latencies found in the NO fibres and the sensory–motor pathway delay appears to be crucial for facilitating the VOR. When we consider all the above points we believe that these results predict that the NO pathway is instrumental in modulating the olivary coupling and relevant to VOR adaptation.

Uncoupling axon guidance and neuronal migration in Robo3-deficient inferior olivary neurons.

Inferior olivary (IO) neurons are born in the dorsal hindbrain and migrate tangentially toward the ventral midline. During their dorsoventral migration, IO neurons extend long leading processes that cross the midline, transform into axons, and project into the contralateral cerebellum. In absence of the axon guidance receptor Robo3, IO axons fail to cross the midline and project to the ipsilateral cerebellum. Remarkably, the IO cell bodies still reach the midline where they form a nucleus of abnormal cytoarchitecture. The mechanisms underlying the migration of Robo3-deficient IO neurons are unknown. Here, we used three-dimensional imaging and transgenic mice to label subsets of IO neurons and study their migratory behavior in Robo3 knockout. We show that IO migration is delayed in absence of Robo3. Strikingly, Robo3-deficient IO neurons progress toward the midline in a direction opposite to their axons. This occurs through a change of polarity and the generation of a second leading process at the rear of the cell. These results suggest that Robo3 receptor controls the establishment of neuronal polarity and the coupling of axonogenesis and cell body migration in IO neurons.

Synaptic excitation by climbing fibre collaterals in the cerebellar nuclei of juvenile and adult mice.

The inferior olive sends instructive motor signals to the cerebellum via the climbing fibre projection, which sends collaterals directly to large premotor neurons of the mouse cerebellar nuclei (CbN cells). Optogenetic activation of inferior olivary axons in vitro evokes EPSCs in CbN cells of several hundred pA to more than 1nA. The inputs are three-fold larger at younger ages, 12 to 14days old, than at 2months old, suggesting a strong functional role for this pathway earlier in development. The EPSCs are multipeaked, owing to burst firing in several olivary afferents that fire asynchronously. The convergence of climbing fibre collaterals onto CbN cells decreases from ∼40 to ∼8, which is consistent with the formation of closed-loop circuits in which each CbN neuron receives input from 4-7 collaterals from inferior olivary neurons as well as from all 30-50 Purkinje cells that are innervated by those olivary neurons. The inferior olive conveys instructive signals to the cerebellum that drive sensorimotor learning. Inferior olivary neurons transmit their signals via climbing fibres, which powerfully excite Purkinje cells, evoking complex spikes and depressing parallel fibre synapses. Additionally, however, these climbing fibres send collaterals to the cerebellar nuclei (CbN). In vivo and in vitro data suggest that climbing fibre collateral excitation is weak in adult mice, raising the question of whether the primary role of this pathway may be developmental. We therefore examined climbing fibre collateral input to large premotor CbN cells over development by virally expressing channelrhodopsin in the inferior olive. In acute cerebellar slices from postnatal day (P)12-14 mice, light-evoked EPSCs were large (>1nA at -70mV). The amplitude of these EPSCs decreased over development, reaching a plateau of ∼350pA at P20-60. Trains of EPSCs (5Hz) depressed strongly throughout development, whereas convergence estimates indicated that the total number of functional afferents decreased with age. EPSC waveforms consisted of multiple peaks, probably resulting from action potential bursts in single collaterals and variable times to spike threshold in converging afferents. Activating climbing fibre collaterals evoked well-timed increases in firing probability in CbN neurons, especially in younger mice. The initially strong input, followed by the decrement in synaptic strength coinciding with the pruning of climbing fibres in the cerebellar cortex, implicates the climbing fibre collateral pathway in early postnatal development. Additionally, the persistence of substantial synaptic input at least to P60 suggests that this pathway may function in cerebellar processing into adulthood.

