Copula Pyramidis Research Articles

Projections from the lowest thoracic and upper lumbar segments to the cerebellar cortex in the rat: An anterograde tracing study

The projections from the lowest thoracic and upper lumbar (T13 to L3) segments to the cerebellar cortex were examined by anterograde tracing with biotinylated dextran amine in the rat. Unilateral injections resulted in bilateral labeling of mossy fiber terminals in lobules Ib to VI, VIII, IX and copula pyramidis. The majority (64 % of the total 30,526 labeled terminals) were present ipsilaterally in lobules II (8.5 %), III (20 %), IV (11 %), V (12 %), VIII (4.1 %), and copula pyramidis (6.8 %). The projection field in the anterior lobe was composed of five longitudinal areas: area 1 in the midline region, areas 2 and 3 in the middle and lateral parts of the vermis, and areas 4 and 5 in the medial part of the intermediate region of the hemisphere. The projection areas are characteristically localized in the apical to the middle part of the lobule. In the posterior lobe, the longitudinal areas were present in the midline region, the middle and lateral parts of lobules VIIIa and VIIIb, and the medial and middle parts of copula pyramidis. The present study reveals the whole areas and the pattern of projections from the segments containing the cells of origin of the dorsal and ventral spinocerebellar tracts.

Reorganization of longitudinal compartments in the laterally protruding paraflocculus of the postnatal mouse cerebellum.

The paraflocculus and the neighboring smaller flocculus form a remarkable protrusion in the ventrolateral aspect of the mouse cerebellum, in which the longitudinal compartments are conspicuously oriented perpendicularly to the sagittal plane. The developmental process of such anatomical arrangements in these lobules has not been fully clarified. Here, we used the genetic tractability of pcdh10-lacZ knock-in (OL-KO), IP 3 R1-nls-lacZ transgenic (1NM13) and Gpr26cre-Ai9-AldocV mice to track the development of compartments and examined local longitudinal orientation of Purkinje cells within the paraflocculus and flocculus. We observed a distinct pcdh10-positive (pcdh10+) compartment in the flocculus, whereas the paraflocculus and other lobules had a continuous paravermal pcdh10+ compartment, in the embryonic OL-KO cerebellum. During the first postnatal week, the parafloccular pcdh10+ compartment shifted laterally to the most lateral edge in the caudal part of the protruding paraflocculus. Although the most medial edge of the parafloccular pcdh10+ compartment remained in the nonprotruding part of the paraflocculus, it was disrupted from the originally continuous pcdh10+ compartment in the copula pyramidis. The local longitudinal orientation changed gradually along with the mediolateral extent of the copula pyramidis, almost becoming perpendicular to the sagittal plane in the laterally connected paraflocculus in the adult cerebellum. This rotational change in orientation was derived from the short U-shaped embryonic cerebellum, in which the surfaces of the flocculus and paraflocculus were oriented laterally. These results indicated that the peculiar compartmental organization of the paraflocculus originates from the embryonic common hemispheric compartmental organization and shaped by the significant reorganization process in the first postnatal week.

Divergent projections of single pontocerebellar axons to multiple cerebellar lobules in the mouse

The basilar pontine nucleus (PN) is the key relay point for the cerebrocerebellar link. However, the projection pattern of pontocerebellar mossy fiber axons, which is essential in determining the functional organization of the cerebellar cortex, has not been fully clarified. We reconstructed the entire trajectory of 25 single pontocerebellar mossy fiber axons labeled by localized injection of biotinylated dextran amine into various locations in the PN and mapped all their terminals in an unfolded scheme of the cerebellum in 10 mice. The majority of axons (20/25 axons) entered the cerebellum through the middle cerebellar peduncle contralateral to the origin, while others entered through the ipsilateral pathway. A small number of axons (1/25 axons) had collaterals terminating in the cerebellar nuclei. Axons projected mostly to a combination of lobules, often bilaterally, and terminated in multiple zebrin (aldolase C) stripes, more frequently in zebrin-positive stripes (83.9%) than in zebrin-negative stripes, with 66.5 mossy fiber terminals on the average. Axons originating from the rostral (plus medial and lateral), central and caudal PN mainly terminated in the paraflocculus, crus I and lobule VIb-c, in the simplex lobule, crus II and paramedian lobule, and in lobules II-VIa, VIII and copula pyramidis, respectively. The results suggest that the interlobular branching pattern of pontocerebellar axons determines the group of cerebellar lobules that are involved in a related functional localization of the cerebellum. In the hemisphere, crus I may be functionally distinct from neighboring lobules (simple lobule and crus II) in the mouse cerebellum based on the pontocerebellar axonal projection pattern.

