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.     

A re-examination of the cerebellar projections from the gracile, main and external cuneate nuclei in the cat

Cerebellar projections from the dorsal column and external cuneate nuclei in the cat have been studied by means of retrograde axonal transport of horseradish peroxidase. Localized injections covering the entire cerebellar cortex and nuclei show that the gracile nucleus has a weak projection only to the cortex of the anterior lobe, but that there is a conspicuous projection from the main cuneate nucleus to the cerebellum. Most of these fibres reach lobule V and the adjascent parts of lobules IV and VI, and there is also a heavy projection to the paramedian lobule. Some fibres reach lobule IX and possibly also lobules II, III and VIIIB, and nuclear afferents also reach the fastigial and interposite nuclei.Three cerebellar cortical regions are the main targets for the fibres from the external cuneate nucleus, viz. lobule V with the adjacent regions of lobules IV and VI, lobules I and II and lobule IX (the anterior part). Other important afferent regions are the paramedian lobule and the cerebellar nuclei, especially the anterior interposite, and some fibres reach the flocculus.The projections are predominantly ipsilateral.The investigation is the first detailed study of the cerebellar projections from the three nuclei and the findings are discussed in relation to previous experimental observations.     

Studies on the cerebellar projections from the main and external cuneate nuclei in the cat by means of retrograde axonal transport of horseradish peroxidase.

The cerebellar projections from the main and external cuneate nuclei in the cat have been studied by means of retrograde axonal transport of horseradish peroxidase. The main projection from the external cuneate nucleus (ECN) is to the intermediate and, possibly, the small lateral part of lobule V and to the paramedian lobule on the ipsilateral side. The projection from the ECN to the cerebellar regions mentioned is topographically organized. Cells in the caudal part of the ECN send their axons to the caudal parts of lobule V and to the rostral part of the paramedian lobule. Cells in the rostral part of the ECN project to the rostralmost part of lobule V and to the folia in the caudal part of the paramedian lobule. The experimental study also shows that cells in the main cuneate nucleus (MCN) send their axons to the cerebellum. These axons, like those from the ECN, terminate in the intermediate part of lobule V of the anterior lobe and in the paramedian lobule. However, the axons of the cells in the MCN terminate only in the superficial parts of the folia, whereas those from the ECN terminate in the depth of the folia in these two cerebellar areas. The present study also gives evidence that cells in the ventral part of the gracile nucleus send their axons to lobules I and II of the anterior lobe vermis. The observations referred to here are to our knowledge the first anatomical findings demonstrating a projection from the main cuneate and gracile nuclei onto the cerebellar cortex. The observations confirm previous physiological studies.     

Olivocerebellar and cerebelloolivary connections of the oculomotor region of the fastigial nucleus in the macaque monkey

Anatomical connections of the caudal portion of the fastigial nucleus (FN) with the inferior olive (IO) were studied in macaque monkeys with wheat-germ-agglutinin-conjugated horseradish peroxidase (WGA/HRP) and HRP. When injected HRP was confined to a caudal portion of the FN, retrogradely labeled Purkinje cells (P cells) appeared in the oculomotor vermis. We defined the area that receives the projection from vermal lobule VII as the fastigial oculomotor region. The same HRP injection resulted in retrograde labeling of IO neurons in an area of group b (of Bowman and Sladek: J. Comp. Neurol. 152:299-316, '73) of the contralateral medial accessory olive (MAO). This area was designated as the Z-portion because in the coronal section it appears like the letter "Z." Retrogradely labeled IO neurons were also found in the Z-portion when HRP was injected into the oculomotor vermis, indicating that neurons in this portion project to both the fastigial and vermal oculomotor regions. Anterogradely labeled axons from the contralateral fastigial oculomotor region also terminated in the Z-portion. When the effective site included a region anterior to the fastigial oculomotor region, labeled P cells appeared in lobule V and labeled IO neurons appeared in group a. Labeled terminals of fastigial fibers were also found in group a. When the effective site included a region ventral to the oculomotor region, labeled P cells appeared in vermal lobules VIII and IX and labeled IO neurons appeared in caudal parts of a and b, in addition to group c. HRP injection into the posterior interposed nucleus (PIN) resulted in labeling of P cells in the paravermal zone and of IO neurons in the rostral two-thirds of the MAO and the dorsal accessory olive (DAO). The location of the labeled terminals coincided with the region where the densest labeling of IO neurons was found. Thus, the olivary projections to both the cerebellar cortex and deep cerebellar nuclei and the nucleoolivary projection exhibited a closely related topographical organization.     

