Afferent projection from reticular nuclei, inferior olive and cerebellum to lateral vestibular nucleus of the cat as demonstrated by horseradish peroxidase

Afferent connection to lateral vestibular nucleus (LVN) was examined using retrograde transport of horseradish peroxidase (HRP). When HRP was microiontophoretically applied to the immediate vicinity of the LVN neuron, which monosynaptically fired spike upon VIIIth cranial nerve stimulation, HRP-labelled cells were observed in the ipsilateral lateral reticular nucleus, bilateral gigantocellular nucleus, and contralateral dorsal cap and beta-nucleus of inferior olive in addition to various parts of cerebellum.     

Cortical and brain stem afferents to the ventral thalamic nuclei of the cat demonstrated by retrograde axonal transport of horseradish peroxidase

After horseradish peroxidase (HRP) injections into various parts of the ventral thalamic nuclear group and its adjacent areas, the distribution of labeled neurons was compared in the cerebral cortex, basal ganglia, and the brain stem. The major differences in distribution patterns were as follows: Injections of HRP into the lateral or ventrolateral portions of the ventroanterior and ventrolateral nuclear complex of the thalamus (VA-VL) produced retrogradely labeled neurons consistently in area 4 gamma (lateral part of the anterior and posterior sigmoid gyri, lateral sigmoid gyrus and the lateral fundus of the cruciate sulcus), the medial division of posterior thalamic group (POm), suprageniculate nucleus (SG) and anterior pretectal nucleus ipsilaterally, and in the nucleus Z of the vestibular nuclear complex bilaterally. Injections into the medial or dorsomedial portion of the VA-VL resulted in labeled neurons within the areas 6a beta (medial part of the anterior sigmoid gyrus), 6a delta (anterior part of ventral bank of buried cruciate sulcus), 6 if. fu (posterior part of the bank), fundus of the presylvian sulcus (area 6a beta), medial part of the nucleus lateralis posterior of thalamus and nucleus centralis dorsalis ipsilaterally, and in the entopeduncular nucleus (EPN) and medial pretectal nucleus bilaterally. Only a few neurons were present in the contralateral area 6a delta. After HRP injections into the ventral medial nucleus (VM), major labeled neurons were observed in the gyrus proreus, area 6a beta (mainly in the medial bank of the presylvian sulcus), and EPN ipsilaterally, and in the medial pretectal nucleus and substantia nigra bilaterally. Following HRP injections into the centre médian nucleus (CM), major labeled neurons were found in the areas 4 gamma, 6a beta, and the orbital gyrus ipsilaterally, and in the EPN, rostral and rostrolateral parts of the thalamic reticular nucleus, locus ceruleus, nucleus reticularis pontis oralis et caudalis and nucleus prepositus hypoglossi bilaterally. The contralateral intercalatus nucleus also possessed labeled neurons. With HRP injections into the paracentral and centrolateral nuclei, labeled neurons were observed in the gyrus proreus and the cortical areas between the caudal presylvian sulcus and anterior rhinal sulcus ipsilaterally, and in the nuclei interstitialis and Darkschewitsch bilaterally. Minor differences in the distribution pattern were observed in the superior colliculus, periaqueductal gray, mesencephalic and medullary reticular formations, and vestibular nuclei in all cases of injections.     

Projections from the spinal and the principal sensory nuclei of the trigeminal nerve to the cerebellar cortex in the cat, as studied by retrograde transport of horseradish peroxidase.