Dendrites of cerebellar granule cells correctly recognize their target axons for synaptogenesis in vitro

Neural circuits are generated by precisely ordered synaptic connections among neurons, and this process is thought to rely on the ability of neurons to recognize specific partners. However, it is also known that neurons promiscuously form synapses with nonspecific partners, in particular when cultured in vitro, causing controversies about neural recognition mechanisms. Here we reexamined whether neurons can or cannot select particular partners in vitro. In the cerebellum, granule cell (GC) dendrites form synaptic connections specifically with mossy fibers, but not with climbing fibers. We cocultured GC neurons with pontine or inferior olivary axons, the major sources for mossy and climbing fibers, respectively, as well as with hippocampal axons as a control. The GC neurons formed synapses with pontine axons predominantly at the distal ends of their dendrites, reproducing the characteristic morphology of their synapses observed in vivo, whereas they failed to do so when combined with other axons. In the latter case, synaptic proteins could accumulate between axons and dendrites, but these synapses were randomly distributed throughout the contact sites, and also their synaptic vesicle recycling was anomalous. These observations suggest that GC dendrites can select their authentic partners for synaptogenesis even in vitro, forming the synapses with a GC-specific nature only with them.

BDNF increases homotypic olivocerebellar reinnervation and associated fine motor and cognitive skill

Recovery of complex neural function after injury to the adult CNS is limited by minimal spontaneous axonal regeneration and/or sprouting from remaining pathways. In contrast, the developing CNS displays spontaneous reorganization following lesion, in which uninjured axons can develop new projections to appropriate target neurons and provide partial recovery of complex behaviours. Similar pathways can be induced in the mature CNS, providing models to optimize post-injury recovery of complex neural functions. After unilateral transection of a developing olivocerebellar path (pedunculotomy), remaining inferior olivary axons topographically reinnervate the denervated hemicerebellum and compensate functional deficits. Brain-derived neurotrophic factor (BDNF) partly recreates such reinnervation in the mature cerebellum. However the function of this incomplete reinnervation and any unwanted behavioural effects of BDNF remain unknown. We measured olivocerebellar reinnervation and tested rotarod and navigation skills in Wistar rats treated with BDNF/vehicle and pedunculotomized on day 3 (Px3; with reinnervation) or 11 (Px11; without spontaneous reinnervation). BDNF treatment did not affect motor or spatial behaviour in normal (control) animals. Px11-BDNF animals equalled controls on the rotarod, outperforming Px11-vehicle animals. Moreover, Px3-BDNF and Px11-BDNF animals achieved spatial learning and memory tasks as well as controls, with Px11-BDNF animals showing better spatial orientation than Px11-vehicle counterparts. BDNF slightly increased olivocerebellar reinnervation in Px3 animals and induced sparse (22% Purkinje cells) yet widespread reinnervation in Px11 animals. As reinnervation correlated with spatial function, these data imply that after injury even a small amount of reinnervation that is homotypic to correct target neurons compensates deficits in appropriate complex motor and spatial skills. As there was no effect in control animals, BDNF effectively induces this axon collateralisation without interfering with normal neuronal circuits.

Signals from the cerebellum guide the pathfinding of inferior olivary axons

During development, inferior olivary axons cross the floor plate and project from the caudal to the rostral hindbrain, whence they grow into the cerebellar plate. We have investigated the axon guidance signals involved in the formation of this projection in vitro. When the cerebellar plate was grafted ectopically along the margin of the hindbrain in organotypic cultures, inferior olivary axons could pathfind to the ectopic cerebellum, establishing a topographically normal projection. Following rostrocaudal reversal of a region of tissue in the axon pathway between the inferior olive and the cerebellum, olivary axons still navigated towards the cerebellum. Moreover, olivary axons could cross a bridging tissue explant (spinal cord) to reach a cerebellar explant. In collagen gel cultures of inferior olive explants, olivary axon outgrowth increased significantly in the presence of cerebellar explants and axons deflected towards the cerebellar tissue. These results show that the cerebellum is a source of diffusible axon guidance signals for olivary axons. We also found that, in organotypic cultures, olivary axons which had crossed the floor plate showed an increased tendency to respond to cerebellar cues. Taken together, these results indicate that the cerebellum is the source of cues that are chemoattractant and growth-promoting for inferior olivary axons; prior exposure to the floor plate increases responsiveness to these cues.