Striatal afferent connections in the turtle (Chrysemys picta) as revealed by retrograde axonal transport of horseradish peroxidase.

Horseradish peroxidase (HRP, 30% solution, 0.1-0.3 mul, 72 h) was injected unilaterally into the basal striatum (STR) and the dorsal ventricular ridge (DVR) of adult turtles (Chrysemys picta) in order to demonstrate the cells of origin of some afferents to these telencephalic structures. After selective STR injection, HRP-labeled cells were visualized in the dorsal thalamus and midbrain tegmentum, ipsilaterally. At thalamic level, HRP-positive neurons were located around nucleus rotundus, i.e., mainly within nuclei dorsomedialis anterior, dorsolateralis anterior and less abundantly in nuclei ventralis and reuniens. At midbrain level, a large population of labeled neurons was disclosed within the ventrolateral portion of rostral tegmentum. Other HRP-positive neuronal somata were found scattered throughout the lateral portion of the caudal midbrain tegmentum. In addition, labeled axons were visualized in both peduncles of the lateral forebrain bundle (LFB) after STR injection. The HRP-positive fibers of the dorsal peduncle of the LFB were followed up to the ipsilateral labeled thalamic cells where they appear to arise, whereas the HRP-containing axons of the ventral peduncle were traced down to the lateral midbrain tegmentum where they appear to arborize. Most of the HRP injections into the DVR were confined to the mediodorsal quadrant of the rostral half of the DVR. In such a case, a very large number of HRP-positive cells were disclosed within all thalamic nuclei surrounding nucleus rotundus, ipsilaterally. In addition, numerous labeled neurons were also found in nucleus rotundus itself and within nucleus reuniens. No HRP-positive cells were disclosed caudally to the meso-diencephalic junction after DVR injection.     

Secondary trigeminocerebellar projections in sheep studied with the horseradish peroxidase tracing method

Secondary trigeminocerebellar connections have been studied with HRP histochemistry in 25 sheep. The results indicate that almost all of the cerebellar cortex except flocculus, ventral paraflocculus and lobules I-IV receives bilateral (mostly ipsilateral) fibers from the trigeminal nuclei. A topographical organization of trigeminocerebellar fibers is present. The mesencephalic tract nucleus projects to the anterior lobe, the simple lobule (HVI), lobules VI, VIII, and the dorsal paraflocculus. The ventral group of the princeps and spinal tract (mainly IDV) nuclei projects to all lobules studied in vermis and hemispheres. More dorsal parts of these nuclei have a more restricted projection field including the vermal lobules VI, VII, and IX and the hemisphere. Cells within and ventral to the motor nucleus of the trigeminal nerve were found labeled after injections into the anterior lobe, the simple lobule, and lobule IX. Labeled cells in the region of the nucleus ovalis and close to the solitary tract project to the simple and paramedian lobule and lobule IX.     

Brain stem afferents to the fastigial nucleus in the cat demonstrated by transport of horseradish peroxidase.

Although retrograde and anterograde degeneration studies have provided important information concerning brain stem afferents to the fastigal nucleus (FN), these data may be incomplete and should be confirmed by axonal transport methods. Attempts were made to inject horseradish peroxidase (HRP) unilaterally into the FN in a series of adult cats. Animals were perfused with dextran and a fixative solution of paraformaldehyde and glutaraldehyde in 0.1 M phospate buffer. Representative sections were treated by the Graham and Karnovsky ('66) method. Selective HRP injections in one FN resulted in retrograde transport of the marker to Purkinje cells of the ipsilateral vermis and distinctive appendages of the contralateral medial accessory olivary (MAO) nucleus (nucleus beta and the dorso-medial cell column). Retrograde transport of the label was found bilaterally in cells of the medial (MVN) and inferior (IVN) vestibular nuclei, in cell group x and in the nucleus prepositus (PP). Labeled vestibular neurons, most numerous in MVN, were identified in dorsal, caudal and lateral regions, with a slight ipsilateral preponderance. Only a few neurons in caudal, dorsal and lateral regions of the IVN were labeled and none of these included cells of group f. Labeled cells in the caudal third of PP were greatest ipsilaterally. Rostral and caudal injections of FN labeled smaller numbers of cells in MVN, IVN, cell group x and PP. HRP injections of FN and portions of lobules VIII and IX resulted in bilateral retrograde labeling of larger numbers of cells in MVN, IVN and cell group x, and ipsilateral labeling of cells in group y and the interstitial nucleus of the vestibular nerve. Injections of HRP into basal folia of lobules V and VI resulted in retrograde transport of the marker to cells of the medial and dorsal accessory olivary nuclei contralaterally, and to cells of the ipsilateral accessory cuneate nucleus. Transport of label injected into portions of the pyramis was detected in parts of the contralateral MAO and bilaterally in parts of the pontine and reticulotegmental nuclei. This study suggests that the principal afferents of the fastigial nucleus arise from: (1) Purkinje cells of the ipsilateral vermis, (2) restricted portions of the contralateral MAO (nucleus beta and dorsomedial cell column), (3) portions of the MVN and IVN (bilaterally) and (4) caudal parts of the PP. Secondary vestibular inputs to the fastigial nucleus probably are relayed mainly by Purkinje cells in the cerebellar cortex.     

