Medial Accessory Olive Research Articles

Columnar organisation of the inferior olive projection to the posterior lobe of the rat cerebellum

The organisation of the olivocerebellar projection to lobules VI, VIII, and IX of the posterior lobe of the rat cerebellum was investigated in detail by using the retrograde tracer wheat germ agglutinin-horseradish peroxidase. Small, well-defined rostro-caudally orientated columns of olive cells were found to project to different parasagittal areas in the posterior lobe. A column of olive cells about 2,000 microns in rostro-caudal length in subnucleus "c" and nucleus beta of the caudal medial accessory olive (MAO) provides climbing fibre input to the most medial part of lobules VI and IX, but this projection is displaced laterally in lobule VIII by a projection from a column of cells about 600 microns in rostro-caudal length in lateral caudal MAO (subnucleus "a"). It is possible that each of these columns of olivary neurones may be further subdivided in the rostro-caudal axis so that different sections project to different medio-lateral parts of the cortex. A fine-grain 'sublobular' localisation may also exist: the projection to midline lobule VIc arises at caudal levels of the olive from a band of cells in the transition region between subnucleus "c" and nucleus beta, whilst by comparison the projection from caudal levels of the olive to lobules VIa and VIb arises from cells located more ventrally in nucleus beta. Evidence is also presented to confirm that the posterior lobe vermis in the rat extends further laterally than in other mammals and that part of it receives a projection from a column of olive cells, 1,000 microns in rostro-caudal length, in a newly defined region of caudal MAO, termed subnucleus "b1."

Intracellular labeling of neurons in the medial accessory olive of the cat: I. Physiology and light microscopy

This study is the first of three reports on the detailed morphology of horseradish peroxidase injected neurons in the medial accessory olive of the cat. Intracellular, in vivo recordings of olivary cells were made and their response to mesodiencephalic stimulation was tested. In 44 units a short latency action potential could be recorded, which was very suggestive of a monosynaptic excitatory pathway. The short latency response was frequently followed by a long latency (mean 188 msec) or rebound action potential. Recordings were followed by intracellular iontophoresis of horseradish peroxidase. A total of 21 neurons, all located within the medial accessory olive were chosen for morphological analysis. Cells could be divided into two categories on the basis of their overall morphological appearance. Type I cells (n = 5) had sparsely branching dendrites that radiated away from the soma and were usually found in the caudal part of the medial accessory olive. The axon usually originated from the soma. Type II cells (n = 16) were located more rostrally. They had larger cell bodies with dendrites that ramified extensively, forming a globular structure (mean diam. 338 microns). The axon usually originated from a first order dendrite. No recurrent axon collaterals were observed on either type I or II cells. Both cell types carried long and complex spiny appendages; however, they were most numerous on the second and higher order dendrites of type II cells. Since the soma of these cells is usually not found in the centre of its dendritic field, even if the cell is located in the center area of the neuropil, it is suggested that the dendritic trees of up to 100 neurons may be intricately interwoven, establishing clusters with intensive intercommunication by means of dendritic gap junctions. The abundance, length and complexity of the spiny appendages suggest an important role in this process, but may also be relevant instruments in enhancing the computational capabilities of these neurons, especially in time sensitive processes. When relating the physiological and the morphological results, it was noted that both type I and type II cells could respond to mesodiencephalic stimulation and were both able to trigger a rebound action potential. No significant correlations were found between cell size and the latency of the rebound.

Intracellular labeling of neurons in the medial accessory olive of the cat: II Ultrastructure of dendritic spines and their gabaergic innervation

