Scanning electron microscopy of human cerebellar cortex

Summary Scanning electron microscopy and cryofracture technique were applied to study neuronal architecture and synaptic connections of the human cerebellum. Samples were processed according to the technique of Humphreys et al. (1975) with minor modifications. The granule cells exhibit unbranched filiform axons and coniform dendritic processes. The latter show typical claw-like endings making gearing type synaptic contacts with mossy fiber rosettes. The unattached mossy rosettes appear as solid club-like structures. Some fractographs show individual granule cells, Golgi neurons and glomerular islands. The climbing fibers and their Scheibel's collaterals were also characterized. In the Purkinje layer the surface fracture was produced at the level of the Bergmann glial cells, which are selectively removed, allowing us to visualize the rough surface of Purkinje cells and the supra- and infraganglionic plexuses of basket cell axons which appeared as entangled threads. In the molecular layer the three-dimensional configuration of the Purkinje secondary and tertiary dendritic branches was obtained. The filiform parallel fibers make cruciform synaptic contacts with the Purkinje dendritic spines. The appearance of stellate neuronal somata closely resembled that of the granule cells. The subpial terminals of Bergmann fibers appeared attached to the exterior of the folia forming the rough surfaced external glial limiting membrane.     

Formation of new synaptic contacts by Purkinje axon collaterals in the granular layer of deafferented cerebellar cortex of adult rat

In addition to (i) mossy terminals, (ii) Golgi axons, (iii) granule cell dendrites and (iv), occasionally, Golgi cell dendrites, a third axonal profile identified by morphological criteria as the collateral of Purkinje axons, has been found in 2% of all cerebellar glomeruli. These infrequent components of a few glomeruli, however, were never seen in normal cerebellar cortex to establish specialized synaptic contact with glomerular dendrites. Two to four weeks after surgical isolation of the cerebellar cortex, i.e. following the destruction of both efferent and afferent fibres, the number of glomeruli containing (hypertrophic) axonal branches of Purkinje cells has increased to 13% of all surveyed glomeruli. In addition, the Purkinje axon terminals in the mossy fibre-deprived glomeruli were observed to establish numerous Gray II-type synaptic contacts with surrounding granule cell dendrites. It is suggested that the development of heterologous synapses between hypertrophic, or even intact, Purkinje axon collaterals on the one hand and the mossy fibre-vacated granule cell dendrites on the other, is a compensatory, reactive process to the synaptic "desaturation" of granule neurons, which demonstrate a dormant potential of Purkinje cells to form new synaptic contacts in the adult cerebellum.     

Morphological study of organotypic cerebellar cultures

Organotypic cerebellar cultures from 8-days-old (P8) mouse pups were studied following 11 days of in vitro (I IDIV) culturing. The cerebellar cytoarchitectonic structure was maintained in most parasagittal cerebellar cortical slice cultures (also containing the deep cerebellar nuclei). The two main extrinsic excitatory inputs (the climbing and the mossy fibers) seem to be replaced by other axonal types: in the molecular layer mostly by parallel fibers (for climbing fibers) and in the granular layer by intrinsic mossy fiber collaterals of local excitatory interneurons, the unipolar brush cells. However, in a few organotypic cultures, which (although preserving the trilaminar cerebellar cortical structure) were "granuloprival" but also contained some of the deep cerebellar nuclei, the participation of extracortical axons from the deep cerebellar nuclei in the replacement of the missing afferents is suggested.     

Cerebellar globular cells receive monoaminergic excitation and monosynaptic inhibition from Purkinje cells

Inhibitory interneurons in the cerebellar granular layer are more heterogeneous than traditionally depicted. In contrast to Golgi cells, which are ubiquitously distributed in the granular layer, small fusiform Lugaro cells and globular cells are located underneath the Purkinje cell layer and small in number. Globular cells have not been characterized physiologically. Here, using cerebellar slices obtained from a strain of gene-manipulated mice expressing GFP specifically in GABAergic neurons, we morphologically identified globular cells, and compared their synaptic activity and monoaminergic influence of their electrical activity with those of small Golgi cells and small fusiform Lugaro cells. Globular cells were characterized by prominent IPSCs together with monosynaptic inputs from the axon collaterals of Purkinje cells, whereas small Golgi cells or small fusiform Lugaro cells displayed fewer and smaller spontaneous IPSCs. Globular cells were silent at rest and fired spike discharges in response to application of either serotonin (5-HT) or noradrenaline. The two monoamines also facilitated small Golgi cell firing, but only 5-HT elicited firing in small fusiform Lugaro cells. Furthermore, globular cells likely received excitatory monosynaptic inputs through mossy fibers. Because globular cells project their axons long in the transversal direction, the neuronal circuit that includes interplay between Purkinje cells and globular cells could regulate Purkinje cell activity in different microzones under the influence of monoamines and mossy fiber inputs, suggesting that globular cells likely play a unique modulatory role in cerebellar motor control.     