Single axonal morphology and termination to cerebellar aldolase C stripes characterize distinct spinocerebellar projection systems originating from the thoracic spinal cord in the mouse.

The spinocerebellar projection has an essential role in sensorimotor coordination of limbs and the trunk. Multiple groups of spinocerebellar projections have been identified in retrograde labeling studies. In this study, we aimed at characterizing projection patterns of these groups using a combination of anterograde labeling of the thoracic spinal cord and aldolase C immunostaining of longitudinal stripes of the cerebellar cortex in the mouse. We reconstructed 22 single spinocerebellar axons, wholly in the cerebellum and brain stem and partly, in the spinal cord. They were classified into three groups, (a) non-crossed axons of Clarke's column neurons (NCC, 8 axons), (b) non-crossed axons of marginal Clarke's column neurons (NMCC, 7 axons), and (c) crossed axons of neurons in the medial ventral horn (CMVH, 7 axons), based on previous retrograde labeling studies. While NCC axons projected mainly to multiple bilateral stripes in vermal lobules II-IV and VIII-IX, and the ipsilateral medial cerebellar nucleus, NMCC axons projected mainly to ipsilateral stripes in paravermal lobules II-V and copula pyramidis, and the anterior interposed nucleus. CMVH axons projected bilaterally to multiple stripes in lobules II-V with a small number of terminals but had abundant collaterals in the spinal cord and medullary reticular nuclei as well as in the vestibular and cerebellar nuclei. The results indicate that, while CMVH axons overlap with propriospinal and spinoreticular projections, NCC and NMCC axons are primarily spinocerebellar axons, which seem to be involved in relatively more proximal and distal sensorimotor controls, respectively.

Projections from the lowest lumbar and sacral-caudal segments to the cerebellar cortex in the rat: An anterograde tracing study

The crossed spinocerebellar tracts originate from neurons in the basolateral part of lamina V, the sacral nuclei of Stilling and the ventrolateral part of the ventral horn of the L6 to caudal segments. The present study examined their projection areas in the cerebellar cortex by using anterograde labeling of mossy fiber terminals with biotinylated dextran in the rat. Labeled terminals were distributed bilaterally in lobules I–V of the anterior lobe. They were most abundant in the apical parts of the lateral vermis and the intermediate region of lobules Ib and IIa, and the rostral side of lobule IIb. The number of labeled terminals in lobules Ib-IIb accounted for 56% and 81%, respectively, of the total 9783 and 7045 labeled terminals. The number of labeled terminals decreased in lobules III to V. In the posterior lobe labeled terminals were distributed exclusively to lobules VIIIa and VIIIb and copula pyramidis. The present study demonstrates that spinocerebellar neurons of the sacral-caudal segments project primarily to the lateral part of lobules I and II, and less densely to lobules III–V and VIII, and copula pyramidis. The projection pattern was essentially similar to that observed in the cat.

Transmission of Predictable Sensory Signals to the Cerebellum via Climbing Fiber Pathways Is Gated during Exploratory Behavior.

