J. van der Burg

Topography of cerebellar nuclear projections to the brain stem in the rat.

The organization of the cerebellum is characterized by a number of parallel and parasagittally ordered olivocorticonuclear modules; as such, the cerebellar nuclei basically function as output system of these modules. The present study provides a comprehensive and detailed description of the organization of the connections from the cerebellar nuclei to the brain stem in the rat. Thirteen small injections with the anterograde tracer Phaseolus vulgaris leucoagglutinin or biotinylated dextran amine which were centered on various aspects of the cerebellar nuclear complex are described and are illustrated with serial plots detailing the distribution of labeled varicosities throughout the brain stem. In every case at least 1,000 an up to 36,000 varicosities were plotted. All injections resulted in some or heavy labeling concentrated within specific regions of the contralateral inferior olivary complex and, usually, in some labeling of the contralateral ventrolateral thalamus. However, apart from these two areas it is shown that the cerebellar projections are generally very widespread and may be found throughout the entire brain stem. Below, only a survey of main projection areas will be given. Terminal arborizations originating from the rostral part of the medial cerebellar nucleus are mostly found in the caudal half of the brain stem with emphasis on the vestibular nuclear complex, whereas its caudal part rather connects to midbrain areas. Terminals that originate from the dorsolateral protuberance of the medial cerebellar nucleus are distributed more evenly throughout the brain stem and are mostly confined to reticular areas. The interstitial cell groups, interspersed between the medial and both interposed cerebellar nuclei, provide major projections to the ipsilateral vestibular nuclear complex and contralateral mesodiencephalic regions. However, reticular areas are also targeted over a large rostrocaudal range. The medial part of the posterior interposed nucleus sends most projections to the caudomedial red nucleus, prerubral regions and parvicellular reticular formation, all contralateral to the injection site. Projections that originate from more laterally placed injections are directed, apart from the inferior olivary complex, to the rostral half of the contralateral brain stem, where most labeled varicosities are found in the superior colliculus and zona incerta. The anterior interposed nucleus specifically targets the inferior olive, the red nucleus, the pontine reticulotegmental nucleus, the prectectum and the ventrolateral thalamic nucleus. More laterally placed injections also project to the ipsilateral parvicellular reticular formation and deep layers of the spinal trigeminal complex. The latter areas are more specifically targeted by the dorsolateral hump. In addition, its projections are found in the red nucleus and pretectum but do not seem to reach the ventrolateral thalamus. Projections from the lateral cerebellar nucleus are all characterized by a widespread distribution of terminals. Especially, the caudal aspect of the nucleus sends, apart from projections to the deep mesencephalic nucleus, red nucleus, periaquaductal gray, pretectum, prerubral area, and several thalamic regions, prominent projections to the caudal brain stem which terminate in the inferior olive and gigantocellular reticular formation. Projections from the ventral, parvicellular part of the nucleus are mostly, but not exclusively, directed to the rostral half of the brain stem and mainly terminate in the pararubral area, accessory oculomotor nuclei, pretectal areas, zona incerta, and in the parafascicular and ventrolateral thalamic nuclei. We conclude that the impact of the cerebellar nuclei on the brain stem is widespread; projections from different regions of the same cerebellar nucleus may show important differences in distribution of labeled terminals. On the other hand, injections placed in different cerebellar nuclei may result in a simila

Short-latency peripheral inputs to thalamic neurones projecting to the motor cortex in the monkey

SummaryOne hundred seventy-five neurones in the n.ventroposterior lateralis (VPL) and n.ventralis lateralis (VL) in the thalamus of anaesthetised monkeys have been tested antidromically for projection to the cortex and for somatosensory input from the contralateral arm.Using bipolar stimulation of the cortical surface, 113 thalamic neurones were successfully identified as antidromically driven from the hand area of the postcentral gyrus (48 neurones) or from the hand area of the precentral gyrus (65 neurones). All but one of these 113 neurones could only be antidromically discharged from the postcentral cortex or from the precentral cortex, and not from both. Most had antidromic latencies between 0.5 and 1.5 ms.Twenty-five/sixty-five precentrally projecting neurones and 45/48 postcentrally projecting neurones were activated by stimulation of the contralateral median or radial nerves. Both groups responded at short latency (4–8 ms) and many were activated by low-threshold shocks (0.8–1.3 T) and had restricted receptive fields on the hand. Precentrally projecting neurones responded most powerfully to joint movement or deep pressure, and some of these neurones were also responsive to cutaneous stimuli.Precentrally projecting neurones with peripheral inputs were all found in the oral subdivision of the VPL (the VPLo). The properties of these neurones suggest that they may be partly responsible for rapid somatosensory input to the motor cortex.

Purkinje cells in awake behaving animals operate at the upstate membrane potential

To the Editor: Over the last decades, cellular bistability or multistable states of the membrane potential have been demonstrated both in vitro and in vivo for different types of neurons throughout the brain, and various functions have been proposed for this phenomenon1,2. Recently, Loewenstein et al.3 proposed that bistability in Purkinje cells has a key role in the short-term processing and storage of sensorimotor information in the cerebellar cortex and that complex spikes may act as a toggle switch to control these processes. However, all intracellular recordings of bistability to date have been obtained either in slices or in anesthetized animals4. Because anesthetics can directly or indirectly affect the membrane potential5, it remains to be seen whether the proposed functional roles of bistability are valid in normal behaving animals under physiological conditions. To confirm the occurrence of bistability in Purkinje cells, we performed whole-cell patch recordings in vivo in mice under isoflurane or ketamine/xylazine anesthesia.