Physiological properties of afferents and synaptic reorganization in the rat cerebellum degranulated by postnatal x-irradiation

Elimination of most granule, basket, and stellate interneurons in the rat cerebellum was achieved by repeated doses of low level x-irradiation applied during the first two weeks of postnatal life. Electrical stimulation of the brain stem and peripheral limbs was employed to investigate the properties of afferent cerebellar pathways and the nature of the reorganized neuronal synaptic circuitry in the degranulated cerebellum of the adult. Direct contacts of mossy fibers on Purkinje cells were indicated by short latency, single spike responses: 1.9 msec from the lateral reticular nucleus of brain stem and 5.4 msec from ipsilateral forlimb. These were shorter than in normal rats by 0.9 and 2.1 msec, respectively. The topography of projections from peripheral stimulation was approximately normal. Mossy fiber responses followed stimulation at up to 20/sec, whereas climbing fiber pathways fatigued at 10/sec. The latency of climbing fiber input to peripheral limb stimulation in x-irradiated cerebellum was 23 ± 8 (SD) msec. In x-irradiated rats, the climbing fiber pathways evoked highly variable extracellular burst responses and intracellular EPSPs of different, discrete sizes. These variable responses suggest that multiple climbing fibers contact single Purkinje cells. We conclude that each type of afferent retains identifying characteristics of transmission. However, rules for synaptic specification appear to break down so that: (1) abnormal classes of neurons develop synaptic connections, i.e., mossy fibers to Purkinje cells; (2) incorrect numbers of neurons share postsynaptic targets, i.e., more than one climbing fiber to a Purkinje cell; and (3) inhibitory synaptic actions may be carried out in the absence of the major inhibitory interneurons, i.e., Purkinje cell collaterals may be effective in lieu of basket and stellate cells.     

Physiological properties of afferents and synaptic reorganization in the rat cerebellum degranulated by postnatal X-irradiation.

Elimination of most granule, basket, and stellate interneurons in the rat cerebellum was achieved by repeated doses of low level x-irradiation applied during the first two weeks of postnatal life. Electrical stimulation of the brain stem and peripheral limbs was employed to investigate the properties of afferent cerebellar pathways and the nature of the reorganized neuronal synaptic circuitry in the degranulated cerebellum of the adult. Direct contacts of mossy fibers on Purkinje cells were indicated by short latency, single spike responses: 1.9 msec from the lateral reticular nucleus of brain stem and 5.4 msec from ipsilpateral forelimb. These were shorter than in normal rats by 0.9 and 2.1 msec, respectively. The topography of projections from peripheral stimulation was approximately normal. Mossy fiber responses followed stimulation at up to 20/sec, whereas climbing fiber pathways fatigued at 10/sec. The latency of climbing fiber input to peripheral limb stimulation in x-irradiated cerebellum was 23 +/- 8 (SD) msec. In x-irradiated rats, the climbing fiber pathways evoked highly variable extracellular burst responses and intracellular EPSPs of different, discrete sizes. These variable responses suggest that multiple climbing fibers contact single Purkinje cells. We conclude that each type of afferent retains identifying characteristics of transmission. However, rules for synaptic specification appear to break down so that: (1) abnormal classes of neurons develop synaptic connections, i.e., mossy fibers to Purkinje cells; (2) incorrect numbers of neurons share postsynaptic targets, i.e., more than one climbing fiber to a Purkinje cell; and (3) inhibitory synaptic actions may be carried out in the absence of the major inhibitory interneurons, i.e., Purkinje cell collaterals may be effective in lieu of basket and stellate cells.     