Pathways arising from the periphery that target the inferior olive [spino-olivocerebellar pathways (SOCPs)] are a vital source of information to the cerebellum and are modulated (gated) during active movements. This limits their ability to forward signals to climbing fibers in the cerebellar cortex. We tested the hypothesis that the temporal pattern of gating is related to the predictability of a sensory signal. Low-intensity electrical stimulation of the ipsilateral hindlimb in awake rats evoked field potentials in the C1 zone in the copula pyramidis of the cerebellar cortex. Responses had an onset latency of 12.5 ± 0.3 ms and were either short or long duration (8.7 ± 0.1 vs 31.2 ± 0.3 ms, respectively). Both types of response were shown to be mainly climbing fiber in origin and therefore evoked by transmission in hindlimb SOCPs. Changes in response size (area of field, millivolts per millisecond) were used to monitor differences in transmission during rest and three phases of rearing: phase 1, rearing up; phase 2, upright; and phase 3, rearing down. Responses evoked during phase 2 were similar in size to rest but were smaller during phases 1 and 3, i.e., transmission was reduced during active movement when self-generated (predictable) sensory signals from the hindlimbs are likely to occur. To test whether the pattern of gating was related to the predictability of the sensory signal, some animals received the hindlimb stimulation only during phase 2. Over ∼10 d, the responses became progressively smaller in size, consistent with gating-out transmission of predictable sensory signals relayed via SOCPs.SIGNIFICANCE STATEMENT A major route for peripheral information to gain access to the cerebellum is via ascending climbing fiber pathways. During active movements, gating of transmission in these pathways controls when climbing fiber signals can modify cerebellar activity. We investigated this phenomenon in rats during their exploratory behavior of rearing. During rearing up and down, transmission was reduced at a time when self-generated, behaviorally irrelevant (predictable) signals occur. However, during the upright phase of rearing, transmission was increased when behaviorally relevant (unpredictable) signals may occur. When the peripheral stimulation was delivered only during the upright phase, so its occurrence became predictable over time, transmission was reduced. Therefore, the results indicate that the gating is related to the level of predictability of a sensory signal.

Structural basis of cerebellar microcircuits in the rat.

The topography of the cerebellar cortex is described by at least three different maps, with the basic units of each map termed "microzones," "patches," and "bands." These are defined, respectively, by different patterns of climbing fiber input, mossy fiber input, and Purkinje cell (PC) phenotype. Based on embryological development, the "one-map" hypothesis proposes that the basic units of each map align in the adult animal and the aim of the present study was to test this possibility. In barbiturate anesthetized adult rats, nanoinjections of bidirectional tracer (Retrobeads and biotinylated dextran amine) were made into somatotopically identified regions within the hindlimb C1 zone in copula pyramidis. Injection sites were mapped relative to PC bands defined by the molecular marker zebrin II and were correlated with the pattern of retrograde cell labeling within the inferior olive and in the basilar pontine nuclei to determine connectivity of microzones and patches, respectively, and also with the distributions of biotinylated dextran amine-labeled PC terminals in the cerebellar nuclei. Zebrin bands were found to be related to both climbing fiber and mossy fiber inputs and also to cortical representation of different parts of the ipsilateral hindpaw, indicating a precise spatial organization within cerebellar microcircuitry. This precise connectivity extends to PC terminal fields in the cerebellar nuclei and olivonuclear projections. These findings strongly support the one-map hypothesis and suggest that, at the microcircuit level of resolution, the cerebellar cortex has a common plan of spatial organization for major inputs, outputs, and PC phenotype.

Projection patterns of single mossy fiber axons originating from the dorsal column nuclei mapped on the aldolase C compartments in the rat cerebellar cortex

Although cerebellar mossy fibers are the most abundant cerebellar afferents and are deeply involved in cerebellar function, the organization of their projection has remained obscure, particularly in relation to cerebellar compartmentalization. The dorsal column nuclei (DCN) are a major source of cerebellar mossy fibers and possess distinct somatotopic representations of specific somatosensory submodalities. We reconstructed individual dextran-labeled DCN axons completely from serial sections and mapped their terminals on the longitudinal cerebellar compartments that were visualized by aldolase C immunostaining to clarify their projection pattern. Individual axons branched and formed about 100 rosette terminals in the cerebellar cortex, but infrequently projected to the cerebellar nuclei (1 out of 15 axons). Cortical terminals were clustered in multiple areas in the vermis and pars intermedia mostly, but not exclusively, ipsilateral to the origin of the axon. The gracile, cuneate, and external cuneate nuclei (ECuN) mainly projected to the copula pyramidis and lobule V, paramedian and simple lobules, and lobules I-V and VIII-IX, respectively, although there was some overlap. The majority of terminals were located within aldolase C negative or lightly positive compartments. However, terminals of a single axon can be located on aldolase C-negative as well as on aldolase C-positive compartments. In particular, the rostral ECuN, which is responsive to shoulder movements, projected consistently to lobule IX, which were mostly aldolase C-positive. In sum, DCN-cerebellar axons project to multiple compartments with terminals clustered mainly in the conventional spinocerebellar region with a coarse topography, which shows some relationship to the cortical compartments defined by aldolase C.