In order to describe the morphology of dendritic spines of identified neurons in the cat inferior olive together with their gamma-aminobutyric acid (GABA) synaptic input, a technique was used combining intracellular labeling of horseradish peroxidase with postembedding gold-immunocytochemistry. With this technique physiologically identified olivary cells were reconstructed with the light microscope, and the horseradish peroxidase reaction product and immunogold labeling were subsequently examined in serial sections at the ultrastructural level. In addition, a degenerating neuron was observed, resulting in a triple labeling in single ultrathin sections. Quantitative and three-dimensional analysis showed that the dendritic spines were composed of long, thin stalks ending in one or more spine heads. The spines of cells located in the caudal half of the medial accessory olive (type I cells, characterized by dendrites which run away from the soma) were found to be less complex than those of cells located rostrally in this olivary subnucleus (type II cells, characterized by dendrites which tend to turn back towards the soma). Most, if not all, of the spines of both cell types were located within glomeruli. On average, the spines within individual glomeruli originated from 6 different dendrites (with a maximum of 8). Different spines within the same glomerulus were never derived from different dendrites of the same olivary neuron, but single spines frequently gave rise to several spine heads, which could be located either within different glomeruli or inside a single glomerulus. The glomerular spine heads originating from the same spine were rarely located near one another. All spines and most of the spine heads were contacted by both GABAergic and non-GABAergic terminals. Most of the GABAergic terminals contained pleomorphical vesicles and displayed symmetric synapses whereas the non-GABAergic terminals showed usually round to oval vesicles and asymmetric synapses.

Intracellular labeling of neurons in the medial accessory olive of the cat: III. Ultrastructure of axon hillock and initial segment and their GABAergic innervation

The gamma-aminobutyric acid (GABA) synaptic input of identified axons in the cat inferior olive was studied by use of combination of intracellular labeling with horseradish peroxidase and postembedding gold-immunocytochemistry. With this technique olivary cells were physiologically identified and light microscopically reconstructed, and the horseradish peroxidase reaction product and the immunogold labeling were subsequently simultaneously visualized for electron microscopic investigation with the use of serial ultrathin sections. The axons of cell type I (characterized by dendrites which radiate away from the cell body) originated from the soma, whereas those of type II neurons (characterized by dendritic trees which curve back towards the soma) were derived from a primary dendrite. The axons of olivary neurons stand out by the length of their axon hillock (up to 21 microns) and initial segment (up to 40 microns). The hillock forms various spiny appendages which were located within glomeruli together with dendritic spines of other olivary neurons. Axonal spines of type II neurons were more numerous and complex looking than those of type I. The axonal spines, the shaft of the axon hillock, and the transition between the hillock and initial segment were primarily innervated by GABAergic terminals (65%) but non-GABAergic terminals (35%) were present as well. The terminals apposed to the axons of type I neurons contacted mainly the axonal shafts, whereas most of the terminals adjacent to the axons of type II neurons established synaptic contacts with the axonal spines. The initial segments were largely devoid of synaptic input. Distally, the initial segment acquired a myelin sheath.

Afferents to the cerebellar flocculus in cat with special reference to pathways conveying vestibular, visual (optokinetic) and oculomotor signals

Horseradish peroxidase (HRP) was injected into the cerebellar flocculus of 20 cats to determine: (a) the proportions of afferents from the various brain stem nuclei; (b) possible projections from the basilar pontine nuclei; and (c) sources of saccadic eye movement signals recorded from flocculus Purkinje cells. Results confirm earlier findings that the flocculus receives large numbers of mossy fibre afferents from the vestibular and perihypoglossal nuclei, bilaterally, and climbing fibres from the contralateral inferior olive (dorsal cap, ventrolateral outgrowth, medial accessory olive, ventral bend of principal olive). In addition, large numbers of HRP-labeled neurons have been identified within: (i) the basilar pontine nuclei, bilaterally, where they are distributed in columns in the dorsolateral, lateral, ventral medial and dorsomedial nuclei; (ii) the nucleus reticularis tegmenti pontis; (iii) several of the cranial motor nuclei, VI, VII, X (retrofacial n.), XI (n. ambiguus), and XII; (iv) the raphe magnus, pontis and obscurus; (v) the lateral reticular nucleus, pars subtrigeminalis. Finally, new information is presented which shows that large numbers of flocculus projecting neurons are located within the medial longitudinal fasciculus at two locations; one just rostral to the hypoglossal nucleus and another group extends 2-3 mm rostral to the abducens nucleus. These groups are bilateral, and have been termed, respectively, the caudal and intermediate interstitial nucleus of the medial longitudinal fasciculus. Both groups correspond in location to physiologically identified neurons in cat which fire in relation to saccadic eye movements. Their projection to the flocculus, in part, explains the saccadic discharge of Purkinje cells in the flocculus.