Neural expectation: cerebellar and retinal analogs of cells fired by learnable or unlearned pattern classes

Neural networks are introduced which can be taught by classical or instrumental conditioning to fire in response to arbitrary learned classes of patterns. The filters of output cells are biased by presetting cells whose activation prepares the output cell to expect prescribed patterns. For example, an animal that learns to expect food in response to a lever press becomes frustrated if food does not follow the lever press. It's expectations are thereby modified, since frustration is negatively reinforcing. A neural analog with aspects of cerebellar circuitry is noted, including diffuse mossy fiber inputs feeding parallel fibers that end in Purkinje cell dendrites, climbing fiber inputs ending in Purkinje cell dendrites and giving off collaterals to nuclear cells, and inhibitory Purkinje cell outputs to nuclear cells. The networks are motivated by studying mechanisms of pattern discrimination that require no learning. The latter often use two successive layers of inhibition, analogous to horizontal and amacrine cell layers in vertebrate retinas. Cells exhibiting hue (in)constancy, brightness (in)constancy, or movement detection properties are included. These results are relevant to Land's retinex theory and to the existence of opponent- and nonopponent-type cell responses in retinal cells. Some adaptation mechanisms, and arousal mechanisms for crispening the pattern weights that can fire a given cell, are noted.Supported in part by the Alfred P. Sloan Foundation and the Office of Naval Research (N00014-67-A-0204-0051).     

Ratio of inferior olivary cells to Purkinje cells in a marsupial (Trichosurus vulpecula)

An indirect estimate of the extent of branching of the olivary axons in the cerebellum in a marsupial (Trichosurus vulpecula) was carried out. The cells in the inferior olivary nuclear complex (IOC) of both sides were estimated (mean = 57,200), as were the cerebellar Purkinje cells (mean = 881,300). Assuming that all climbing fibers arise from IOC cells and that each Purkinje cell receives a climbing fiber input, each IOC cell sends climbing fiber terminals to 15 Purkinje cells.     

Analysis of the dynamic behavior of neuron populations in the turtle cerebellum: II. Lumped circuit model

A lumped circuit model was constructed which consisted of two input channels, climbing fiber and mossy fiber afferents, which described the magnitudes of synaptic transmission and which accounted for synaptic and transmission delays. The parameters and coefficients of the transfer function were chosen such that they corresponded to physiological observable quantities. The corresponding time function approximated the data points. The results indicated that the dynamic behavior of the cerebellar circuit was satisfactorily accounted for by a parallel excitatory and inhibitory system with a combined climbing fiber and mossy-parallel fiber input exciting the Purkinje cells. The initial negative was predominantly a climbing fiber response of the Purkinje cell supporting the inference which was derived from purely electrophysiological data.     

Remodeling of monoplanar Purkinje cell dendrites during cerebellar circuit formation

Dendrite arborization patterns are critical determinants of neuronal connectivity and integration. Planar and highly branched dendrites of the cerebellar Purkinje cell receive specific topographical projections from two major afferent pathways; a single climbing fiber axon from the inferior olive that extend along Purkinje dendrites, and parallel fiber axons of granule cells that contact vertically to the plane of dendrites. It has been believed that murine Purkinje cell dendrites extend in a single parasagittal plane in the molecular layer after the cell polarity is determined during the early postnatal development. By three-dimensional confocal analysis of growing Purkinje cells, we observed that mouse Purkinje cells underwent dynamic dendritic remodeling during circuit maturation in the third postnatal week. After dendrites were polarized and flattened in the early second postnatal week, dendritic arbors gradually expanded in multiple sagittal planes in the molecular layer by intensive growth and branching by the third postnatal week. Dendrites then became confined to a single plane in the fourth postnatal week. Multiplanar Purkinje cells in the third week were often associated by ectopic climbing fibers innervating nearby Purkinje cells in distinct sagittal planes. The mature monoplanar arborization was disrupted in mutant mice with abnormal Purkinje cell connectivity and motor discoordination. The dendrite remodeling was also impaired by pharmacological disruption of normal afferent activity during the second or third postnatal week. Our results suggest that the monoplanar arborization of Purkinje cells is coupled with functional development of the cerebellar circuitry.     

Widespread state-dependent shifts in cerebellar activity in locomoting mice

Excitatory drive enters the cerebellum via mossy fibers, which activate granule cells, and climbing fibers, which activate Purkinje cell dendrites. Until now, the coordinated regulation of these pathways has gone unmonitored in spatially resolved neuronal ensembles, especially in awake animals. We imaged cerebellar activity using functional two-photon microscopy and extracellular recording in awake mice locomoting on an air-cushioned spherical treadmill. We recorded from putative granule cells, molecular layer interneurons, and Purkinje cell dendrites in zone A of lobule IV/V, representing sensation and movement from trunk and limbs. Locomotion was associated with widespread increased activity in granule cells and interneurons, consistent with an increase in mossy fiber drive. At the same time, dendrites of different Purkinje cells showed increased co-activation, reflecting increased synchrony of climbing fiber activity. In resting animals, aversive stimuli triggered increased activity in granule cells and interneurons, as well as increased Purkinje cell co-activation that was strongest for neighboring dendrites and decreased smoothly as a function of mediolateral distance. In contrast with anesthetized recordings, no 1-10 Hz oscillations in climbing fiber activity were evident. Once locomotion began, responses to external stimuli in all three cell types were strongly suppressed. Thus climbing and mossy fiber representations can shift together within a fraction of a second, reflecting in turn either movement-associated activity or external stimuli.     

