Observations on the ultrastructure of the developing cerebellum of the Macaca mulatta

The fine structure of the cerebellar cortex of the macaque fetus was examined at 75, 100, 125, 150, and 175 days after conception. Young Purkinje cells were recognizable in the 75 and 100-day old fetuses, occurring in clusters several layers deep. The rough endoplasmic reticulum in the Purkinje cells was limited to the perinuclear area and the region of apical cytoplasm adjacent to the plasmalemma. Neither dendritic processes nor synaptic contacts were seen on Purkinje cells at this time, although an occasional observation was made of synapses involving other cell types. Between 100 and 125 days, the cerebellar cortex matured rapidly. At the latter time, the Purkinje cells had a well developed dendrite, and axodendritic and axosomatic synapses were seen. At 150 days gestation age the Purkinje cells had well developed Nissl substance, and the axodendritic synapses were similar in structure to the adult spine contact. The relationship between maturation of the thyroid gland and of the cerebellum in the macaque is discussed.     

Synaptic 5'-nucleotidase is transient and indicative of climbing fiber plasticity during the postnatal development of rat cerebellum

The transient appearance of 5'-nucleotidase, an adenosine-producing ecto-enzyme, was studied during specific stages of postnatal synaptogenesis in the rat cerebellum. For ultrastructural detection of 5'-nucleotidase activity, an enzyme-cytochemical technique was used. Between postnatal days 4 and 6, enzymatic reaction product was present in the synaptic clefts of climbing fibers containing the perisomatic spines, apical cones and emerging dendrites of Purkinje cells (CF-PC synapses). Labeled parallel fiber synapses were observed on dendritic shafts of cerebellar interneurons. At postnatal days 9 and 12, enzyme-positive parallel fiber terminals were in addition numerous on the spines of peripheral Purkinje branchlets, and gradually disappeared thereafter. Between postnatal days 8 and 15, labeling of perisomatic CF-PC contacts persisted. In contrast, climbing fiber synapses on Purkinje dendrites were only occasionally labeled. Between postnatal days 18 and 21, synaptic reaction product was restricted to mossy fibers. At the same time, association of 5'-nucleotidase with glial profiles was prominent throughout the cerebellar layers. In adult cerebellum (from 24 days onwards) all synapses were devoid of enzymatic activity. Throughout development, basket, stellate and Golgi cell synapses were devoid of enzymatic activity. We conclude that 5'-nucleotidase is present in excitatory cerebellar synapses during part of their generation period. The transient nature of this phenomenon suggests that 5'-nucleotidase may serve as a novel, cytochemical marker for a specific state of synaptic maturation, and in particular for climbing fiber plasticity. A role of 5'-nucleotidase in purinergic neuromodulation and cellular contact formation could be significant in these processes.     

The cerebellum of the frog Rana ridibunda. An electron microscopic study

An electron microscopic study of neuronal types and different synaptic contacts has been made in the cerebellum of the frog Rana ridibunda. The Purkinje cells have a pear-shaped cell body and in their cytoplasm the organelles show a special arrangement because of the great amount of microtubules they contain. The granule cells are small, rounded neurons with a large nucleus surrounded by a thin rim of cytoplasm. The stellate cells are interneurons of the molecular layer whose large nuclei show a single finger-like invagination of its nuclear envelope. The afferent tracts to the cerebellum end either as climbing fibers or mossy fibers. The axon terminals of climbing fibers are large and the synaptic complexes exhibit all the features of a type-I Gray synapse. The mossy fibers reach the granular layer and synapses between them and granule cell dendrites are by far the most abundant. The parallel fibers establish synaptic contacts on the spines arising from the spiny branchlet units of the Purkinje cells and with the perikaryon and dendrites of stellate cells. The stellate cell axons cross the molecular layer and establish type-II Gray synapses on the Purkinje cells.     

A Golgi study of cerebellar malformation in 13 trisomy

The structure of cerebellar malformations in the brains of two infants with 13 trisomy has been studied by means of the Golgi method. Poorly organized cerebellar dysplasias (heterotaxias) are composed of Purkinje, Golgi and granule cells arranged and oriented in a disorderly fashion. The variable orientation and organization of the dendritic arbor of Purkinje cells within these cellular aggregates is supposed to be related to abnormal distribution of parallel fibers. Large ganglion cell heterotopias are not a homogeneous group, but two distinct types may be defined. First, Purkinje cell heterotopias which are located in the white matter of the cerebellum below the normally formed folia; these are composed of large neurons with arrested migration to the cortex. Secondly, multipolar cell heterotopias which are located in the deep white matter near the dentate and the roof nuclei, formed of neurons belonging to the deep cerebellar nuclei.     