Organization of projections from the gracile, medial cuneate and lateral nuclei in the North American opossum. Horseradish peroxidase study of the cells projecting to the cerebellum, thalamus and spinal cord

Using the horseradish peroxidase technique on the North American opossum, we were able to locate the neurons within the dorsal column and lateral cuneate nuclei which innervate the cerebellum and thalamus as well as those within the dorsal column nuclei which project spinalward. The medial and lateral cuneate nuclei supply axons to the anterior lobe, the paramedian lobule and the pyramis of the cerebellum and the lateral nucleus provides an additional projection to the uvula. The cerebellar projections from these nuclei arise from neurons located rostral to the obex. The thalamic projections from the gracile and medial cuneate nuclei originate from neurons throughout their rostral to caudal extent, although most of them are located just rostral to the obex. Neurons within the lateral cuneate nucleus which innervate the thalamus are found at intermediate rostrocaudal levels where most of them approximate the medial cuneate nucleus. The medial cuneate also projects to at least lumbar levels of the spinal cord in the opossum and neurons giving rise to such connections are found at the level of the obex and caudal to it. Neurons within the dorsal part of the dorsal column nuclei were labelled only after thalamic injections. Our results in the opossum are compared with those obtained in several placental mammals.     

Topographical organization in the origin of serotoninergic projections to different regions of the cat cerebellar cortex

The distribution of serotonin immunoreactivity in the cat cerebellum was studied by using the indirect antibody peroxidase-antiperoxidase (PAP) technique. Furthermore, the origin of these chemically defined afferents was determined by combining the retrograde transport of horseradish peroxidase (HRP) with the PAP technique. In the cerebellar cortex, serotonin immunoreactivity is present in a plexus of beaded fibers that is confined almost exclusively to the granule and Purkinje cell layers; a few fibers are present in the molecular layer. Serotoninergic axons and varicosities have a dense and uniform distribution throughout all lobules of the cerebellum with the exception of lobule X where the fiber density is sparse. Serotonin cell bodies were not found within the cerebellar cortex. However, following pretreatment with pargyline and L-tryptophan, serotonin positive cell bodies were found in all deep cerebellar nuclei as well as the raphe and reticular nuclei in the brainstem. The present study demonstrates that the serotoninergic projection to the cat's cerebellum has some degree of topographical organization. Serotoninergic fibers in the anterior vermis (lobules I-V) were shown to arise from neurons located within the paramedian reticular nucleus, the lateral reticular nucleus, and the lateral tegmental field. Injections of HRP into either the posterior vermis (lobule VI-IX) or the paramedian lobule, labeled serotoninergic neurons exclusively in the lateral reticular nucleus. Lobus simplex, crus I and crus II (the hemisphere) receive a serotoninergic input from cells located in the lateral tegmental field, the peri-olivary reticular formation and the paramedian reticular nucleus. In no cases were neurons in the raphe double-labeled, although there were cells positive for HRP or serotonin alone. The data indicate that there is a topographical organization in the serotoninergic projection from the caudal brainstem to specific regions of the cat's cerebellar cortex. In addition to climbing and mossy fibers, this projection represents a third major source of cerebellar afferents based on its dense and widespread distribution as well as its morphological and chemical characteristics.     