Projections from the spinal (Vsp) and the principle sensory (Vp) nuclei of the trigeminal nerve to the cerebellar cortex were studied by means of retrograde transport of horseradish peroxidase in the cat. Neurons projecting to the simple lobule and the dorsal part of the paramedian lobule (PMD) were located mainly in the dorsal part of the nucleus interpolaris (Vi) and of the caudal one third of the nucleus oralis (Vo) and in the rostralmost part of the Vo. Neurons projecting to the medial part of the posterior folia of crus II (crus IIp) were located in the dorsal to ventral parts of the Vi and of the caudal one third of the Vo and in the rostralmost part of the Vo, while those projecting to the lateral part of crus IIp were confined to the ventral part of the Vi and of the caudal one third of the Vo. Neurons of the Vp also projected to all of these cortical areas. They were relatively confined to the ventral part of this nucleus. Thest trigeminocerebellar projections were exclusively ipsilateral to the cell origin. There were sparse projections from the Vi and Vo to lobules V to VIIIa. In addition, a small group of neurons in the subnucleus magnocellularis of the nucleus caudalis of the Vsp also projected to the above cortical areas. No projections were, however, observed to the anterior portion of the anterior lobe, crus I, the anterior folia of crus II, paraflocculus, flocculus and the ventral part of the PMD. The majority of these cerebellar projection neurons were medium-sized and triangular, fusiform or ovoid in shape. There were small neurons of similar types and large multipolar neurons as well.     

Projection from the vestibular nuclei to the inferior olive in the cat: An autoradiographic and horseradish peroxidase study

Injections of horseradish peroxidase (HRP) in the inferior olive of 13 cats resulted in the labeling of neurons in the medial (MVN) and descending vestibular nuclei (DVN) as well as in subgroups g, p.h., s.v., x and z. The contribution of groups p.h. and z. could not be definitively established. Injections of triated leucine in the vestibular nuclei and adjacent reticular formation in 4 cats indicated that the previously identified vestibular nuclei and subgroups project bilaterally to the dorsomedial cell column and ipsilaterally to subnucleus beta of the inferior olive. In view of the known topography of olivocerebellar projections, the present results suggest that climbing fiber-mediated vestibular information may influence the uvula and the fastigial nucleus.     

The afferent projections to the centrum medianum of the cat as demonstrated by retrograde transport of horseradish peroxidase

The afferent projections of nucleus centrum medianum (CM) of the thalamus were studied, in the cat, by means of retrograde transport of electrophoretically ejected horseradish peroxidase. Several variations of method — survival time, fixatives, substrates, etc. — were tried to improve the amount of visible reaction product.Labeled neurons were localized primarily in two categories of nuclei in the brain. The first consisted of structures making up or closely related to the basal ganglia: the entopeduncular nucleus, the pars reticulata of the substantia nigra, and motor cortex. The second category was made up of nuclei closely related to postural and orienting functions: the deep layers of the superior colliculus ipsilaterally, and the medial and lateral vestibular nuclei bilaterally. Other nuclei containing retrogradely labeled neurons were the periaqueductal gray and locus coeruleus. Brain stem reticular projections were sparse and widely scattered. These results identify CM as an important element in the loop system linking medial thalamus and neostriatum; the probable attention and orientation related functions of this system are discussed.     

Nigrostriatal projections in the rat as demonstrated by retrograde transport of horseradish peroxidase. I. Projections to the rostral striatum

The distribution of nigral neurons projecting to the rostral part of the striatum was studied in 12 rats using the horseradish peroxidase or the wheatgerm lectin bound horseradish peroxidase labelling techniques. Labelled neurons localized in the substantia nigra pars compacta (SNc) were demonstrated throughout the anteroposterior extent of the nucleus. Most of the labelled neurons were localized in the medial half of the SNc, predominantly in its basal part. Labelled neurons localized in the substantia nigra pars reticulata (SNr) predominated in the caudal half of the nucleus where they were found in its medial, central and lateral parts. A quantitative evaluation of the shape and size of the labelled neurons showed no statistically significant differences between the SNc and SNr as to the shape of the labelled perikarya. In contrast, nigrostriatal neurons in the SNc were found to have larger perikarya than their counterparts in the SNr.     

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.     

Projections of lamprey spinal neurons determined by the retrograde axonal transport of horseradish peroxidase.