Postnatal development and adult organisation of the olivocerebellar projection map in the hypogranular cerebellum of the rat.

The olivocerebellar system is characterised by a precise topographical organisation, in which distinct subsets of inferior olivary axons project to neurochemically heterogeneous Purkinje cell subpopulations, arranged into parasagittally oriented compartments in the cerebellar cortex. Adult climbing fibres and Purkinje cells are linked by a one-to-one relationship, which is established during postnatal development after a transitory phase of multiple climbing fibre innervation. The elimination of redundant climbing fibre synapses is thought to be regulated by granule cell-mediated activity-dependent processes. In order to assess whether this developmental remodelling is also important for the construction of the mature olivocerebellar projection map, we examined the hypogranular cerebella of rats treated by means of methylazoxymethanol acetate (MAM) during early postnatal life, in which multiple climbing fibre innervation persists in the adult. In these animals we investigated the distribution of calcitonin gene-related peptide (CGRP)-immunoreactive olivocerebellar axons and arbours during early postnatal development, and the correspondence between climbing fibre strips and zebrin II-defined Purkinje cell bands in the adult. Our results show that: (1) the pattern of CGRP-immunoreactive climbing fibres observed during the first three postnatal weeks is not disrupted after granule cell degeneration; and (2) the alignment between olivocerebellar axon subsets and zebrin II+/- Purkinje cell compartments is normally achieved in adult rats. In contrast, the climbing fibre-Purkinje cell relationship is abnormal, and single arbours innervate restricted dendritic regions of several neighbouring target neurons. These results indicate that the normal distribution of olivocerebellar axon subsets to distinct cerebellar cortical compartments can be established independently from granule cell-mediated remodelling processes. Thus, the postnatal climbing fibre plasticity, which is needed to achieve the normal climbing fibre-Purkinje cell relationship, appears to be confined within the framework of a projection map established during earlier developmental phases.

Reestablishment of the olivocerebellar projection map by compensatory transcommissural reinnervation following unilateral transection of the inferior cerebellar peduncle in the newborn rat.

It is unclear whether reparative processes in the injured mammalian brain are able to restore the topographic organisation of neuronal connections. To address this question, we have investigated the plasticity of the olivocerebellar system. This pathway has a precise topographic arrangement, in which subsets of inferior olivary neurons project to parasagittally oriented Purkinje cell compartments. Following unilateral transection of the inferior cerebellar peduncle in newborn rats, axons from the contralateral projection cross the cerebellar midline and reinnervate the deafferented hemicerebellum. By this experimental approach, we first analysed the behaviour of calcitonin gene-related peptide (CGRP)-immunoreactive climbing fibres. This marker is transiently expressed by a subset of developing inferior olivary axons, which terminate in the cerebellar cortex into several parasagittal strips. We show that transcommissural axons reestablish the original pattern of climbing fibre bands within a few days after lesion. Then, in adult animals injured at birth, we assessed whether the newly formed climbing fibre bands align with zebrin II+/- Purkinje cell compartments, as in normal conditions. The newly formed projection is organised in parasagittally oriented strips which mirror the distribution of their counterparts on the intact side and are precisely aligned to the heterogeneous Purkinje cell compartments. In addition, the patchy distribution of olivo-nuclear fibres suggests that specific reinnervation is also achieved in the deep nuclei. Thus, transcommissural olivocerebellar reinnervation is not random, but it is regulated by selective interactions between distinct subsets of olivocerebellar axons and target neurons aimed at reestablishing the correct projection map.

The embryonic cerebellum contains topographic cues that guide developing inferior olivary axons.