The periaqueductal grey modulates sensory input to the cerebellum: a role in coping behaviour?

The paths that link the periaqueductal grey (PAG) to hindbrain motor circuits underlying changes in behavioural responsiveness to external stimuli are unknown. A major candidate structure for mediating these effects is the cerebellum. The present experiments test this directly by monitoring changes in size of cerebellar responses evoked by peripheral stimuli following activation of the PAG. In 22 anaesthetized adult Wistar rats, climbing fibre field potentials were recorded from the C1 zone in the paramedian lobule and the copula pyramidis of the cerebellar cortex evoked, respectively, by electrical stimulation of the ipsilateral fore- and hindlimb. An initial and a late response were attributable to activation of Abeta and Adelta peripheral afferents respectively (hindlimb onset latencies 16.9 and 23.8 ms). Chemical stimulation at physiologically-identified sites in the ventrolateral PAG (a region known to be associated with hyporeactive immobility) resulted in a significant reduction in size of both the Abeta and Adelta evoked field potentials (mean reduction relative to control +/- SEM, 59 +/- 7.5 and 66 +/- 11.9% respectively). Responses evoked by electrical stimulation of the dorsal or ventral funiculus of the spinal cord were also reduced by PAG stimulation, suggesting that part of the modulation may occur at supraspinal sites (including at the level of the inferior olive). Overall, the results provide novel evidence of descending control into motor control centres, and provide the basis for future studies into the role of the PAG in regulating motor activity in different behavioural states and in chronic pain.

A novel site of synaptic relay for climbing fibre pathways relaying signals from the motor cortex to the cerebellar cortical C1 zone

The climbing fibre projection from the motor cortex to the cerebellar cortical C1 zone in the posterior lobe of the rat cerebellum was investigated using a combination of physiological, anatomical and neuropharmacological techniques. Electrical stimulation of the ipsilateral fore- or hindimbs or somatotopically corresponding parts of the contralateral motor cortex evoked climbing fibre field potentials at the same cerebellar recording sites. Forelimb-related responses were located in the C1 zone in the paramedian lobule or lobulus simplex and hindlimb-related responses were located in the C1 zone in the copula pyramidis. Microinjections of anterograde axonal tracer (Fluoro-Ruby or Fluoro-Emerald) were made into the fore- or hindlimb parts of the motor cortex where stimulation evoked the largest cerebellar responses. After a survival period of 7-10 days, the neuraxis was examined for anterograde labelling. No terminal labelling was ever found in the inferior olive, but labelled terminals were consistently found in a well-localized site in the dorso-medial medulla, ventral to the gracile nucleus, termed the matrix region. Pharmacological inactivation of the matrix region (2 mm caudal to the obex) selectively reduced transmission in descending (cerebro-olivocerebellar) but not ascending (spino-olivocerebellar) paths targeting fore- or hindlimb-receiving parts of the C1 zone. Transmission in spino-olivocerebellar paths was either unaffected, or in some cases increased. The identification of a novel pre-olivary relay in cerebro-olivocerebellar paths originating from fore- and hindlimb motor cortex has implications for the regulation of transmission in climbing fibre pathways during voluntary movements and motor learning.