Single Purkinje Cell Can Innervate Multiple Classes of Projection Neurons in the Cerebellar Nuclei of the Rat: A Light Microscopic and Ultrastructural Triple-Tracer Study in the Rat

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.

The Function of Long Propriospinal Pathways in the Co-Ordination of Quadrupedal Stepping in the Cat

The functional organization and descending control of long propriospinal pathways, revealed in neurophysiological investigations in high spinal cats, were compared with the patterns of stepping and of quadrupedal co-ordination in normal and decerebrate cats. Emphasis was placed on the ascending propriospinal pathways and the coupling of hind-limb and forelimb movements.

A retrograde double-labeling technique for light microscopy. A combination of axonal transport of cholera toxin B-subunit and a gold-lectin conjugate.

A light microscopical, non-fluorescent, retrograde double-labeling technique is described. Cholera toxin B-subunit (CTb) and a conjugate of wheatgerm agglutinin and bovine serum albumin coupled to 10 nm gold particles (gold-lectin) are both excellent retrograde tracers and, when visualized by means of immunohistochemistry and silver intensification, respectively, may be readily identified within the same cell. This light microscopical retrograde double-labeling technique is illustrated in rat with experiments designed to investigate the collateralisation (1) of vestibular neurons to the spinal cord and oculomotor complex, (2) of spinal neurons to the left and right lateral reticular nucleus, and (3) of inferior olivary neurons to the uvula of the cerebellum. Advantages over fluorescent double-labeling experiments are found in the fact that the diaminobenzidine reaction product as well as the silver/gold deposits do not fade and can be examined in counterstained sections. Moreover, the injection sites can be kept quite small and may be guided by electrophysiological recording through the injection pipette.

Intrathecal injection of GDNF and BDNF induces immediate early gene expression in rat spinal dorsal horn

Glial cell line-derived neurotrophic factor (GDNF) and brain-derived neurotrophic factor (BDNF) are potent trophic factors for dorsal root ganglion cells. In addition, these factors are produced in subsets of dorsal root ganglion cells and transported anterogradely to their terminals in the superficial dorsal horn of the spinal cord, where they constitute the only source of GDNF and BDNF. We investigated the effect of 10 mug GDNF and BDNF injected by lumbar puncture on the expression of the immediate early gene (IEG) products c-Fos, c-Jun, and Krox-24 in the adult rat dorsal horn. In the dorsal horn of S1 spinal segments, GDNF and BDNF induced a strong increase in IEG expression, which was most pronounced in laminae I and II (2.9- to 4.5-fold). More distal from the injection site, in the dorsal horn of L1/L2 spinal segments, the increase in IEG expression was less pronounced, suggesting a concentration-dependent effect. In order to explain the effects of intrathecally injected GDNF, we investigated whether lumbo-sacral dorsal horn neurons expressed RET protein, the signal-transducing element of the receptor complex for GDNF. It was found that several of these neurons contained RET immunoreactivity and that some of the RET-labeled neurons had the appearance of nociceptive-specific cells, confirming their presumed role in pain transmission. Additionally, using double-labeling immunofluorescence combined with confocal microscopy, it was found that after intrathecal GDNF injection 35% of c-Fos-labeled cells were also labeled for RET. These results demonstrate that intrathecally administered GDNF and BDNF induce IEG expression in dorsal horn neurons in the adult rat, supposedly by way of their cognate receptors, which are present on these neurons. We further suggest that the endogenous release of GDNF and BDNF, triggered by nociceptive stimuli, is involved in the induction of changes in spinal nociceptive transmission as in various pain states.

Topography of saccadic eye movements evoked by microstimulation in rabbit cerebellar vermis.

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.

Modulation of cutaneous reflexes in hindlimb muscles during locomotion in the freely walking rat: A model for studying cerebellar involvement in the adaptive control of reflexes during rhythmic movements

This study aims to demonstrate stepphase-dependent modulation in the gain of cutaneously triggered reflexes in the freely locomoting rat. Electromyographic recordings of biceps femoris (mainly involved in knee flexion) and gastrocnemius (mainly involved in ankle extension) muscles were continuously monitored during locomotion and cutaneous reflexes were induced by subcutaneously placed stimulation electrodes in the lateral malleolal region. The results show that the reflex responses in both muscles during locomotion were generally reduced compared to reflexes induces in rest. For the biceps femoris reduction of reflex gain was highest during the stance phase whereas for the gastrocnemius the period of highest depression was found during the swing phase. We conclude that stepphase-dependent modulation of peripheral reflexes can be measured in freely locomoting rats and generally concur with previous studies in cat and man that this type of modulation may be functionally important for maintaining and adjusting gait. Moreover, although the mechanism of inducing and maintaining this modulation is not fully known, it is now open to experimental investigation in rodents.