Morphological development of dendritic spines on rat cerebellar Purkinje cells

The posterior cerebellum is strongly involved in motor coordination and its maturation parallels the development of motor control. Climbing and mossy fibers from the spinal cord and inferior olivary complex, respectively, provide excitatory afferents to cerebellar Purkinje neurons. From post-natal day 19 climbing fibers form synapses with thorn-like spines located on the lower primary and secondary dendrites of Purkinje cells. By contrast, mossy fibers transmit synaptic information to Purkinje cells trans-synaptically through granule cells. This communication occurs via excitatory synapses between the parallel fibers of granule cells and spines on the upper dendritic branchlets of Purkinje neurons that are first evident at post-natal day 21. Dendritic spines influence the transmission of synaptic information through plastic changes in their distribution, density and geometric shape, which may be related to cerebellar maturation. Thus, spine density and shape was studied in the upper dendritic branchlets of rat Purkinje cells, at post-natal days 21, 30 and 90. At 90 days the number of thin, mushroom and thorn-like spines was greater than at 21 and 30 days, while the filopodia, stubby and wide spines diminished. Thin and mushroom spines are associated with increased synaptic strength, suggesting more efficient transmission of synaptic impulses than stubby or wide spines. Hence, the changes found suggest that the development of motor control may be closely linked to the distinct developmental patterns of dendritic spines on Purkinje cells, which has important implications for future studies of cerebellar dysfunctions.     

Synaptic organization of the mouse cerebellar cortex in organotypic slice cultures

The cellular and synaptic organization of new born mouse cerebellum maintained in organotypic slice cultures was investigated using immunohistochemical and patch-clamp recording approaches. The histological organization of the cultures shared many features with that observed in situ. Purkinje cells were generally arranged in a monolayer surrounded by a molecular-like neuropil made of Purkinje cell dendritic arborizations. Purkinje cell axons ran between clusters of small round cells identified as granule cells by Kv3.1b potassium channel immunolabelling. The terminal varicosities of the Purkinje cells axons enwrapped presumptive neurons of the cerebellar nuclei whereas their recurrent collaterals were in contact with Purkinje cells and other neurons. Granule cell axons established contacts with Purkinje cell somata and dendrites. Parvalbumin and glutamine acid decarboxylase (GAD) immunohistochemistry revealed the presence of presumptive interneurons throughout the culture. The endings of granule cell axons were observed to be in contact with these interneurons. Similarly, interneurons endings were seen close to Purkinje cells and granule cells. Whole cell recordings from Purkinje cell somata showed AMPA receptor-mediated spontaneous excitatory post-synaptic currents (sEPSCs) and GABAA receptor-mediated spontaneous inhibitory post-synaptic currents (sIPSCs). Similar events were recorded from granule cell somata except that in this neuronal type EPSPs have both a NMDA component and an AMPA component. In addition, pharmacological experiments demonstrated a GABAergic control of granule cell activity and a glutamatergic control of GABAergic neurons by granule cells. This study shows that a functional neuronal network is established in such organotypic cultures even in the absence of the two normal excitatory afferents, the mossy fibers and the climbing fibers.     

The Purkinje neuron: II. Electron microscopic analysis of the mature Purkinje neuron in organotypic culture

Purkinje neurons of organotypic cultures were investigated electron microscopically following their analysis with the Golgi technique. The purpose of this study was to critically examine the issue of synaptic specificity in CNS cultures. The unique finding was the synaptic cluster, a terminal which engulfs many Purkinje spines. In the neuropil and on the major dendrites, this synaptic arrangement was interpreted to be a hypertrophic parallel fiber, representing a type of synaptic modulation. The terminals on the somatic spines are also in the form of clusters; some or all of these spines were thought to have developed to form synapses with the climbing fiber. In the absence of this afferent, the parallel fiber—a competing system—takes over the site. This represents a form of synaptic plasticity in these cultures. The inhibitory synaptic relationships were maintained on the soma and dendrites, but it was found that the basket synapses did not quantitatively encase the soma as is seen in the intact animal. Mossy-type terminals were found occasionally synapsing with Purkinje dendritic spines, as has been seen in agranular cerebellum in animals. These mossy terminals are presently thought to originate from the deep cerebellar nuclei within these cultures. Synaptic errors were rarely encountered. It is concluded that this preparation develops in accordance with established neurobiological principles, that the Purkinje neuron reaches a mature state in culture, and that this model has a sound anatomical basis for further experimental work.     

