Neuroactive amino acids in the area postrema. An immunocytochemical investigation in rat with some observations in cat and monkey (Macaca fascicularis).

The localization of five neuroactive amino acids in the rat area postrema was studied by postembedding immunocytochemistry in semithin and ultrathin sections. Antisera to GABA, glycine, glutamate and aspartate produced labelling of cells that were identified as neurons in the electron microscope. GABA-like and glycine-like immunoreactivities occurred in about 20% and 60% of the neurons, respectively, and a minor proportion of the cells displayed both immunoreactivities, suggesting a cellular colocalization of GABA and glycine.

Metabolic compartmentation of glutamate and glutamine: morphological evidence obtained by quantitative immunocytochemistry in rat cerebellum.

An electron microscopic, double-labelling immunocytochemical procedure was used to assess the level of fixed glutamate and glutamine in different cell profiles in ultrathin sections of rat cerebellar cortex. The procedure was based on sequential immunolabelling with two rabbit antisera, using gold particles of different sizes as markers and formaldehyde vapour as a means to avoid interference between the two incubations. Model sections containing a series of known concentrations of the respective amino acids (aldehyde--fixed to rat brain protein) were incubated together with the tissue material.

An electron microscopic, immunogold analysis of glutamate and glutamine in terminals of rat spinocerebellar fibers.

A semiquantitative, electron microscopic immunocytochemical procedure based on the use of colloidal gold particles as markers was employed to analyze the subcellular distribution of glutamate and glutamine, a major glutamate precursor, in a subpopulation of spinocerebellar mossy fiber terminals. These terminals were identified by anterograde transport of a horseradish peroxidase-wheat germ agglutinin conjugate, injected in the thoracic spinal cord. Gold particles signalling glutamate-like immunoreactivity were enriched over clusters of synaptic vesicles relative to organelle-free cytoplasmic matrix, and there was a strong positive correlation between gold particle and synaptic vesicle densities (correlation coefficient 0.

Afferent connections to the abducent nucleus in the cat.

The afferent connections to the abducent nucleus in the cat were studied by means of retrograde transport of WGA-HRP after implantations of the tracer in crystalline form. Retrogradely labelled cells were found bilaterally in the medial and descending vestibular nuclei, mainly in their ventral and medial portions, in the rostral part of the ipsilateral gigantocellular reticular nucleus, in the medial part of the contralateral caudal pontine reticular nucleus and bilaterally in the oculomotor nucleus, mainly in its dorsolateral division. Some labelled cells were also found bilaterally in the mesencephalic reticular formation, the periaqueductal grey and the nucleus of the trapezoid body.

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.

Aspartate-like and glutamate-like immunoreactivities in the inferior olive and climbing fibre system: a light microscopic and semiquantitative electron microscopic study in rat and baboon (Papio anubis).

A post-embedding immunogold procedure was used to analyse, in a semiquantitative manner, the distributions of aspartate-like and glutamate-like immunoreactivities in the inferior olive and climbing fibre system in rats and baboons. The neurons in the inferior olive were uniformly labelled for aspartate as well as glutamate, indicating a 100% co-localization of these two amino acids in the cell bodies. The level of glutamate-like immunoreactivity in the climbing fibre terminals was similar to that in the parent cell bodies, as judged by a computer-assisted calculation of gold particle densities.

GABA, glycine, aspartate, glutamate and taurine in the vestibular nuclei: an immunocytochemical investigation in the cat.

The distributions of five amino acids with well-established neuroexcitatory or neuroinhibitory properties were investigated in the feline vestibular complex. Consecutive semithin sections of plastic-embedded tissue were incubated with antisera raised against protein-glutaraldehyde conjugates of GABA, glycine, aspartate, glutamate and taurine. This approach allowed us to study the relative densities of the different immunoreactivities at the level of individual cell profiles.

Demonstration of GABA immunoreactive cells in the inferior olive of baboons (Papio papio and Papio anubis).

The distribution of gamma-aminobutyric acid (GABA)-like immunoreactivity was studied in semithin sections through the inferior olivary complex in two baboon species. About 5% of the olivary neurons were GABA-immunoreactive. The GABA-immunoreactive neurons differed from the large majority of olivary neurons by their smaller size and their lower contents of aspartate, as judged by analysis of alternate sections labelled with an aspartate antiserum.

The interconnection between the vestibular nuclei and the nodulus: a study of reciprocity.

The reciprocal connections between the nodulus and the vestibular and perihypoglossal nuclei in the cat have been studied by anterograde and retrograde transport after implants of crystalline wheatgerm agglutinin-horseradish peroxidase complex (WGA-HRP) restricted to one or two nodular folia. The findings supplement the previous study by Epema et al. (Neurosci.

The olivonodular projection: a re-examination based on folial cerebellar implants.

In 5 cats implants of crystalline wheat germ agglutinin-horseradish peroxidase complex were placed in one or two folia of the nodulus with no contamination of adjacent cerebellar lobules. In the inferior olive only the dorsal cap with the adjacent ventrolateral outgrowth and the caudalmost part of nucleus beta were found to project to the nodulus. Negative findings were consistently made in all other parts of the inferior olive, some of which have previously been described to contribute to the olivonodular projection.