The formation of the olivocerebellar projection is supposed to be regulated by positional information shared between pre- and postsynaptic neurons. However, experimental evidence to support this hypothesis is missing. In the chick, caudal neurons in the inferior olive project to the anterior cerebellum and rostral ones to the posterior cerebellum. We here report in vitro experiments that strongly support the existence of anteroposterior polarity cues in the embryonic cerebellum. We developed an in vitro system that was easily accessible to experimental manipulations. Large hindbrain explants of E7.5-E8 chick embryos, containing the cerebellum and its attached brainstem, were plated and studied using axonal tracing methods. In these cultures, we have shown that the normal anteroposterior topography of the olivocerebellar projection was acquired, even when the cerebellar lamella was detached from the brainstem and placed again in its original position. We also found that, following various experimental rotations of the anteroposterior axis of the cerebellum, the rostromedian olivary neurons still project to the posterior vermis and the caudolateral neurons to the anterior vermis, that now have inverted locations. Thus, the rotation of the target region results in the rotation of the projection. In addition, we have shown that the formation of the projection map could be due to the inability of rostromedian inferior olivary axons to grow in the anterior cerebellum. All these experiments strongly indicate that olivocerebellar fibers recognize within their target region polarity cues that organize their anteroposterior topography, and we suggest that Purkinje cells might carry these cues.

BEN As a Presumptive Target Recognition Molecule during the Development of the Olivocerebellar System

It has been shown previously that in the chick embryo the cell adhesion molecule BEN/SC1/DM-GRASP is expressed by neurons in the inferior olive (IO) and by their terminal axonal arbors in the cerebellar cortex, the climbing fibers (Porquié et al., 1992b). Here, new information on the expression of BEN during the formation of the olivocerebellar projection adds the important notion that BEN is also expressed by the cerebellar targets of inferior olivary axons, Purkinje cells (PCs) and deep nuclear neurons. This expression is transient, starting at E7-E8 and vanishing shortly after hatching. More importantly, BEN expression is restricted to precise subsets of IO neurons and PCs. In the cerebellar cortex, BEN-immunoreactive (BEN-IR) structures are not found randomly but are distributed according to a reproducible pattern of parasagittal stripes. A maximum of four distinct sagittal stripes is found in each lobule, along the whole rostrocaudal extent of the cerebellum. Moreover, BEN-expressing stripes belong to two classes; one contains BEN-IR climbing fibers terminating on BEN-IR PCs and the other, more frequent class is solely composed of BEN-IR climbing fibers. Organotypic cultures of isolated cerebella have shown that the expression of BEN in the IO and in the cerebellum arise independently, probably because of an intrinsic developmental program. Thus, the cell adhesion molecule BEN meets all criteria for a recognition molecule involved in the formation of the olivocerebellar projection.

Branching of inferior olivary axons to terminate in different folia, lobules or lobes of the cerebellum

Branching of inferior olivary axons to terminate in different folia, lobules or lobes of the cerebellum

Relations among climbing fiber responses of nearby Purkinje Cells.