Cerebellar efferent neurons in teleost fish

In tetrapods, cerebellar efferent systems are mainly mediated via the cerebellar nuclei. In teleosts, the cerebellum lacks cerebellar nuclei. Instead, the cerebellar efferent neurons, termed eurydendroid cells, are arrayed within and below the ganglionic layer. Tracer injections outside of the cerebellum, which retrogradely label eurydendroid cells demonstrate that most eurydendroid cells possess two or more primary dendrites which extend broadly into the molecular layer. Some eurydendroid cells mostly situated in caudal portions of the cerebellum have only one primary dendrite. The eurydendroid cells receive inputs from the Purkinje cells and parallel fibers, but apparently do not receive inputs from the climbing fibers. Eurydendroid cells of the corpus cerebelli and medial valvula project to many brain regions, from the diencephalon to the caudal medulla. A few eurydendroid cells in the valvula project directly to the telencephalon. About half of the eurydendroid cells are aspartate immunopositive. Anti-GABA and anti-zebrin II antibodies that are known as markers for the Purkinje cells in mammals also recognize the Purkinje cells in the teleost cerebellum, but do not recognize the eurydendroid cells. These results suggest that the eurydendroid cells receive GABAergic inputs from the Purkinje cells. This relationship between the eurydendroid and Purkinje cells is similar to that between the cerebellar nuclei and Purkinje cells in mammals. The eurydendroid cells of teleost have both dissimilar as well as similar features compared to neurons of the cerebellar nuclei in tetrapods.     

Neurogenesis of the climbing fibers in the human cerebellum: a Golgi study

The prenatal and early postnatal neurogenesis of the human climbing fibers of the lateral cerebellar hemispheres have been studied, with the rapid Golgi method, and correlated with the developmental stages of Purkinje cells. A transitional phase has been established in the neurogenesis of the human Purkinje cell between the second and third stages of Cajal. This phase coincides with the arrival of the climbing fibers. It is characterized by the reabsorption and subsequent transformation of Purkinje cell's basal dendrites into somatic spines. Following the arrival of the climbing fibers and the establishment of contacts, the Purkinje cell is progressively transformed from an immature stellate and nonoriented cell into a monopolar and spatially oriented one which acquires all of its mature morphological and functional features. The human climbing fibers arrive at the Purkinje cell plate by the 28th week of gestation and establish a transient paraganglionic plexus before contacts with these neurons can be recognized. They start to form pericellular nests by the 29th week, and by the 31st week of gestation all Purkinje cells of the lateral hemispheres have pericellular nests around their bodies. These pericellular nests are progressively and rapidly transformed into supracellular "capuchones" which themselves are also short-lived because the climbing process starts readily in them. Supracellular "capuchones" are recognized by the 34th seek and their fibrils start to climb the dendrites of Purkinje cells (young climbing phase) by the 36th week of gestation. The process of climbing the dendrites of the Purkinje cells will continue through late prenatal and early postnatal life. The human climbing fibers are distributed, in the internal granular layer, within narrow and long vertical territories which are transverse to the long axis of the follium. A single climbing fiber is (1) able to establish contacts with many Purkinje cells located within its narrow territory of distribution; (2) has a tendency to establish contacts with small groups of Purkinje cells rather than with isolate neurons; (3) able to send collaterals to several contiguous cerebellar folia; and (4) able to send collaterals to the internal granular layer and to form pericellular nests in it. The human cerebellum may be considered to be subdivided into a series of parallel, narrow, and transverse structural/functional planes, each one characterized by the distribution of a climbing fiber.(ABSTRACT TRUNCATED AT 400 WORDS)     

Palisade pattern of mormyrid Purkinje cells: a correlated light and electron microscopic study