Cerebellar nucleo‐olivary projections in the rat: An anterograde tracing study with Phaseolus vulgaris‐leucoagglutinin (PHA‐L)

In order to evaluate the reciprocity of olivo-cerebellar and cerebello-olivary connections, a detailed description of the cerebellar nucleo-olivary projection in the rat is presented using small, iontophoretic injections of the anterograde tracer Phaseolus vulgaris-leucoagglutinin. Sparse projections were found to arise from the rostral part of the medial cerebellar nucleus toward the lateral part of the caudal medial accessory olive. Its medial parts receive a projection from the dorsolateral protuberance of the medial cerebellar nucleus. Caudal and lateral regions of the medial cerebellar nucleus project to the "beta" group and dorsomedial cell column. Heavy olivary projections to circumscribed parts of the inferior olive were found after injections in the remaining cerebellar nuclei. The medial part of the posterior interposed nucleus connects to caudolateral areas of the rostral half of the medial accessory olive, whereas lateral areas project to more rostromedial parts. The most ventromedial part of the lateral cerebellar nucleus projects to the ventrolateral outgrowth. Adjacent medial, ventral, and caudal regions connect to the ventral leaf of the principal olive. The cerebellar origin of the projection to its dorsal leaf is located in lateral, dorsal, and rostral parts of the lateral cerebellar nucleus. The dorsolateral hump projects to the dorsomedial group of the rat inferior olive. Rostromedial projections to the dorsal accessory olive originate from the lateral part of the anterior interposed nucleus, whereas its medial parts project to more lateral and caudal regions of this olivary subnucleus. The dorsal fold of the dorsal accessory olive does not receive a projection from the cerebellar nuclei but from the lateral vestibular nucleus. No cerebellar projections were found to the dorsal cap. Relatively strong ipsilateral projections, which were the mirror images of the contralateral projections, were observed in the dorsomedial group, rostral medial accessory olive, and ventral leaf of the principal olive. When both the inferior olive and the cerebellar nuclei are considered as folded but continuous sheets of grey matter, the complete nucleo-olivary projection can be described as a simple transformation.

Distribution and brainstem origin of cholecystokinin‐like immunoreactivity in the opossum cerebellum

In order to determine the distribution of the peptide cholecystokinin (CCK) within the cerebellum and medullary precerebellar nuclei of the adult opossum, sections of these brain regions were processed for peroxidase-antiperoxidase immunohistochemistry. Within the inferior and superior cerebellar peduncles, fine-beaded fibers are evident and a beaded plexus of fibers is present in all the cerebellar nuclei. In the overlying cerebellar cortex, CCK-positive mossy fiber rosettes are present in all lobules, where their morphology varies from simple enlargements to more complex rosettes. However, their distribution varies particularly in vermal lobules II, III, VII, and IX where they are organized in parasagittal bands. Climbing fibers that are positive for CCK are present in very restricted areas of vermal lobules IV, VII, and VIII. After colchicine pretreatment, CCK-positive cell bodies are seen in restricted regions of the posterior interposed and fastigial nuclei as well as within several precerebellar nuclei known to give rise to mossy fibers. Such nuclei include the lateral cuneate nucleus, the nucleus prepositis hypoglossi, the nucleus reticularis lateralis, the nucleus raphe obscurus, the paramedian reticular nucleus, the nucleus reticularis gigantocellularis, and the medial vestibular nucleus. To localize the brainstem origin(s) of the CCK fibers in the cerebellum, a double-label paradigm employing a retrograde tracer and CCK immunohistochemistry was used. These experiments indicate that CCK mossy fibers originate primarily within the lateral cuneate nucleus, the perihypoglossal complex, and the lateral reticular nucleus. Some also originate within the medial vestibular nucleus and the nucleus reticularis gigantocellularis. In addition, double-labeled cell bodies are present within the caudal medial accessory inferior olive, the likely source of the CCK-positive climbing fibers. These data indicate that specific populations of climbing fibers and mossy fibers may utilize CCK to alter the firing rate of their target neurons.