Pontine Maps Linking Somatosensory and Cerebellar Cortices Are in Register with Climbing Fiber Somatotopy

The cerebropontocerebellar mossy fiber system is a major CNS sensorimotor pathway. We used a double-retrograde axonal tracing technique (red and green beads) to chart in rats the pontocerebellar projection to different electrophysiologically defined climbing fiber zones in the posterior lobe (face-receiving A2 zone and forelimb- and hindlimb-receiving parts of the C1 zone in the paramedian lobule and copula pyramidis, respectively). Individual cortical injection sites were verified as located in a given zone by mapping the pattern of cell labeling in the inferior olive, whereas labeled cells in the pontine nuclei were mapped using computer-aided three-dimensional reconstruction techniques. A number of topographical differences were found for the pontine projection to the individual zones. Projections to the A2 zone were bilateral, whereas to both parts of the C1 zone, the inputs were mainly contralateral. Furthermore, the A2 (face), C1 (forelimb), and C1 (hindlimb) zone projections were centered in progressively more caudal parts of the pontine nuclei with little or no overlap between them. The areas occupied by cell labeling for each zone corresponded closely to territories in the pontine nuclei shown previously to receive projections from somatotopically equivalent regions of the somatosensory cortex. This precise cerebropontocerebellar topography, defined by climbing fiber somatotopy, is a new principle of organization for linking somatosensory and cerebellar cortices. The convergence of direct and indirect sensory projections is likely to have important implications for cerebellar cortical processing.

The Distribution of Climbing and Mossy Fiber Collateral Branches from the Copula Pyramidis and the Paramedian Lobule: Congruence of Climbing Fiber Cortical Zones and the Pattern of Zebrin Banding within the Rat Cerebellum

Individual cerebellar cortical zones defined by the somatotopy of climbing fiber responses and by their olivo-cortico-nuclear connections located in the paramedian lobule and the copula pyramidis of the rat cerebellum were microinjected with cholera toxin B subunit. Collateral branches of climbing and mossy fibers were mapped and related to the pattern of zebrin-positive and -negative bands of Purkinje cells. Climbing fiber collaterals from the copula distribute to the anterior lobe: from the paramedian lobule mainly to lobulus simplex and rostral crus I. Climbing fibers terminating in particular zones (X, A2, C1, CX, C2, C3, D1, and D2) in the paramedian lobule or the copula collateralize to one or two corresponding zones in lobulus simplex, crus I and II, the paraflocculus, and/or the anterior lobe. These zones can be defined by their relationship to the pattern of zebrin banding. Collaterals from mossy fibers, labeled from the same injection sites in the copula and paramedian lobule, often distribute bilaterally in a symmetrical pattern of multiple but ill-defined longitudinal strips in the anterior lobe and/or lobulus simplex. One or more of these longitudinal aggregates of mossy fiber collaterals was always found subjacent to the strip(s) of labeled climbing fiber collaterals arising from the same locus in the paramedian lobule or the copula. Corticonuclear projections focused on the target nucleus of each zone, although a bilateral plexus of thinner axons, presumably of mossy fiber collateral origin, was sometimes also present in several other regions of the cerebellar nuclei. Overall, these results suggest that climbing fiber zones and zebrin banding reflect a common organizational scheme within the cerebellar cortex.

Multiple zonal projections of the nucleus reticularis tegmenti pontis to the cerebellar cortex of the rat.

Compartmentalization (alternating labelled and unlabelled stripes) of mossy fibre terminals was found in the cerebellar cortex after iontophoretic injections of biotinylated dextran amine into discrete regions of the nucleus reticularis tegmenti pontis (NRTP). The zonal pattern was only observed when volumes of nuclear tissue ranging from 4.5 x 106 to 17.66 x 106 microm3 were impregnated. Up to nine compartments (i.e. up to five stripes separated by four interstripes) were found in crus I and in vermal lobule VI. Up to seven compartments (four stripes and three interstripes) were found in crus II; up to five compartments (three stripes and two interstripes) were identified in the lobulus simplex, the paraflocculus and vermal lobules IV, V and VII; up to three compartments (two stripes and one interstripe) were identified in the paramedian lobule and, finally, up to two compartments (one stripe and one interstripe) were identified in the copula pyramidis, in the flocculus and in vermal lobules II, III, VIII and IX. The projections of the NRTP are arranged according to a divergent/convergent projection pattern. From single injections in the NRTP, projections were traced to a set of cortical stripes widely distributed over the cerebellar cortex. The set of stripes labelled from different regions of the NRTP partially overlapped but complete overlap was never found. This finding revealed that the topographic combination of the projections of the NRTP to the cerebellar cortex is specific for each region of the NRTP. Finally, the projections to single cortical areas were arranged according to a pattern of compartmentalization that is specific for each cortical area, independent of the site of injection in the NRTP and of the number of stripes evident in the cortex.