The present study is devoted to a detailed analysis of the structural and synaptic organization of mormyrid Purkinje cells in order to evaluate the possible functional significance of their dendritic palisade pattern. For this purpose, the properties of Golgi-impregnated as well as unimpregnated Purkinje cells in lobe C1 and C3 of the cerebellum of Gnathonemus petersii were light and electron microscopically analyzed, quantified, reconstructed, and mutually compared. Special attention was paid to the degree of regularity of their dendritic trees, their relations with Bergmann glia, and the distribution and numerical properties of their synaptic connections with parallel fibers, stellate cells, "climbing" fibers, and Purkinje axonal boutons. The highest degree of palisade specialization was encountered in lobe C1, where Purkinje cells have on average 50 palisade dendrites with a very regular distribution in a sagittal plane. Their spine density decreases from superficial to deep (from 14 to 6 per micron dendritic length), a gradient correlated with a decreasing parallel fiber density but an increasing parallel fiber diameter. Each Purkinje cell makes on average 75,000 synaptic contacts with parallel fibers, some of which are rather coarse (0.45 microns), and provided with numerous short collaterals. Climbing fibers do not climb, since their synaptic contacts are restricted to the ganglionic layer (i.e., the layer of Purkinje and eurydendroid projection cells), where they make about 130 synaptic contacts per cell with 2 or 3 clusters of thorns on the proximal dendrites. These clusters contain also a type of "shunting" elements that make desmosome-like junctions with both the climbing fiber boutons and the necks of the thorns. The axons of Purkinje cells in lobe C1 make small terminal arborizations, with about 20 boutons, that may be substantially (up to 500 microns) displaced rostrally or caudally with respect to the soma. Purkinje axonal boutons were observed to make synaptic contacts with eurydendroid projection cells and with the proximal dendritic and somatic receptive surface of Purkinje cells, where about 15 randomly distributed boutons per neuron occur. The organization of Purkinje cells in lobe C3 differs markedly from that in C1 and seems to be less regular and specialized, although the overall palisade pattern is even more regular than in lobe C1 because of the absence of large eurydendroid neurons. However, individual neurons have a less regular dendritic tree, there is no apical-basal gradient in spine density or parallel fiber density and diameter, and there are no "shunting" elements in the climbing fiber glomeruli.(ABSTRACT TRUNCATED AT 400 WORDS)     

Development of cholinesterase histochemical staining in cerebellar cortex: transient expression of "nonspecific" cholinesterase in Purkinje cells of the nodulus and uvula.

Patterns of "nonspecific" cholinesterase (ChE) and acetylcholinesterase (AChE) activity were studied in developing rat cerebellar cortex by enzyme histochemistry and light and electron microscopy. Three types of ChE histochemical reaction product were observed in cerebellar cortex: (i) ChE is found in capillary endothelium throughout the cerebellum. Capillary ChE staining is present by the time of birth and continues into adulthood. (ii) ChE is found in radial glial fibers and their parent cell bodies, the Golgi epithelial cells. Radial glial fiber staining is mot intense during the first 3 weeks of postnatal life. (iii) ChE is found in Purkinje cells of the nodulus and ventral uvula. No ChE staining of Purkinje cells was seen in other parts of the cerebellum. ChE staining of Purkinje cells appears to be transient, first appearing at Postnatal Day 2 (P2), reaching peak intensity at P7-9, and decreasing to adult levels by P16. AChE activity displays a pattern markedly different from ChE, with staining in deep cerebellar nuclei, in putative mossy fiber terminals, and in Golgi neurons of cerebellar cortex. No evidence was found for transient AChE staining in Purkinje cells in any part of the cerebellum. The function of transiently expressed ChE activity in developing Purkinje neurons is unknown, but may be related to reorganization of cerebellar cortical circuitry associated with growth of mossy fiber afferents.     

THE EFFECTS OF CARBON MONOXIDE ON NEURONS OF THE CEREBELLUM*

The effects of carbon monoxide on neurons of the cerebellum Acute CO exposure in rats produces multifocal edema and neuroglial swelling in cerebellar cortex and nuclei. Chronic multiple daily exposures to CO produce degenerative changes in neurons–shrinkage of cell somata and dendrites, enlargement of nuclei and nucleoli, vacuolation in neuronal cytoplasm, pyknosis and ultimate engulfment by microglia. Focal areas of Purkinje cell loss are typical; individual basket cells and their axons, stellate cells and the deep nuclear neurons are severely affected. Counts of Purkinje cells and their dendritic thorns show reductions of 30% and 31% respectively in affected regions. In foci of severe Purkinje cell loss, dendritic thorns of the neighboring cells which remain assume a greater number of synapses with unusual presynaptic partners, e. g. both parallel fibers and climbing fibers in a single thorn. An increase of 4.2% more synapses per thorn was measured when compared to the normal. Thus, the stress of chronic CO exposure stimulates synaptic plasticity or a compensatory remodelling of neuronal elements in the central nervous system. A scheme is proposed to summarize these changing relationships caused by the loss of some dendritic thorns and the migration of the presynaptic partners to other surviving thorns.     