Postsynaptic dorsal column pathway of the rat. III. Distribution of ascending afferent fibers

The distribution in the dorsal column nuclei (DCn) of post-synaptic dorsal column (PSDC) fibers was examined in rats following injections of Phaseolus vulgaris leucoagglutinin (PHA-L) in the spinal cord. Lemniscal neurons in the DCn were retrogradely labeled in the same animals by injecting the thalamus with Fluoro-Gold. In some experiments, primary afferent fibers were also labeled by injecting dorsal root ganglia with choleragenoid-conjugated HRP. Injections of PHA-L into the cervical enlargement labeled many fibers and varicosities throughout most of the ipsilateral cuneate nucleus. Labeled fibers were also present in the external cuneate and internal basilar nuclei. Injections of PHA-L into thoracic cord labeled fibers and varicosities in the medial cuneate and lateral gracile nuclei, as well as the external cuneate nucleus. Injections into the lumbar enlargement labeled fibers and varicosities throughout most of the gracile nucleus. Injections in sacral cord labeled fibers in the most medial part of the gracile nucleus. Dense labeling of PSDC fibers was found in areas with high densities of retrogradely labeled lemniscal neurons and areas with high densities of primary afferent fibers. In all regions of the DCn and in the external cuneate nucleus, fibers and varicosities labeled for PHA-L were seen in apposition to retrogradely labeled lemniscal cells. The distribution of postsynaptic afferent fibers in the DCn of the rat and its relationship to lemniscal neurons and primary afferent fibers contrast sharply with these features in cats.     

Origins of cerebellar mossy and climbing fibers immunoreactive for corticotropin-releasing factor in the rabbit

Corticotropin-releasing factor (CRF) has been implicated by both anatomical and physiological techniques as a potential cerebellar transmitter or modulator. In the present experiment, with the aid of immunohistochemistry, we have described specific cerebellar afferent pathways in the rabbit in which CRF is located. CRF-immunoreactive climbing fibers were present in the molecular layer throughout the cerebellum, but especially in lobules 8–9a. All inferior olivary neurons were CRF-immunoreactive. In lobules 8–9a, CRF-immunoreactive mossy fibers were organized in sagittal bands. The highest density of CRF-immunoreactive mossy fiber terminals was observed in the granule cell layer of lobules 8–9a and the flocculus. No CRF-immunoreactive perikarya were located in rabbit cerebellum. The brainstem origin of CRF-immunoreactive mossy fiber terminals was suggested by numerous CRF-immunoreactive perikarya located in the medial, lateral and descending vestibular nuclei, nucleus prepositus hypoglossi, nucleus x, paramedian reticular nucleus, gigantocellular reticular nucleus, lateral reticular nucleus, and raphe nuclei. Using double label experiments, we investigated the specific CRF afferent projection to the flocculus and posterior vermis. Horseradish peroxidase (HRP) injections into the posterior vermis double labeled CRF-immunoreactive neurons in the caudal medial and descending vestibular nuclei and nucleus prepositus hypoglossi. HRP injections into the flocculus double labeled more CRF-immunoreactive neurons in the nucleus prepositus hypoglossi than in the vestibular nuclei. HRP injections into either the posterior vermis or flocculus double labeled CRF-immunoreactive neurons in the paramedian reticular nucleus, nucleus reticularis gigantocellularis, and raphe nuclei. These data suggest that CRF may play an important role in vestibularly related functions of the cerebellum. © 1993 Wiley-Liss, Inc.     

Projection systems and terminal localization of dorsal column afferents: an autoradiographic and horseradish peroxidase study in the rat