Afferents of the caudal fastigial nucleus in a New World monkey (Cebus apella)

The afferents of the fastigial nucleus (FN) were studied in two capuchin monkeys (Cebus apella) one of which had received a unilateral injection of horseradish peroxidase in the caudal FN, and a second monkey which received a control injection that involved the lateral caudal FN but extended into the cerebellar white matter between the FN and posterior interposed nucleus (PIN). All of the sources of FN afferents were found to be labeled bilaterally. In addition to the restricted distribution of labeled Purkinje cells in lobules VI and VII of the posterior lobe vermis ("oculomotor vermis"), retrogradely labeled cells were present in the dorsolateral pontine nucleus (DLPN), dorsomedial pontine nucleus (DMPN), nucleus reticularis tegmenti pontis (NRTP), pontine raphe (PR), paramedian nucleus reticularis pontis caudalis (NRPC), nucleus prepositus hypoglossi (NPH), subnucleus b of the medial accessory olivary nucleus (sbMAO), and vestibular complex (VC). The second (control) injection appeared to confirm a proposed (Langer et al. 1985b) projection from the flocculus to the basal interstitial nucleus. The results are discussed in terms of the functional relationships of the FN to the frontal eye field and oculomotor-related brainstem structures involved in the production of saccadic and smooth pursuit eye movements.

The olivocerebellar projection to lobules I and II.

The olivocerebellar projection to lobules I and II was studied by means of retrograde transport from implants of the crystalline WGA-HRP complex. Retrogradely labelled neurons were found in the medial and dorsal accessory olives. Judged from the distribution of labelled cells, we conclude that parasagittally the olivocerebellar terminal zones A and B (i.e., the cerebellar cortical strips receiving axons from the olivary A and B regions) extend anteriorly into lobule Ia, whereas the fused olivocerebellar terminal C1/C3 zones reach lobule IIa. The olivocerebellar terminal C2 zone extends into lobule IIIa but not into lobule IIa. In lobule I the medial border of the B zone lies about 1 mm from the midline, in lobule II the B zone extends somewhat more medially. The lateral border of this zone is 1.9-2 mm from the midline. Compared to previous results, it appears that most of the Purkinje cells in lobule I projecting to the vestibular nuclei lie medially to the olivocerebellar terminal B zone.

Mesodiencephalic and cerebellar terminals terminate upon the same dendritic spines in the glomeruli of the cat and rat inferior olive: An ultrastructural study using a combination of [ 3H]-leucine and wheat germ agglutinin coupled horseradish peroxidase anterograde tracing

The mesodiencephalic and cerebellar afferents in the rostral medial accessory and principal olive of the cat and rat were studied following anterograde transport of tritiated leucine combined with anterograde transport of wheat germ agglutinin coupled horseradish peroxidase in the same animals. In all studied areas at least one-third of the labelled glomeruli appeared to contain both mesodiencephalic and cerebellar terminals. In many of these cases it was found that the terminals from both afferent systems contacted the same dendritic spines. Therefore, these olivary spines may be, as will be discussed, well suited for being involved in a timing process.

Coesxistence of corticotropin releasing factor and enkephalin in cerebellar afferent systems

In this study, we tested the hypothesis that corticotropin-releasing factor (CRF) and enkephalin (ENK) coexist within restricted brainstem-cerebellar circuits. Antisera for CRF and ENK were applied simultaneously or to serial sections of adult opossum brain and were processed for fluorescence or peroxidase, antiperoxidase immunohistochemistry, respectively. CRF and ENK were colocalized in 1) climbing fibers within the flocculus and in bilateral foci in the cerebellar vermis; 2) fibers with morphological characteristics of simple mossy fiber terminals or climbing fiber glomeruli that were located within the granule cell layer of the vermis underlying the foci of CRF/ENK-IR climbing fibers; 3) mossy fiber terminals within the flocculus; 4) neuronal perikarya in subnucleus A and C of the medial accessory olive and in the dorsal cap of Kooy, nuclei known to project as climbing fibers; and 5) cell bodies in nucleus prepositus, the subtrigeminal reticular nucleus, and the reticular formation, likely origins of CRF/ENK colocalized mossy fibers. The demonstration that single climbing and mossy fibers contain two peptides extends previous studies that have described chemical heterogeneity within cerebellar afferent pathways. Furthermore, these results support suggestions that this heterogeneity may provide a substrate for differential regulation of signal transduction by chemically coded cerebellar afferents.