Topological relationship between corticotropin-releasing factor-immunoreactive cerebellar afferents and tyrosine hydroxylase-immunoreactive purkinje cells in a hereditary ataxic mutant, rolling mouse nagoya

Using immunohistochemistry we examined the distribution of corticotropin-releasing factor-positive cerebellar afferents and the topological relationship between their projections and the distribution of tyrosine hydroxylase-positive Purkinje cells in an ataxic mutant, rolling mouse Nagoya. In the mutants, some climbing fibers were more intensely stained for corticotropin-releasing factor, but their zonal distribution remained the same as in non-ataxic littermates (control mice). These climbing fibers arose from the dorsal accessory nucleus, the ventral lamella of principal nucleus, the dorsomedial cell group, the subnucleus A, the β subnucleus and the ventrolateral protrusion of the inferior olive, since perikarya in these olivary subdivisions were more intensely stained for corticotropin-releasing factor than in controls. Some mossy fiber rosettes in the vermal lobules, the simple lobule, the crus I of ansiform lobule, the copula pyramidis and the flocculus also exhibited corticotropin-releasing factor immunoreactivity and were more densely stained in the mutants than in controls. Double immunostaining for corticotropin-releasing factor and tyrosine hydroxylase in the mutant cerebellum revealed that the distribution of tyrosine hydroxylase-positive Purkinje cells corresponded to terminal fields of corticotropin-releasing factor-positive climbing fibers but not corticotropin-releasing factor-positive mossy fibers. This study indicated an increased corticotropin-releasing factor immunoreactivity in some climbing or mossy fibers in the cerebellum of rolling mouse Nagoya. We also found that the distribution of tyrosine hydroxylase-positive Purkinje cells corresponded to terminal fields of corticotropin-releasing factor-positive climbing fibers in the mutant cerebellum. As the transcription of the tyrosine hydroxylase gene is facilitated by Ca 2+, abnormal tyrosine hydroxylase expression in the mutant Purkinje cells may indicate functional abnormality by alterations in intracellular Ca 2+ concentrations. Therefore, we suggest that an increased level of corticotropin-releasing factor in a specific population of climbing fibers may alter the function of their target Purkinje cells.

Multiple zonal projections of the basilar pontine nuclei to the cerebellar cortex of the rat.

This study revealed a sagittal zonal pattern of projections to the cerebellar cortex after hydraulic or iontophoretic injections of anterograde tracers (tritiated leucine, wheat germ agglutinin-horseradish peroxidase, or biotinylated dextrane amine) in the basilar pontine nuclei of Wistar rats. The zonal pattern of projection was observed only after injections of small size, whereas large injections labeled diffusely wide areas of the cerebellar cortex, masking the zonal projection because the fusion of contiguous stripes. Diverging projections to discrete sets of sagittal stripes in the two sides of the cerebellar cortex arose from single injections. The stripes of fiber terminals were sharply delimited on both sides by areas, interstripes, either virtually void of labeling or with a much lower density of labeling. Thus, the areas of the cerebellar cortex were parceled in sets of sagittal compartments, stripes and interstripes, by the pontine projections. Up to five compartments (three stripes and two interstripes) were observed in the paraflocculus, in the copula pyramidis, and in vermal lobule IX. Up to nine compartments (five stripes and four interstripes) were found in the crus I, the lobulus simplex, the paramedian lobule, and vermal lobules VI-VIII. Up to seven compartments (four stripes and three interstripes) were found in the crus II. Single injections into the basilar pontine nuclei usually labeled symmetric areas of the cerebellar cortex, which, in some cases, showed similar number of stripes. When this was not the case, the stripes were usually more numerous in the contralateral than in the ipsilateral side. All areas of the cerebellar cortex were projected upon, with zonation patterns from different regions of the basilar pontine nuclei. The projections of the basilar pontine nuclei to the cerebellar cortex were arranged according to a fixed pattern specific for each cortical area, independently of the number of stripes labeled within. The mean width of the stripes visualized in the single cortical areas of different rats was similar, despite the different size of the injections. The length of the stripes ranged widely in the various areas of different rats. The data collected in this study are consistent with the idea that all the mossy afferents to the cerebellar cortex are arranged with a zonal pattern.