Projection systems from the gracile nucleus and the cuneate nuclear complex to their terminal sites in the mesencephalon, diencephalon, and cerebellum were examined by means of anterograde autoradiography and retrograde horseradish peroxidase methods. Three projection systems emerge from the dorsal column nuclei, decussate via internal arcuate fibers, and form the contralateral medial lemniscus (ML). At the obex, some fibers split off the ML and course dorsolaterally, forming an ascending lateral system which fits the "lemniscal adjunct channel" (LAC) concept of Graybiel ('72). The ML continues rostrally as the "main lemniscal line channel" (MLLC). At the inferior colliculus, some LAC fibers terminate in the pontine nuclei, parabrachial, dorsal reticular nuclei, and the external and ventral medial part of the central nucleus of the inferior colliculus. More rostrally at the level of the superior colliculus, terminal fields are found in the medial nucleus of the medial geniculate body, the suprageniculate, pretectal, and mesencephalic reticular nuclei, marking the end of the LAC. In the diencephalon, gracile fibers leave the MLLC and form a crescentlike terminal field along the extreme lateral border of the ventral posterior lateral nucleus (VPL) of the thalamus. Cuneate MLLC fibers terminate in a bandlike formation in the VPL medial to the gracile termination. The third fiber system, the cuneocerebellar projection, emerges from the cuneate, the external cuneate nuclei, and the "cellular bridge" and immediately enters the ipsilateral inferior cerebellar peduncle. Upon entering the cerebellum, the major fiber component remains ipsilateral and terminates as vertical bands in vermal and paravermal lobules, and lobules I through IVa. The posterior cerebellar lobe contains terminal bands in lobules VII-IX, the copula pyramidis, and the paramedian lobule. It is concluded that the dorsolateral fiber system conforms to Graybiel's LAC. It is more divergent and probably less modality specific, whereas the medial lemniscal system conforms to the MLLC, which is said to be modality specific, less divergent, and locked to specific sensory-motor response characteristics. The topography of cerebellar terminal bands indicates that there is sensory-motor representation from all parts of the body to all parts of the cerebellum, at least in the rat.     

Trigeminal and dorsal column nuclei projections to the anterior pretectal nucleus in the rat

The projections of the trigeminal (V) sensory nuclei (VSN) and the dorsal column nuclei (DCN) to the anterior pretectal nucleus (APT) of the rat were investigated by the use of anterograde and retrograde transport of wheat-germ agglutinin-conjugated horseradish peroxidase (WGA-HRP). Injections of WGA-HRP into the APT retrogradely labeled neurons in the contralateral VSN and DCN. The labeled neurons in the VSN were most concentrated in the rostral V subnucleus interpolaris (Vi), but were also found in caudal V subnucleus oralis (Vo). No labeled neurons were seen in V subnucleus caudalis. In the DCN, retrogradely labeled neurons were observed in rostral portions of both the cuneate (Cu) and gracile (Gr) nuclei. Injections of WGA-HRP into the rostral Vi or caudal Vo resulted in dense anterograde terminal labeling in the ventral two-thirds of the APT; the labeling was maximal in the ventromedial part of the caudal half of the APT and did not extend into its most rostral portion. Labeling resulting from injections of tracer into Cu or Gr was located primarily in the ventral half of the APT, was maximal in the mid-levels of the nucleus and extended into its rostral portions. These results indicate the existence of prominent somatosensory projections to the APT and are consistent with recent findings suggesting a role for the APT in sensorimotor integration.     

The overlap of spinothalamic and dorsal column nuclei projections in the ventrobasal complex of the rat thalamus: a double anterograde labeling study using light microscopy analysis

Projections from the spinal cord and the dorsal column nuclei (DCN) to the ventrobasal complex of the thalamus (VB) were studied in the rat by using double anterograde labeling strategy. This strategy was based on the injection of 3H-leucine into the DCN and of wheat germ agglutinin conjugated to horseradish peroxidase (WGA-HRP) into the spinal cord and their subsequent transport. Adjacent 30-micron-thick sections were then processed differentially for autoradiography or for HRP by using tetramethyl benzidine (TMB) as a chromogen. Similar areas of the ventrobasal complex were labeled, in adjacent sections, after a large injection of 3H-leucine into the DCN and when wheat germ agglutinin-HRP had been injected in any part of the spinal cord. If, however, a small injection of the radioactive tracer was centered in the gracile nucleus and compared with an injection of WGA-HRP placed in the lumbar enlargement of the cord, the rostral and dorsal portions of the lateral VB were labeled from both sources. On the other hand, if tritiated leucine was injected into the cuneate nucleus, and WGA-HRP placed in the cervical enlargement, then the caudal and ventral portions of the lateral VB demonstrated overlap of both labels. The present results show that, in the rat, areas of termination of both the spinothalamic tract and the lemniscal pathway originating from the DCN overlap in the lateral VB. This overlap is somatotopically organized, thus indicating that the same area of the VB receives somatic inputs from one particular part of the body through both pathways. These results are discussed in comparison to those of comparable studies performed in the cat and in the monkey and with reference to the electrophysiological data that have demonstrated that, in the rat VB, neurons responding to noxious stimulation are intermingled with neurons exclusively responding to non-noxious stimulation.     