A chondroitin sulfate proteoglycan that is developmentally regulated in the cerebellar mossy fiber system.

It is known that the mammalian brain contains many kinds of proteoglycans, but almost all of them remain to be characterized. In this study, we prepared a monoclonal antibody against a phosphate-buffered saline-soluble brain proteoglycan (MAb 6B4). MAb 6B4 recognized a 600- to 1000-kDa chondroitin sulfate proteoglycan with a 250-kDa core protein (6B4 proteoglycan). The core protein of 6B4 proteoglycan carried the HNK-1 epitope. Immunohistochemical analysis of the adult rat brain indicated that this proteoglycan was expressed on the cell surfaces of a subset of neurons. In the hindbrain, 6B4 proteoglycan was highly expressed on the cerebellar Purkinje cells and Golgi cells, and at particular nuclei including the pontine nuclei and lateral reticular nucleus. Almost all of these nuclei were connected to the cerebellum through the mossy fiber system. A developmental study indicated that the expression of this proteoglycan changed dramatically during the formation of the cerebellar mossy fiber system. The mossy fibers from the pontine nuclei expressed 6B4 proteoglycan transiently from Embryonic Day 20 (E20) to Postnatal Day 30 (P30), during which time the axonal outgrowth and glomerular synapse formation occurred. The Purkinje cells, glomeruli, and Golgi cells began to be stained with MAb 6B4 from P10, P16, and P20, respectively. These expression stages correspond with the onset of their synapse formation. These results suggest that 6B4 proteoglycan is closely involved in the development of the cerebellar mossy fiber system.     

Interrelated modification of excitatory and inhibitory synaptic connections in the olivary-cerebellar neuronal network

The model of simultaneous interrelated modification in the efficacy of synaptic inputs to different neurons of the olivary-cerebellar network is developed. The model is based on the following features of the network: simultaneous activation of the input layer (granule) cells and the output layer (deep cerebellar nuclei) cells by mossy fibers; simultaneous activation of Purkinje cells and cerebellar cells of the input and output layers by climbing fibers and their collaterals; the existence of local feedback excitatory, inhibitory, and disinhibitory circuits. The rise (decrease) of posttetanic Ca2+ concentration in reference to the level produced by previous stimulation causes the decrease (increase) in cGMP-dependent protein kinase G activity, and increase (decrease) inprotein phosphatase 1 activity. Subsequent dephosphorylation (phosphorylation) of ionotropic receptors results in simultaneous LTD (LTP) of the excitatory input together with the LTP (LTD) of the inhibitory input to the same neuron. The character of interrelated modifications of synapses at different cerebellar levels strongly depends on the olivary cell activity. In the presence (absence) of the signal from the inferior olive LTD (LTP) of the output cerebellar signal can be induced.     

A Mathematical Model of the Cerebellar-Olivary System I: Self-Regulating Equilibrium of Climbing Fiber Activity

We use a mathematical model to investigate how climbing fiber-dependent plasticity at granule cell to Purkinje cell (grPkj) synapses in the cerebellar cortex is influenced by the synaptic organization of the cerebellar-olivary system. Based on empirical studies, grPkj synapses are assumed to decrease in strength when active during a climbing fiber input (LTD) and increase in strength when active without a climbing fiber input (LTP). Results suggest that the inhibition of climbing fibers by cerebellar output combines with LTD/P to self-regulate spontaneous climbing fiber activity to an equilibrium level at which LTP and LTD balance and the expected net change in grPkj synaptic weights is zero. The synaptic weight vector is asymptotically confined to an equilibrium hyperplane defining the set of all possible combinations of synaptic weights consistent with climbing fiber equilibrium. Results also suggest restrictions on LTP/D at grPkj synapses required to produce synaptic weights that do not drift spontaneously.     