Target velocity signals in frontal gaze-velocity neurons and severe impairments of smooth gaze tracking following muscimol infusion into the frontal eye fields (FEF) of trained monkeys

The projection from the external cuneate nucleus (ECN) to the cerebellum was studied in the gerbil following the retrograde transport of minute injections (0.05–0.1 μl, 30% solution) or implantations of horseradish peroxidase (HRP) in various folia of the cerebellar cortex and deep nuclei. The morphology of the labelled neurons, as well as their quantitative distribution, was also examined. The projections were in general bilateral but predominantly ipsilateral. A remarkable finding, however, was the predominant contralateral projection from the medial cell column of ECN representing the paw region. Topographically, the nuclear areas receiving the thoracic, shoulder and neck muscle afférents projected bilaterally to vermis lobules I–V and VIII–IX, lobulus simplex, crura Ia and Ib, paraflocculus and flocculus, and contralaterally to crus II and nucleus interpositus; the nuclear regions receiving the forelimb muscle afferents sent fibers bilaterally to vermis lobules V–VIII, paramedian lobule and copula pyramidis, and ipsilaterally to crus II and nucleus interpositus. Based on their somal size, three classes of neurons were distinguished: small, medium and large cells. The small and medium cells constituted 79 and 20% of the population of the labelled cells respectively, whereas the large cells were only occasionally identified. In the cell columns of the caudal ECN representing the forelimb (Lan et al. 1994), most labelled neurons were medium cells which made up 65% of the total labelled medium cells. In contrast, the majority (90%) of the labelled cells in the rostral ECN were of the small-sized variety. The functional significance of the segregation, both in the projection patterns and sizes of somata of the cuneocerebellar neurons is discussed.

Somatotopical organisation within the climbing fibre projection to the paramedian lobule and copula pyramidis of the rat cerebellum

The climbing fibre projection to the paramedian lobule (lobule VII) and the copula pyramidis (lobule VIII) in the posterior lobe of the rat cerebellum was investigated in pentobarbitone-anaesthetised animals. Percutaneous electrical stimulation generated climbing fibre field potentials on the cerebellar surface that indicated a somatotopical organisation into five distinct cortical areas. Each area was identified by the site(s) of peripheral stimulation that evoked the largest response within that area, and from medial to lateral the cortex was subdivided as follows [principal site(s) of stimulation in parentheses with corresponding range of onset latencies]: in the paramedian lobule, area 1 (contralateral face, 17-18 ms), area 2 (ipsilateral forelimb, 10-15 ms) and area 3 (ipsi- and contralateral forelimbs, 16-26 ms and 15-30 ms, respectively); and in the copula pyramidis, area 4 (tail, 17-21 ms) and area 5 (ipsilateral hindlimb, 13-19 ms). In additional retrograde tracer experiments, small volumes of fluorescent-tagged beads were injected into each of the different areas and the location of retrogradely labelled olive cells was mapped. By comparison with other species, the results indicate that in the paramedian lobule area 1 corresponds to zone A2; area 2 corresponds, at least in part, to medial C1; and area 3 corresponds to C2; in addition, farther laterally, a C3 zone may be present. In the copula pyramidis, areas 4 and 5 correspond to subzones of medial C1. Overall, the results support the view that a general principle of cerebellar cortical organisation is a division into parasagittal zones, each characterised by its somatotopically organised climbing fibre input that arises from a specific, rostrocaudally oriented column of cells within the inferior olivary nucleus.