Parasagittal organization of mossy fiber collaterals in the cerebellum of the mouse

We have observed that WGA-HRP injections in lobule VIII of the mouse result in the labeling of mossy fiber terminals in the anterior lobe (lobules I-V), which are distributed in five distinct parasagittal bands. Injections in the anterior lobe label mossy fiber terminations in lobules VIII and IX. We interpret these results as indicating that an extensive system of mossy fiber collaterals exists between the anterior lobe and lobule VIII (less so to IX), which terminates as discrete parasagittal bands in the anterior lobe. Intermediate bands are thus occupied by fibers that do not send collaterals to the posterior vermis (VII-IX). In an attempt to identify the source(s) of this collateral system we have used double retrograde tracing techniques. Following injections of one tracer in the anterior lobe and another in lobule VIII we observe large numbers of double retrogradely labeled neurons in the lateral reticular nucleus, the basilar pontine nuclei, and the spinal cord. Thus, these mossy fiber sources are the most likely origins for the banded collateral system. Our studies do not allow us to distinguish whether one, or more than one, of these regions contribute to the system.     

Organization of spinal inputs to the perihypoglossal complex in the cat

First- and second-order spinal afferents to the perihypoglossal complex were sought by using axonal transport of WGA-HRP. Injections in C1, 2, and 3 dorsal root ganglia resulted in axonal labeling in the nucleus intercalatus and the external cuneate nucleus, with a number of retrogradely labeled cells seen as well in the latter. A similar pattern of axonal labeling in the nucleus intercalatus as well as several retrogradely labeled cells were found after spinal cord injections at levels C1, 2, and 3. A prominent field of labeled axons was also present in the rostral main cuneate nucleus. No labeling was seen in the perihypoglossal nuclei after injections in the spinal cord or dorsal root ganglia at levels caudal to C3. After injections of HRP into the perihypoglossal nucleus we were able to identify labeled neurons within Rexed's laminae V-VIII and the central cervical nucleus. Anterograde labeling in the main cuneate nucleus was observed after C1 to C5 ganglion and C1 to C6 cord injections. The pattern and extent of labeling in the perihypoglossal nuclei and adjacent structures seen after cerebellar injections into lobules V and VI were comparable to those previously reported and permitted evaluation of the relay from dorsal root ganglia through the intercalatus to the vermis. Topography of the cervical projections to the nucleus intercalatus is considered with respect to that of the perihypoglossal-collicular projection. A discussion is offered of the apparent importance of nucleus intercalatus as a relay of cervical and vestibular afferent information to premotor structures involved in neck motor control. The perihypoglossal complex is viewed as being organized in such a fashion as to allow the nuclei intercalatus and prepositus hypoglossi to function as key structures in the integration of inputs related to neck and ocular motor control, respectively.     

Projections to the inferior olive of the cat. II. Comparisons of input from the gracile, cuneate and the spinal trigeminal nuclei

The present experiments compared the projections to the inferior olive of the cat from the gracile, cuneate and spinal trigeminal nuclei. A differential labeling strategy was used for these comparisons. It was found that all three somatic sensory nuclei project to portions of all three major divisions of the contralateral inferior olive. The spinal trigeminal n. may also project less densely to the ipsilateral medial accessory olive. Projections to the dorsal accessory nucleus (DAO) and the medially-adjacent ventral lamella of the principal nucleus are roughly somatotopically organized. Although there is considerable overlap between the projection zones, the gracile n. projects predominantly to lateral DAO, the cuneate n. projects predominantly to medial DAO, and the spinal trigeminal nucleus pars caudalis projects predominantly to the most medial portions of DAO and the ventral lamella of principal olive. Projections to the medial accessory olive, on the other hand, are not as highly organized. Instead, they overlap extensively within a small egg-shaped area in the middle of the caudal half of the nucleus. Whereas all portions of the gracile and cuneate nuclei project to the inferior olive, only the pars caudalis of the spinal trigeminal nucleus appears to do so. These results were compared with the three available olivocerebellar maps as well as with the available behavioral and electrophysiological evidence on cerebellar somatotopic organization. This comparison indicated that the inputs to the cerebellum from the three second-order somatosensory nuclei via the inferior olive appear to be generally consistent with cerebellar somatotopic organization. This consistency is apparent not only with respect to the longitudinally-organized, vermal and paravermal differences in the anterior lobe, but also with respect to the transversely-organized specific somatotopy of the intermediate zone of the anterior lobe and the paramedian lobule.     