Topographic and zonal pattern of olivocerebellar projection to the paramedian lobule in the rabbit: an experimental study with an HRP retrograde tracing method

Distribution of neurons in the inferior olive (IO) projecting to the paramedian lobule (PML) was studied in rabbits by means of retrograde transport of horseradish peroxidase (HRP). HRP was injected into various regions of different folia of the PML. Findings indicate zonal and to some extent topographic organization in olivocerebellar projections to PML folia. The neurons in corresponding areas of dorsal (dlPO) and ventral lamina (vlPO) of the principal olive (PO) project to composite zones D (D 1 + D 2) in sublobule f: medial, intermediate and lateral. In addition, the caudal part of the medial accessory olive (MAO) projects to the most laterally located zone (C 2-lateral) and the caudal part of the dorsal accessory olive (DAO) to the most lateral zone (C 3) in sublobule f. The middle part of the DAO sends projections to zone C 1 located medially in sublobules d-a. The rostral and caudal parts of the DAO send projections to zone C 3 in sublobules a and f, respectively. The rostral, middle and adjacent caudal parts of MAO, with no clear topographic organization, project to broad zone C 2 in sublobules e-b. The neurons in restricted areas of the caudomedial part of the dlPO and vlPO, probably intermingled with those supplying the composite medial zone D in sublobule f, project to sublobules e-b to terminate in zones D 1 and D 2, respectively.

Topographical organization of the olivocerebellar projection upon the posterior vermis in the rat

Projections from the medial accessory olive (MAO) to lobules VI, VII and VIII of the cerebellar vermis were investigated in the rat using the retrograde transport of wheatgerm agglutinin-conjugated horseradish peroxidase (WGA/HRP). When WGA/HRP was injected into the anterior half of lobule VI (VIa), retrogradely labeled neurons were found in a relatively narrow zone which traversed approximately the middle of the MAO and a portion of the nucleus β. In contrast, injection into the posterior half of lobule VI (VIb, c) resulted in labeling of neurons in the medial and lateral portions of the MAO and in the nucleus β. When HRP was confined to lobule VII, labeled neurons were found in the medial portion of the caudal MAO. No labeled neurons were found in the nucleus β. When the injection was restricted to lobule VIII, labeled neurons appeared along the caudal end of the MAO. There were many labeled neurons in the nucleus β. The central portion of the caudal MAO was free of labeled neurons unless the enzyme encroached upon a part of the anterior lobe. Thus, the projection areas to lobules VI–VIII formed a circle surrounding the free area and each of the lobules received fibers from a well-circumscribed portion: lobules VI–VIII received projections from the rostral, medial and caudal portion of the imaginary circle, respectively. The lateral portion was shared by the areas projecting to lobules VI and VIII. In addition, these lobules received projections also from an overlapping area in the nucleus β.

Subcortical connections of frontal ‘oculomotor’ areas in the cat

The subcortical connections of the frontal ‘oculomotor’ areas in the medial wall of the hemisphere under the cruciate sulcus (CRUo), the medial (PREo-m) and the lateral banks (PREo-l) of the presylvian sulcus of cats were investigated using WGA-HRP tracing combined with electrophysiologic techniques. Two main modes were identified; one was the common connections to the same targets in the cortico-thalamo-cortical, cerebro-cerebellar and cortico-tectal pathways, and the other was the individual connections to unique portions of the saccade-generating centers in the brainstem. The common reciprocal connections were found in the ventral anterior-ventral lateral complex, principal ventromedial nucleus, rostral intralaminar nuclei, centromedian- parafascicular complex, lateral posterior nucleus, and suprageniculate nucleus. The common efferent projections were in the subthalamic nucleus, lateral habenular nucleus, pretectal nucleus, posterior commisure nucleus, nucleus of Darkschewitsch, pontine nucleus, nucleus reticularis tegmenti pontis, medial accessory inferior olive, and the superior colliculus. The CRUo projected into the ipsilateral field of Forel and paramedial pontine reticular formation (PPRF), the PREo-1 projected into the contralateral dorsomedial medullary reticular formation, and the PRE0-m projected into the ipsilateral medullary reticular formation and there was only a small degree of projection into the central portion between the abducens nerve rootlets.