Relations among climbing fiber responses of nearby Purkinje Cells.C C Bell, and T KawasakiC C Bell, and T KawasakiPublished Online:01 Mar 1972https://doi.org/10.1152/jn.1972.35.2.155MoreSectionsPDF (2 MB)Download PDF ToolsExport citationAdd to favoritesGet permissionsTrack citations ShareShare onFacebookTwitterLinkedInWeChat Back to Top Next Download PDF FiguresReferencesRelatedInformation Cited ByCoding of stimulus strength via analog calcium signals in Purkinje cell dendrites of awake mice9 September 2014 | eLife, Vol. 3Computational model of an adaptive rhythm generator within the olivocerebellar system9 July 2009 | Network: Computation in Neural Systems, Vol. 16, No. 1Clustering behavior in a three-layer system mimicking olivo-cerebellar dynamicsNeural Networks, Vol. 17, No. 2Developmental changes in eyeblink conditioning and simple spike activity in the cerebellar cortex1 January 2003 | Developmental Psychobiology, Vol. 44, No. 1Dynamical Working Memory and Timed Responses: The Role of Reverberating Loops in the Olivo-Cerebellar SystemNeural Computation, Vol. 14, No. 11Generation and Propagation of Subthreshold Waves in a Network of Inferior Olivary NeuronsAnna Devor, and Yosef Yarom1 June 2002 | Journal of Neurophysiology, Vol. 87, No. 6Coherent Electrical Activity Between Vibrissa Sensory Areas of Cerebellum and Neocortex Is Enhanced During Free WhiskingSean M. O'Connor, Rune W. Berg, and David Kleinfeld1 April 2002 | Journal of Neurophysiology, Vol. 87, No. 4Chapter 19 The cerebellum as a neuronal prosthesis machineSerotonin suppresses subthreshold and suprathreshold oscillatory activity of rat inferior olivary neurones in vitro13 August 2004 | The Journal of Physiology, Vol. 524, No. 3Time window control: a model for cerebellar function based on synchronization, reverberation, and time slicingSynchronized Oscillations in the Inferior Olive Are Controlled by the Hyperpolarization-Activated Cation Current IhThierry Bal, and David A. McCormick1 June 1997 | Journal of Neurophysiology, Vol. 77, No. 6The effect of bicarbonate-free artificial cerebrospinal fluid on spontaneous oscillations of the membrane potential in inferior olivary neurons of the ratBrain Research, Vol. 639, No. 1Zonal organization of climbing fiber projections to the nodulus in the catBrain Research, Vol. 638, No. 1-2Effects of recurrent collateral inhibition on Purkinje cell activity in the immature rat cerebellar cortex - an in vivo electrophysiological studyBrain Research, Vol. 626, No. 1-2Development and properties of spontaneous oscillations of the membrane potential in inferior olivary neurons in the ratDevelopmental Brain Research, Vol. 65, No. 1Stimulus classification by ensembles of climbing fiber receptive fieldsTrends in Neurosciences, Vol. 14, No. 6Oscillatory behavior in inferior olive neurons: Mechanism, modulation, cell aggregatesBrain Research Bulletin, Vol. 17, No. 6Zonal organization of olivocerebellar projections to the uvula in rabbitsBrain Research, Vol. 359, No. 1-2Electrophysiological study of the corticonuclear projection in the cat cerebellumBrain Research, Vol. 327, No. 1-2Rhythmic activity of spinal interneurons in harmaline-treated catsJournal of the Neurological Sciences, Vol. 54, No. 3Zonal organization of the floccular Purkinje cells projecting to the vestibular nucleus in catsBrain Research, Vol. 232, No. 1Regression of functional synapses in the immature mammalian cerebellumTrends in Neurosciences, Vol. 5The olivocerebellar system. I. Delayed and slow inhibitory effects: An overlooked salient feature of cerebellar climbing fibersBrain Research, Vol. 187, No. 1Storing covariance with nonlinearly interacting neuronsJournal of Mathematical Biology, Vol. 4, No. 4Studies on field potentials and on single cells in the inferior olivary complex of the ratBrain Research, Vol. 101, No. 3Drug-induced rhythmical activity in the inferior olivary complex of the ratBrain Research, Vol. 101, No. 3Cerebellar responses to teleceptive stimuli in alert monkeysBrain Research, Vol. 83, No. 3A theory of memory that explains the function and structure of the cerebellumBrain Research, Vol. 70, No. 1Branching of inferior olivary axons to terminate in different folia, lobules or lobes of the cerebellumBrain Research, Vol. 54Rhythmic activity induced by harmaline in the olivo-cerebello-bulbar system of the catBrain Research, Vol. 53, No. 1Cerebellar afferent systems: A reviewProgress in Neurobiology, Vol. 2Climbing fibre Purkinje cell twins are foundClimbing fiber Purkinje cells twins: a search of extra-rare events in neural systems More from this issue > Volume 35Issue 2March 1972Pages 155-69 https://doi.org/10.1152/jn.1972.35.2.155PubMed4337637History Published online 1 March 1972 Published in print 1 March 1972 Metrics