The spinal cords of larval sea lampreys (Petromyzon marinus) and adult river lampreys (Ichthyomyzon unicuspis) were injected with horseradish peroxidase through a transection 1 cm caudal to the last gill. Some animals also had a spinal hemisection 1 cm caudal to the injection. After recovery periods of 1 to 52 days, the spinal cords were treated with diaminobenzidene and hydrogen peroxide, and the projections of various cell types determined in wholemount slides. From these observations the following conclusions were drawn. Most dorsal cells (primary sensory cells) are bipolar with a long rostral projection and a short caudal projection of no more than 5-10 mm. Both processes travel in the ipsilateral dorsal column. Their peripheral processes enter the dorsal roots as branches of their central axons. Some dorsal cells send processes out three or more dorsal roots both rostral and caudal to the cell body. Myotomal motoneurons have characteristic locations in the medial gray column and send prominent transversely oriented dendrites into the lateral columns. A few motoneurons are unusually large. In addition to giant interneurons the majority of smaller rostrally projecting interneurons also have decussating axons. A recently described cell type, the oblique bipolar cell, appears to have an exclusively crossed rostral projection. Although most edge cells project rostrally, as many as 20% may have a caudal projection or both rostral and caudal projections. Edge cells project equally to the ipsilateral and contralateral spinal hemicord, but their processes do not extend more than about 18 mm in sea lamprey larvae and 37 mm in adult river lampreys. Lateral cells project exclusively to the ipsilateral caudal hemicord. A few cells which resemble lateral cells in location and in possessing large lateral dendrites, project rostrally. However, these have atypical morphologic features which probably distinguish them from true lateral cells. Thus far, regardless of cell type, all decussating axons seem to pass ventral to the central canal, while decussating medial dendrites pass dorsally.     

Dorsal column nuclei projections to the cerebellar cortex in cats as revealed by the use of the retrograde transport of horseradish peroxidase.

The existence of a cerebellar projection from the dorsal column nuclei (gracile and cuneate nuclei, DCN) has been proposed on electrophysiological grounds but questioned when studied with neuroanatomical techniques. The retrograde transport of horseradish peroxidase (HRP) has been used for the present study and provides anatomical evidence of a DCN-cerebellar pathway. In adult cats, 1 to 6 mul of 30% HRP were injected in pars intermedia of the anterior lobe (lobules IV-V), in paramedial lobule and in vermis of the anterior (lobules IV-V) and of the posterior lobe (lobule VII). After survival of 24 to 48 hours, all animals were perfused with a double aldehyde mixture and serial 40 mu sections through the medulla oblongata were incubated for visualization of HRP. In all cases, medullary nuclei known to project to the injected cortical regions of the cerebellum contained HRP-positive neurons mainly ipsilateral to the injection (e.g., external cuneate nucleus) or mainly contralateral to it (e.g., inferior olivary complex). Following ipsilateral injections in either the paramedian lobule or the pars intermedia, HRP-positive neurons in the cuneate nucleus were concentrated in its rostral portion where multipolar cells with radiating dendrites predominate. In contrast, none of the clusters region, in the caudal part of the cuneate nucleus, displayed HRP-positive granules. In cases in which the anterior vermis was injected a few labelled cells were present in the rostral part of the gracile nucleus but not in the clusters region of this nucleus. No labelling of DCN neurons was evident after posterior vermis injection. To compare the distribution of cells contributing to the DCN-cerebellar pathway with that of thalamic relay cells in the DCN, 0.5 to 3 mul of 30% HRP were injected in the nucleus ventralis posterolateralis of the thalamus in another series of cats. Contralateral to the thalamic injection, labelled cells were concentrated in the clusters region of the gracile and cuneate but rostrally in these nuclei they were scattered among unlabelled neurons. The preferential location in the DCN of cells which project to the cerebellum and of cells which project to the thalamus stresses the heterogeneous organization of these nuclei along the rostrocaudal axis. Further, the results indicate that regions of the DCN which have been distinguished on the basis of cytoarchitectonics (Kuypers and Tuerk, '64) and of afferents (Rustioni, '73, '74) differ also in their efferent projections.     