Spino‐olivary termination on spines in cat medial accessory olive

Three functional regions of the inferior olive, the caudal medial accessory olive (cMAO) and the caudal and rostral dorsal accessory olive (DAO) receive input from the spinal cord. The present study determined how spinal inputs to cMAO interact with olivary neurons. These inputs were labeled by injections in cat lumbosacral of wheat germ agglutinin conjugated to horseradish peroxidase. The tracer was visualized with tetramethylbenzidine. The morphology of the labeled spino-olivary terminals and the relationship between these terminals and postsynaptic elements were determined. Spino-olivary terminals in cMAO displayed the morphological characteristics classically associated with excitatory synapses. Almost three quarters synapsed on spines, most of which contacted other spines, forming spine clusters. The majority of postsynaptic spines also received convergent input from apparently excitatory, nonlumbosacral afferents. This postsynaptic organization provides several possible benefits for the putative role of cMAO in the control of posture. An earlier study demonstrated that in DAO, almost three quarters of lumbosacral, spino-olivary terminals synapse on dendrites (Molinari: Neuroscience 27:425-435, 1988). Thus, lumbosacral afferents appear to differ fundamentally in the way in which they interact with neurons in cMAO and DAO. These results suggest that the way information is processed may be as important in determining the functional differences between olivary regions as what information is received.

Anterograde tracing of the rat olivocerebellar system with Phaseolus vulgaris leucoagglutinin (PHA-L). Demonstration of climbing fiber collateral innervation of the cerebellar nuclei.

The olivocerebellar climbing fiber system was investigated in the rat with anterograde Phaseolus vulgaris leucoagglutinin (PHA-L) tracing. The specific objective of the study was to find morphological evidence of climbing fiber collaterals innervating the cerebellar nuclei. Small iontophoretic injections of PHA-L were placed in different parts of the inferior olivary complex, and labelled olivocerebellar fibers could be traced to their termination as climbing fibers in sagittal zones of the contralateral cerebellar cortex. Reaching the cerebellum via the restiform body, the labelled olivocerebellar axons entered the deep cerebellar white matter anterior to the cerebellar nuclei. Most of these thicker, nonterminal axons continued dorsally around the nuclei, but some ran through them. Bundles of fibers could be followed into the folial white matter toward their cortical zones of termination. Depending on which part of the olivary complex that was injected with PHA-L, labelled axons were seen to converge on different regions of the cerebellar nuclei, where dense plexuses of thin varicose terminal fibers appeared. Quantitative estimates of the innervation ranged from 1.7 to 4.3 million boutons per mm3 in the fastigial (FN), interposed, and main parts of the lateral cerebellar (LCN) nuclei, whereas the parvicellular portion of LCN demonstrated 15-20 million varicosities per mm3. Frequently, thicker olivocerebellar axons, which seemed directed toward the cerebellar cortex, were seen to send a fine collateral branch toward these areas of nuclear innervation. As controls, PHA-L was injected into the degenerated olivary complex of 3-acetylpyridine-treated rats. Neither cortical climbing fiber terminals nor nerve terminal plexuses in the nuclei appeared in these experiments. In cases with injection sites extending into the reticular formation, substantial mossy fiber labelling was present bilaterally in the cortex, but the cerebellar nuclei were devoid of labelled innervation or demonstrated only a few larger diameter fibers. The projection of the inferior olivary complex to the cerebellar nuclei was strictly topographically organized and agreed in principle with the organization described in the cat by Groenewegen et al. ('79). The caudal medial accessory olive (MAO) projected to FN, the rostral MAO to the posterior interposed nucleus (NIP), the rostral part of the dorsal accessory olive (DAO) to the anterior interposed nucleus (NIA), and the principal olive (PO) to LCN.(ABSTRACT TRUNCATED AT 400 WORDS)

Organization of cerebral cortico-olivary projections in the rat

Injection of wheatgerm agglutinin-conjugated horseradish peroxidase into electrophysiologically identified portions of the rodent sensorimotor and medial frontal cortex revealed anterograde-labeled projections to the inferior olivary complex. Forelimb motor cortical and supplementary motor cortical regions were found to project predominantly to the principal olive while forelimb sensory cortex fibers distributed mainly within the dorsal accessory olive. Forelimb cortical fibers (both sensory and motor) terminated more rostrally and medially in the medial and dorsal accessory olives than did hindlimb projections. Medial frontal cortex projected to the beta subnucleus, the rostral medial accessory olive (especially the ventral aspect), and to the ventrolateral outgrowth of the dorsal cap. These findings indicate a more extensive origin for cortico-olivary projections than previously described for rats or cats and they demonstrate a rather precise topography for olivary projections from various cortical regions.