Somatotopical organisation within the climbing fibre projection to the paramedian lobule and copula pyramidis of the rat cerebellum

The climbing fibre projection to the paramedian lobule (lobule VII) and the copula pyramidis (lobule VIII) in the posterior lobe of the rat cerebellum was investigated in pentobarbitone-anaesthetised animals. Percutaneous electrical stimulation generated climbing fibre field potentials on the cerebellar surface that indicated a somatotopical organisation into five distinct cortical areas. Each area was identified by the site(s) of peripheral stimulation that evoked the largest response within that area, and from medial to lateral the cortex was subdivided as follows [principal site(s) of stimulation in parentheses with corresponding range of onset latencies]: in the paramedian lobule, area 1 (contralateral face, 17–18 ms), area 2 (ipsilateral forelimb, 10–15 ms) and area 3 (ipsi- and contralateral forelimbs, 16–26 ms and 15–30 ms, respectively); and in the copula pyramidis, area 4 (tail, 17–21 ms) and area 5 (ipsilateral hindlimb, 13–19 ms). In additional retrograde tracer experiments, small volumes of fluorescent-tagged beads were injected into each of the different areas and the location of retrogradely labelled olive cells was mapped. By comparison with other species, the results indicate that in the paramedian lobule area 1 corresponds to zone A2; area 2 corresponds, at least in part, to medial C1; and area 3 corresponds to C2; in addition, farther laterally, a C3 zone may be present. In the copula pyramidis, areas 4 and 5 correspond to subzones of medial C1. Overall, the results support the view that a general principle of cerebellar cortical organisation is a division into parasagittal zones, each characterised by its somatotopically organised climbing fibre input that arises from a specific, rostrocaudally oriented column of cells within the inferior olivary nucleus. J. Comp. Neurol. 389:249–263, 1997. © 1997 Wiley-Liss, Inc.

Metabotropic glutamate receptors mGluR2 and mGluR5 are expressed in two non-overlapping populations of Golgi cells in the rat cerebellum.

The metabotropic glutamate receptor subtypes mGluR2 and mGluR5, which are thought to be coupled respectively to the inhibitory cyclic adenosine monophosphate (cAMP) cascade and the phosphatidylinositol hydrolysis/Ca2+ cascade, are known to be expressed on Golgi cells in the granular layer of the rat cerebellar cortex. In the present immunohistochemical study with a monoclonal antibody against mGluR2 and a polyclonal antibody for mGluR5, we examined whether or not mGluR2- and mGluR5-like immunoreactivities were both present in single Golgi cells in the rat cerebellar cortex. In double immunofluorescence histochemistry, no Golgi cells showed mGluR2- and mGluR5-like immunoreactivities simultaneously. Of the total number of Golgi cells immunoreactive for mGluR2 or mGluR5, about 90% were mGluR2-like immunoreactive, and about 10% were mGluR5-like immunoreactive. Golgi cells with mGluR2-like immunoreactivity were distributed evenly in the granular layer of all the cerebellar regions, while those with mGluR5-like immunoreactivity were distributed more frequently in the I, II, VII-X lobules of the vermis and the copula pyramidis of the hemisphere than in other cerebellar regions. The results indicate that Golgi cells containing mGluR2 are segregated from those possessing mGluR5. These two populations of Golgi cells, each equipped with a different metabolic glutamate receptor coupled to a different intracellular signal transduction system, may play different roles in the glutamatergic neuronal circuits in the cerebellar cortex.

Cerebellovestibular projection from the posterior lobe cortex in the rabbit: an experimental study with the retrograde HRP method. I. Topographical relationships.

The sources of corticovestibular projections from the cerebellar posterior lobe vermis and hemisphere in the rabbit were investigated by the retrograde horseradish peroxidase (HRP) tracing method. Following iontophoretic injections into various subdivisions of the vestibular nuclear complex (VNC), labelled Purkinje neurones were identified ipsilaterally in all vermal lobules (VI through IX) and some regions of the hemisphere. The results indicate that projection is profuse and directed to all four nuclei of VNC. The labelling suggests some topographical relationships between lobules and especially sublobules of the posterior lobe and individual nuclei of VNC. The major projection originates from lobules VI and IX. A moderate projection derives from lobule VIII whereas that from lobule VII is small. Projections from the hemisphere arise mainly from crus I and crus II of the ansiform lobule. Those from the copula pyramidis, lobule HV and the ventral paraflocculus are sparse. In the present study a topographical relationships were more precisely established than before because the technique revealed VNC afferents from individual sublobules of the cerebellar posterior lobe cortex.