Organizational principles and microcircuitry of the cerebellum

The cerebellum is known to influence motor behavior and to enable smooth, coordinated movements. Recent evidence also suggests that the cerebellum contributes to non-motor behavior, including components of cognition and regulation of affective state.This review summarizes the organization and circuitry of the cerebellum as a basis for understanding newly emerging concepts about the function of this neuronal system. The cerebellum consists of several divisions with separate functions. One region is associated with the vestibular system and another with brainstem and spinal cord. A third region, the cerebrocerebellum, has extensive interconnections with cerebral cortex and is likely to be involved in motor coordination and regulation of non-motor behavior. The cerebellar cortex is made up of radial modules of interconnected neurons. The Purkinje cell is the principle integrating neuron and focal point of each module. Other neuron types include the granule cell and three inhibitory interneurons. The Purkinje cell integrates excitatory inputs from climbing and parallel fibers, while its axon modulates activity of neurons in the deep nuclei, which represents the final outflow from cerebellum to other parts of the brain. Cerebellar circuitry exhibits a strong parasagittal organization based on climbing fiber input and the distributions of neuronal proteins and neuronal vulnerability to insults. The combination of this parasagittal circuitry with the mediolateral course of parallel fibers results in a Cartesian coordinate system which is likely to be a crucial factor in its signal processing function. Although numerous details of cerebellar microcircuitry, synaptic transmission and signal transduction have been determined, the functional contribution of cerebellar signalling to brain function remains highly enigmatic.     

Purkinje cell neuroaxonal dystrophy similar to nervous mutant mice phenotype in two sibling kittens

Three 4-month-old kittens from the same litter were presented, two of which were exhibiting cerebellar signs. Euthanasia was requested. No cerebellum atrophy was disclosed on necropsy. General cerebellar anatomy was normal, including the thickness of the cortical layers, myelination, and neurons of the deep cerebellar nuclei. In the ataxic cat vermis, Purkinje cells were lacking along broad parasagittal bands symmetrically disposed relative to the midline. Many Purkinje cells were also lacking in the hemispheres. The nodulus and the flocculus were normal. Surviving Purkinje cells had frequent main dendrite swellings visible with anti-calbindin and anti-microtubule associated protein. In affected regions, calbindin and phosphorylated neurofilaments immunesera stained numerous axonal torpedoes located in the granular layer and the folial white matter. They were also present in processes of the deep cerebellar nuclei and lateral vestibular nucleus. Loss of synaptic endings onto the neurons of these nuclei was evident. Hypertrophied Purkinje cell recurrent axons and enhanced retrograde synaptic endings were present in the granular layer. Bergmann glia was strongly labeled by anti-GFAP, but no abnormal supplementary fibers were seen. None of these alterations were present in the normal sister. However, abnormal vacuolation of the Purkinje cell main dendrites was evident in all three cats, but not in six unrelated control cats that were 3–6 months old. The inferior olive and pontine nuclei were also normal. The two ataxic cats had a primary Purkinje cell degeneration that shared many common features with the abnormal Purkinje cells of the nervous mutant mouse.     

Postsynaptic membrane domains in the molecular layer of the cerebellum: a correlation between presynaptic inputs and postsynaptic plasma membrane organization

The intramembrane particle (IMP) content of Purkinje, basket, stellate and Golgi cell plasma membrane was quantitatively assessed in freeze-fracture replicas of the cerebellum of normal rats and Weaver mutant mice. This analysis showed that, irrespective of the cell type innervated (i.e. Purkinje, stellate, basket or Golgi cells) postsynaptic membranes for parallel fibers had a relatively low IMP content in their cytoplasmic P-face (≈ 750 IMP/μm², while postsynaptic membranes for climbing, basket and stellate axons were characterized by a significantly higher IMP content (≈ 1400 IMP/μm²). This difference of IMP content between the targets for parallel fibers and those for climbing, basket and stellate axons was restricted to 3IMP smaller than 10 nm and appeared progressively during the development of the molecular layer, suggesting a correlation between the formation of synaptic contacts and the segregation of the postsynaptic membrane in these two different domains. In addition, the study of the Weaver mice cerebellum, which is deprived of parallel fibers, but yet shows a normal IMP content in the postsynaptic membrane for the missing fibers, indicated that this characteristic IMP content is established before or during the afferent's reaching its target, and independently of whether the contact ultimately occurs.     