Mesencephalic and diencephalic afferents to the superior colliculus and periaqueductal gray substance demonstrated by retrograde axonal transport of horseradish peroxidase in the cat

The mesencephalic and diencephalic afferent connections to the superior colliculus and the central gray substance in the cat were examined by means of the retrograde transport of horseradish peroxidase (HRP). After deep collicular injections numerous labeled cells were consistently found in the parabigeminal nucleus, the mesencephalic reticular formation, substantia nigra pars reticulata, the nucleus of posterior commissure, the pretectal area, zona incerta, and the ventral nucleus of the lateral geniculate body. A smaller number of cells was found in the inferior colluculus, the nucleus of the lateral lemniscus, the central gray substance, nucleus reticularis thalami, the anterior hypothalamic area, and, in some cases, in the contralateral superior colliculus, Forel's field, and the ventromedial hypothalamic nucleus. Only the parabigeminal nucleus and the pretectal area showed labeled cells following injections in the superficial layers of the superior colliculus. In the cats submitted to injections in the central gray substance, labeled cells were consistently found in the contralateral superior colliculus, the mesencephalic reticular formation, substantia nigra parts reticulata, zona incerta and various hypothalamic areas, especially the ventromedial nucleus. In some cases, HRP-positive cells were seen in the nucleus of posterior commissure, the pretectal area, Forel's field, and nucleus reticularis thalami. A large injection in the mediodorsal part of the caudal mesencephalic reticular formation, which included the superior colliculus and the central gray substance, resulted in numerous labeled cells in nucleus reticularis thalami. The findings are discussed with respect to the suggested functional division of the superior colliculus into deep and superficial layers. Furthermore, the possible implications of labeled cells in zona incerta and the reticular thalamic nucleus are briefly discussed.     

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.     

Spinal projections from the nucleus locus coeruleus and nucleus subcoeruleus in the cat and monkey as demonstrated by the retrograde transport of horseradish peroxidase

The rostral pons of the cat and rhesus monkey were examined for the presence of labeled cells following injections of horseradish peroxidase (HRP) into the lumbar spinal cord. Labeled cells were found in the ipsilateral dorsolateral pontine tegmentum and in the contralateral ventrolateral pontine reticular formation. In both the cat and monkey, labeled cells were located in the nucleus locus coeruleus, nucleus subcoeruleus, in or near the Kölliker-Fuse nucleus, and in the ventral part of the lateral parabrachial nucleus. There is a striking similarity between the distribution of HRP-labeled cells in the dorsolateral pontine tegmentum of the cat and monkey and that of catecholamine-containing cells observed in this area in previous studies.     

The inferior olivary connections to the cerebellum in the rat studied by retrograde axonal transport of horseradish peroxidase

Olivocerebellar projections were investigated in the rat using retrograde axonal transport of horseradish peroxidase. Discrete cell groups of the inferior olive were labelled, subsequent to injections in the paravermal region, the vermis, or the caudolateral hemisphere. Injections in the midrostrocaudal third of the paravermal area resulted in labelling of cells in the medial accessory olive (MAO), in cell group “b” at caudal levels, and in its lateral portion at mid-rostrocaudal levels. The rostral pole of the principal olive (PO), the dorsal accessory olive (DAO), and the dorsomedial cell column, were heavily labelled. By comparison, caudal paravermal injections resulted in labelling in the medial part of the mid-rostrocaudal levels of the MAO, but not in its caudal portion. The PO lamellae were labelled in their lateral half, excluding the lateral bend connecting them. Injections slightly lateral within this paravermal area gave no caudal MAO labelling, but did label cells in segments of both PO lamellae, medial to those in the previous case. From vermal injections, cell groups “b” and “c” of the caudal MAO were labelled, but no labelled cells were present in the PO. Subsequent to injections in the paramedian lobule, cells in the dorsal lamella of the PO were labelled. No cells of the MAO were labelled. These results are discussed in terms of specific labelling patterns and the general concepts of organization presently held for the Olivocerebellar system.