Climbing fiber innervation of NG2-expressing glia in the mammalian cerebellum

The molecular layer of the cerebellar cortex is populated by glial progenitors that express ionotropic glutamate receptors and extend numerous processes among Purkinje cell dendrites. Here, we show that release of glutamate from climbing fiber (CF) axons produces AMPA receptor currents with rapid kinetics in these NG2-immunoreactive glial cells (NG2+ cells) in cerebellar slices. NG2+ cells may receive up to 70 discrete inputs from one CF and, unlike mature Purkinje cells, are often innervated by multiple CFs. Paired Purkinje cell-NG2+ cell recordings show that one CF can innervate both cell types. CF boutons make direct synaptic junctions with NG2+ cell processes, indicating that this rapid neuron-glia signaling occurs at discrete sites rather than through ectopic release at CF-Purkinje cell synapses. This robust activation of Ca2+-permeable AMPA receptors in NG2+ cells expands the influence of the olivocerebellar projection to this abundant class of glial progenitors.     

Localization of Hexokinase in Neural Tissue: Electron Microscopic Studies of Rat Cerebellar Cortex

: The distribution of hexokinase (ATP:d -hexose 6-phosphotransferase, EC 2.7.1.1) in the rat cerebellar cortex has been studied at the electron microscopic level using the peroxidase-antiperoxidase procedure. Extensive staining of cytoplasmic regions, with some increased staining at mitochondrial profiles, was seen in the cell bodies of both neurons (basket, stellate, Lugaro, Golgi, and granule cells) and astrocytes. Oligodendrocytes showed little or no detectable staining. Purkinje cell perikarya were much less intensely stained than were the perikarya of other neurons. The initial portion of the Purkinje dendrite was, like the perikaryon from which it emerged, lightly stained. More intense staining was seen in the secondary and tertiary branches of the Purkinje dendrite, but the terminal branches were devoid of stain. Granule cell dendrites were well stained in their initial portions but devoid of stain in their terminal dendritic digits which form part of the cerebellar glomeruli. In contrast to the unstained granule cell dendritic digits, the central mossy fiber nerve terminal of the glomerulus exhibited intense staining of the mitochondrial profiles and of synaptic vesicles adjacent to the mitochondria. Axons of basket cells showed intense staining in the segments adjacent to the Purkinje cell soma, while terminal twigs of the basket axons in the pinceau surrounding the (unstained) initial segment of the Purkinje axon showed markedly decreased staining intensity. These results indicate that there may be substantial variation in hexokinase levels between the various regions of neuronal processes. Hexokinase was seen at both cytoplasmic and mitochondrial locations in a variety of cells. It does not appear likely that location of hexokinase can be directly correlated with cell type, i.e., with neurons versus glia.     

Electron microscopical study of synaptic glomeruli in cerebellum transplanted to the anterior eye chamber

Fetal cerebellar anlage from rat fetuses of 15-16 operational days were grafted into the anterior chamber of the eye of adult female albino rat recipients. Survival time of the transplants--containing both cerebellar cortex and cerebellar nuclei--was 2 to 2 1/2 months. Electron microscopical (EM) studies of the thin, under-developed granular layer of the laminated cerebellar cortex revealed the presence of well differentiated cerebellar glomeruli, surrounded by granule cell perikarya. As in the normal cerebellar cortex, the central profile of the glomerular complex was the large mossy terminal, containing spheroid synaptic vesicles, and forming synaptic contacts with dendrites and dendritic digits of the granule cells. Golgi cell axonal varicosities, containing ovoid or pleomorphic synaptic vesicles were found also on the periphery of the glomeruli. In addition, in several synaptic glomeruli, a third neuronal element was also observed, containing flat, discoidal vesicles and receiving synaptic contacts from mossy and Golgi axons, but being also presynaptic to granule cell dendrites. It is suggested that all mossy terminals in the cerebellar transplant originate from the cerebellar nucleus. Morphological evidence is also provided that the presynaptic dendrite-like processes--never found in normal cerebellar cortex--are also processes of nuclear neurons.     

Hog cerebellar D-amino acid oxidase and its histochemical and immunofluorescent localization.