Activity-dependent axonal and synaptic plasticity in the cerebellum

The cerebellum is a brain region endowed with a high degree of plasticity also in adulthood. After damage or alteration in the patterns of activity, it is able to undergo remarkable changes in its architecture and to form new connections based upon a process of synaptic reorganization. This review addresses cellular and molecular mechanisms that regulate the competition between two inputs belonging to different neuronal populations in innervating two contiguous but separate domains of the same target cell. The two inputs are the parallel fibers, the axon of the cerebellar granule cells, and the olivocerebellar neurons, that terminate as climbing fibers in the cerebellar cortex. The target is the Purkinje cell characterized by two dendritic domains that are different in size and number of spines, upon which the two afferent inputs impinge. Both inputs express several genes related to plasticity throughout the life span conferring the ability to remodel their synapses. In addition, we provided evidence that climbing fibers and Purkinje cells show remarkable reciprocal trophic interactions that are required for the maintenance of the correct synaptic connectivity. Through their activity, climbing fibers sustain the competition with parallel fibers by displacing this input to the distal territory of the Purkinje cell dendrite. In addition, they operate on the Purkinje cells through AMPA receptor suppressing spines in the territory surrounding their synapses. In this way, climbing fibers are able to optimize spine distribution and functional connectivity.     

Parallel fiber receptive fields: a key to understanding cerebellar operation and learning

In several theories of the function of the cerebellum in motor control, the mossy-fiber-parallel fiber input has been suggested to provide information used in the control of ongoing movements whereas the role of climbing fibers is to induce plastic changes of parallel fiber (PF) synapses on Purkinje cells. From studies of climbing fibers during the last few decades, we have gained detailed knowledge about the zonal and microzonal organization of the cerebellar cortex and the information carried by climbing fibers. However, properties of the PF input to Purkinje cells and inhibitory interneurones have been largely unknown. The present review, which focuses on the C3 zone of the cerebellar anterior lobe, will present and discuss recent data of the cutaneous PF input to Purkinje cells, interneurons and Golgi cells as well as novel forms of PF plasticity.     

Early climbing fiber interactions with Purkinje cells in the postnatal mouse cerebellum.

The time and place of initial contacts between afferent axons and their target cells are not known for most regions of the mammalian CNS. To address this issue, we have selectively visualized afferent climbing fiber axons together with their synaptic targets, Purkinje cells, in postnatal mouse cerebellum. Climbing fibers were orthogradely labeled by injection of rhodamine isothiocyanate into their brainstem source, the inferior olivary nucleus. Purkinje cells were localized with an antibody to a calcium-binding protein, calbindin D-28k (CaBP), in the same section or in adjacent sections. A novel view of the olivocerebellar projection and the morphology of climbing fiber arbors prior to the well-known "nest" stage has emerged from this analysis. At birth, climbing fibers project into the zone of Purkinje cells, before these cells have aligned into a monolayer. During this phase, climbing fibers have simple morphologies consisting of relatively unbranched terminal arbors and small tapered growing tips. Purkinje cells are arranged 3-6 cells deep and have tufted dendrites and relatively smooth somata. By postnatal days 3-4, climbing fibers branch over several adjacent Purkinje cell perikarya, which are still organized in a band several cells thick. From postnatal days 5-7, when climbing fibers subsequently make focused nests on individual cells, Purkinje somata are smoother and form a more distinct monolayer. Up to this time, however, climbing fibers continue to associate with Purkinje perikarya, even though Purkinje cell dendrites have emerged and branched extensively. By postnatal days 8-10, climbing fiber terminals climb onto the trunk of the relatively mature Purkinje dendritic tree. At birth, mossy fibers originating from the pontine nuclei resemble immature climbing fibers in that they also have a simple unbranched morphology and growing tips, but project only so far as the internal granule cell layer. Occasional individual fibers reach into the Purkinje zone both at postnatal day 0 and postnatal day 4, confirming that the fibers formerly described as "combination fibers" (Mason and Gregory, S4. J. Neurosci, 4:1715-1735) can be mossy in origin. These data demonstrate that climbing fibers project among Purkinje cells earlier than suspected, before these afferents begin to arborize and form pericellular nests. Our observations are not in accord with the view derived from autoradiographic tracing studies that as in other cortical areas, climbing afferents wait in the vicinity of Purkinje cells in the early neonatal period, then advance onto these cells in synchrony with Purkinje cell alignment into a monolayer and dendritic maturation.(ABSTRACT TRUNCATED AT 400 WORDS)