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.

Axonal branching of the olivocerebellar projection in the rat: A double‐labeling study

Collateralization of olivocerebellar (climbing) fibers was studied in the rat by means of the fluorochrome double-labeling technique. Most of the olivocerebellar projection is crossed except for a minimal ipsilateral component which arises from the most rostal part of the inferior olivary nucleus (ION). ION neurons in the caudolateral part of the medial accessory olive (MAO) and the dorsal accessory olive (DAO) give off axons that branch to supply both hindlimb areas of the contralateral cerebellar cortex, i.e., the rostral anterior lobe and the caudal paramedian lobule. In addition, neurons in the middle one-third of the contralateral MAO and DAO send axons that divide to terminate in both the caudal part of the anterior lobe and the rostral part of the paramedian lobule (forelimb receiving areas). Neurons within the caudal part of the MAO, the lateral part of the DAO, the ventral lamella of the principal olive (PO), and the dorsomedial cell column (DMCC) send axonal branches that terminate within at least two different areas of the same sagittal zones throughout the contralateral cerebellar cortex. Thus, the ION contains specialized cells that provide a divergence of integrated information from the ION to at least two cerebellar regions.

Ultrastructural study of the GABAergic, cerebellar, and mesodiencephalic innervation of the cat medial accessory olive: Anterograde tracing combined with immunocytochemistry

The rostral medial accessory olive (MAO) of the cat was studied by using an ultrastructural technique combining wheat germ agglutinin-coupled horseradish peroxidase (WGA-HRP) anterograde tracing and postembedding GABA immunocytochemistry. One group of cats received a WGA-HRP injection in the posterior interposed nucleus of the cerebellum and another group received an injection in the nucleus of Darkschewitsch. Based on differences in their morphology three types of GABAergic and three types of nonGABAergic terminals were observed. One type of GABAergic terminal was often GABA/WGA-HRP double-labeled in the cerebellar experiments, and one type of nonGABAergic terminal was often WGA-HRP-labeled in the mesodiencephalic experiments. Following injections of WGA-HRP in the cerebellar nuclei virtually all WGA-HRP-labeled terminals were GABA positive. Quantification of these GABA/WGA-HRP-double-labeled terminals showed that 1) 30% of the GABAergic terminals randomly selected from the entire neuropil were double-labeled, 2) 13% of the GABAergic terminals adjacent to perikarya were double-labeled, and 3) 34% of the GABAergic terminals strategically located next to both of the dendritic elements linked by a gap junction were double-labeled. Statistical analysis of the above data showed that significantly fewer GABAergic terminals adjacent to perikarya were double-labeled (P less than .001) than would be expected from the double-labeled proportion of the randomly selected GABAergic terminals. Following injection of WGA-HRP in the nucleus of Darkschewitsch, all WGA-HRP-labeled terminals were GABA-negative. Quantification of these terminals showed that 1) 26% of the randomly selected nonGABAergic terminals were WGA-HRP labeled, 2) 20% of the nonGABAergic terminals adjacent to perikarya were WGA-HRP labeled, and 3) 23% of the nonGABAergic terminals strategically located next to a gap junction were WGA-HRP labeled. No significant differences were found among these populations. Quantification of terminals of both groups of experiments mentioned above showed that GABAergic terminals composed 1) 38% of the randomly selected terminals, 2) 64% of the terminals apposed to perikarya, and 3) 53% of the terminals strategically located next to gap junctions. Statistical analysis of the above data showed that significantly more GABAergic terminals were located adjacent to perikarya (P less than .001) and strategically next to a gap junction (P less than .05) than would be expected from the random GABAergic innervation. The above findings of the GABAergic, cerebellar, and mesodiencephalic input are discussed with regard to their functional role in the neuronal circuitry of the ros