Abstract— Hog cerebellar d -amino acid oxidase (d -AAO; EC 1.4.3.3) has been purified to homogeneity. The enzyme was found to be indistinguishable from crystalline hog kidney d -AAO by a number of criteria, including electrophoresis in both cationic and anionic discontinuous buffer systems, FAD and sulfhydryl contents, and monomer molecular weights of about 40,000 as determined by SDS disc gel electrophoresis. Both preparations exhibited similar specific activities (23 μmol d -ala oxidized min^?1 mg^?1 protein), substrate specificities, and susceptibilities to competitive inhibitors. Rabbit antisera were prepared against each enzyme preparation. Double immunodiffusion revealed no antigenic differences between the two when antiserum against either preparation was used. Although a soluble protein, d -AAO activity in cerebellum is particulate. Two methods were utilized to study the histological localization of d -AAO activity in hog cerebellum: peroxidase-coupled histochemistry and immunofluorescence. The histochemical procedure seems specific for d -AAO since l -amino acids are inert and known competitive inhibitors of the purified flavoenzyme prevent staining. Rabbit antiserum prepared against purified hog cerebellar d -AAO was visualized indirectly with fluorescein-labeled goat antirabbit IgG antiserum. Control experiments with serum from unimmunized rabbits were negative. The results of both techniques were identical in three respects: (1) d -AAO was observed in many fibers emanating from cerebellar white matter; the white matter itself did not exhibit d -AAO, despite presence of the oxidase by biochemical assay; (2) intense d -AAO activity (or antigen) was found in mossy fiber rosettes (glomeruli) in the granular layer; (3) a peculiar and intense localization of d -AAO was noted at the level of, but not within, the Purkinje cell soma. The molecular layer was essentially devoid of d -AAO histochemical activity except for minimal staining near the pial surface. However, the more sensitive immunofluorescent technique revealed d -AAO containing fibers in the molecular layer running parallel to each other and perpendicular to the pial surface; the relatively large size and small number of these fibers do not suggest identification as granule cell axons. No d -AAO has been found in granule cell soma, Golgi Type-II cells, or Purkinje cell soma. These results are discussed in terms of the localization of d -AAO in mossy fibers and their terminals and in certain cell-type(s) of cerebellar origin.     

Development and evolution of cerebellar neural circuits

The cerebellum controls smooth and skillful movements and it is also involved in higher cognitive and emotional functions. The cerebellum is derived from the dorsal part of the anterior hindbrain and contains two groups of cerebellar neurons: glutamatergic and gamma-aminobutyric acid (GABA)ergic neurons. Purkinje cells are GABAergic and granule cells are glutamatergic. Granule and Purkinje cells receive input from outside of the cerebellum from mossy and climbing fibers. Genetic analysis of mice and zebrafish has revealed genetic cascades that control the development of the cerebellum and cerebellar neural circuits. During early neurogenesis, rostrocaudal patterning by intrinsic and extrinsic factors, such as Otx2, Gbx2 and Fgf8, plays an important role in the positioning and formation of the cerebellar primordium. The cerebellar glutamatergic neurons are derived from progenitors in the cerebellar rhombic lip, which express the proneural gene Atoh1. The GABAergic neurons are derived from progenitors in the ventricular zone, which express the proneural gene Ptf1a. The mossy and climbing fiber neurons originate from progenitors in the hindbrain rhombic lip that express Atoh1 or Ptf1a. Purkinje cells exhibit mediolateral compartmentalization determined on the birthdate of Purkinje cells, and linked to the precise neural circuitry formation. Recent studies have shown that anatomy and development of the cerebellum is conserved between mammals and bony fish (teleost species). In this review, we describe the development of cerebellar neurons and neural circuitry, and discuss their evolution by comparing developmental processes of mammalian and teleost cerebellum.     

剌激中缝背核压抑大鼠小脑浦肯野细胞对苔状...

In anaesthetized and paralyzed rats, the effect of dorsal raphe (DR) conditioning stimulation on cerebellar Purkinje cell (PC) responses to mossy fiber and climbing fiber inputs were examined. The main results are as follows: (1) Stimulation of cerebral sensorimotor cortex elicits widespread activation of mossy and climbing fiber inputs to PCs in contralateral VI and VII lobules of the cerebellum and generates two kinds of evoked responses, i.e. the simple spike (SS) and the complex spike (CS) responses with respectively a latency 8-25 and 12-30 ms. (2) These PC responses could be markedly suppressed by stimulation of DR at intensities which by themselves were subthreshold for directly affecting PC's spontaneous SS and CS activities. (3) This DR-induced depressive effects on evoked PC's SS and CS excitations could be attenuated or blocked by systemic administration of 5-HT receptor blocker methysergide. These results demonstrate that serotonergic fiber input from DR can suppress the efficacy of mossy and climbing fiber synaptic action on PC, or decrease the responsiveness of PC itself to afferent synaptic action. The findings of this study also suggest that the raphe-cerebellar serotonergic fiber afferent system may be involved in some of the important neuronal processing in the cerebellum.