Central projections of the utricular nerve in the gerbil

The central projections of primary afferent fibers in the utricular nerve, which convey linear head acceleration signals to neurons in the brainstem and cerebellum, are not completely defined. The purpose of this investigation was twofold: 1) to define the central projections of the gerbil utricular afferents by injecting horseradish peroxidase (HRP) and biotinylated dextran amine (BDA) into the utricular macula; and 2) to investigate the projections of individual utricular afferents by injecting HRP intracellularly into functionally identified utricular neurons. We found that utricular afferents in the gerbil projected to all divisions of the vestibular nuclear complex, except the dorsal lateral vestibular nucleus. In addition, terminals were observed in the interstitial nucleus of the eighth nerve, nucleus Y, external cuneate nucleus, and lobules I, IV, V, IX, and X of the cerebellar vermis. No projections appeared in the flocculus or paraflocculus. Fibers traversed the medial and intermediate cerebellar nuclei, but terminals appeared only occasionally. Individual utricular afferents collateralize extensively, projecting to much of the brainstem area innervated by the whole of the utricular nerve. This study did not produce complete filling of individual afferent collateral projections into the cerebellar cortex.     

Afferent and efferent connections of the oculomotor cerebellar vermis in the macaque monkey

Saccadic eye movements were evoked with weak currents applied to a circumscribed vermal area. The area was confined to lobule VII in the majority of the monkeys and coincided with the distribution of saccade-related neural activity. We defined this area as the oculomotor vermis and studied its anatomical connections with wheat germ-agglutinin conjugated horseradish peroxidase (WGA/HRP) and HRP. When injected HRP was confined to the oculomotor vermis, most labeled Purkinje axons terminated ipsilaterally in an ellipsoidal region in the mediocaudal aspect of the fastigial nucleus. Retrogradely labeled cells were found in two relatively circumscribed regions in the fastigial nucleus: one group was in the lateral half of the ellipsoidal terminal region and the other group was in a spherical region near the lateral margin of the nucleus. Following the injection of HRP into the oculomotor vermis, the largest population of retrogradely labeled neurons was found in the nucleus reticularis tegmenti pontis. Labeled cells were located only in the medial and dorsolateral portions of the nucleus. The cell aggregates in the dorsolateral portion merged with densely labeled cells of the processus tegmentosus lateralis. The second largest population of labeled cells was found in the pontine nuclei. Approximately 28% of the labeled pontine cells aggregated in the paramedian pontine nucleus, whereas the other labeled pontine cells were widely distributed in the dorsal part of the pontine peduncular nucleus and the dorsolateral pontine nucleus. Labeled cells were scattered also in the pontine raphe, the paramedian pontine reticular formation, and the interfascicular nucleus at the rostral level of the hypoglossal nucleus. Fewer labeled cells were discovered in the vestibular nuclear complex and the prepositus hypoglossi. In the inferior olivary nucleus, labeled cells were located in the subnucleus b of the medial accessory nucleus.     

Calcitonin gene-related peptide in afferents to the cat's cerebellar cortex: distribution and origin.

In the present study, the distribution and origin of calcitonin gene-related peptide (CGRP) were analyzed in the cat's cerebellum. Following incubation in an antibody generated against rat CGRP and processing with the peroxidase anti-peroxidase (PAP) technique, CGRP immunoreactivity (IR) is found in profiles that have morphological characteristics of both simple and complex mossy fibers. However, all mossy fibers are not CGRP-positive. Further, CGRP-IR mossy fibers have a heterogeneous distribution in the cerebellum. In the vermis, the majority of immunoreactive profiles are in lobules VII, VIII, and the dorsal folia of IX. In anterior vermal lobules, only scattered terminals, located primarily at the apex and along the shoulder of the folia, are present. Laterally, CGRP-IR mossy fibers are located in the paramedian lobule, paraflocculus, and crus II. No CGRP fibers or varicosities are observed in any of the cerebellar nuclei. However, CGRP-positive cell bodies are scattered throughout the nuclear neuropil. A double label technique revealed that CGRP-IR mossy fibers arise from neurons located in the lateral reticular nucleus, external cuneate nucleus, inferior vestibular nucleus, and basilar pons. The present findings, taken together with previous data, indicate that cerebellar afferents are chemically heterogeneous. The findings of the present study suggest that precerebellar nuclei that give rise to the mossy fibers that contain CGRP have the potential for playing a complex role in modulating circuitry in the cerebellar cortex of the cat.