Cerebellar nuclear afferents--where do they originate? A re-evaluation of the projections from some lower brain stem nuclei.

Cerebellar nuclear afferents from some caudal brain stem nuclei in the cat were studied by means of retrograde transport after implantation of the wheat germ agglutinin-horseradish peroxidase complex in crystalline form in the cerebellar nuclei. The findings give evidence that projections to the cerebellar nuclei from certain nuclei of the reticular formation proper (e.g.

Is there a reciprocal connection between the red nucleus and the interposed cerebellar nuclei? Conclusions based on observations of anterograde and retrograde transport of peroxidase-labelled lectin in the same animal.

The rubrointerposital projection was studied in cats where wheat germ agglutinin-horseradish peroxidase (WGA-HRP) was implanted in various parts of the interposed nuclei. No retrogradely labelled rubral cells were observed following implantations in the posterior interposed nucleus, and only very few such cells were identified in the contralateral red nucleus after implantations restricted to the anterior interposed nucleus with no contamination of cerebellar white matter or cortex. However, when WGA-HRP was delivered by a pressure injection, which in addition to the anterior interposed nucleus included the adjacent white cerebellar matter along the needle track and the overlying cortex, many retrogradely labelled cells were present contralaterally in the magnocellular red nucleus, with some also found in its rostral parvicellular part.

Do pontocerebellar mossy fibres give off collaterals to the cerebellar nuclei? An experimental study in the cat with implantation of crystalline HRP-WGA.

In 15 cats with implantations of crystalline HRP-WGA in the cerebellar nuclei and tetramethylbenzidine histochemistry, the pontine nuclei were carefully examined for presence of retrogradely labelled cells. Findings in the nucleus reticularis tegmenti pontis and the inferior olive, both known to project to the cerebellar nuclei, served as controls for effectiveness of uptake and transport. After implantations restricted to the lateral cerebellar nucleus in 5 cats altogether two labelled cells were found in the contralateral pontine nuclei in regions receiving afferents from the lateral nucleus.

The cerebellar nucleo-olivary and olivocerebellar nuclear projections in the cat as studied with anterograde and retrograde transport in the same animal after implantation of crystalline WGA-HRP. III. The interposed nuclei.

The bidirectional connections between the inferior olive and the cerebellar nuclei were investigated by means of anterograde and retrograde transport after implantation of crystalline wheat germ agglutinin-horseradish peroxidase complex in the interposed nuclei. The projections from the interposed nuclei to the inferior olive show a detailed topical arrangement. The main projection from the anterior interposed nucleus reaches the rostral two thirds of the dorsal accessory olive, while the main projection from the posterior interposed nucleus reaches the rostral half of the medial accessory olive.

Ultrastructure of spinal efferents to the lateral reticular nucleus: an EM study using anterograde transport of WGA-HRP complex.

Anterograde transport of lectin-conjugated horseradish peroxidase and subsequent incubation with tetramethylbenzidine were employed to label the spinal terminals within the feline lateral reticular nucleus (NRL) for ultrastructural identification. Quantitative studies demonstrated that compared to the unlabelled terminals the spinal boutons were more than twice as large and contained fewer synaptic vesicles. Most of the synaptic contacts of the labelled terminals were located on dendritic shafts but contacts on dendritic spines as well as perikarya were also present.

The origin and termination of the dentatorubral fibres in the cat as studied with retrograde and anterograde transport of peroxidase labelled lectin.

The dentatorubral projection in the cat was studied by means of retrograde and anterograde transport after implantations of crystalline wheat germ agglutinin-horseradish peroxidase (WGA-HRP) in the red and dentate nuclei, respectively. Our retrograde transport findings give evidence that rubral afferents originate in the contralateral dentate nucleus, mainly dorsally in its central part. The dentate implantations (with no contamination of the adjacent interposed nuclei) show that in the cat the great majority of the dentatofugal fibres leave the ipsilateral brachium conjunctivum to decussate below the red nucleus.

The cerebellar nucleo-olivary and olivo-cerebellar nuclear projections in the cat as studied with anterograde and retrograde transport in the same animal after implantation of crystalline WGA-HRP. I. The dentate nucleus.

The implantation technique described by Mori et al. has been modified for the implantation of crystalline wheat germ agglutinin-horseradish peroxidase (WGA-HRP) complex. This method permits a detailed analysis of the afferent and efferent connections of the cerebellar nuclei without the complication of uptake and transport of the tracer into passing fibres.

On the projections from the vestibular and perihypoglossal nuclei to the spinal trigeminal and lateral reticular nuclei in the cat.

The projection from the vestibular and perihypoglossal nuclei to the spinal trigeminal and lateral reticular nuclei has been studied in cats where the wheat germ agglutinin-horseradish peroxidase complex has been used as a retrograde tracer. All injections were made at the level of the caudal pole of the inferior olive. The medial and descending vestibular, and the perihypoglossal nuclei were found to project to the spinal trigeminal nucleus.

Cerebellar nuclear afferents from feline hypothalamus demonstrated by retrograde transport after implantation of crystalline wheat germ agglutinin-horseradish peroxidase complex.

A hypothalamo-cerebellar projection has recently been discovered in the cat, and its presence has been verified in other mammalian species. Due to methodological limitations only hypothalamic projections to cerebellar cortex had been demonstrated. By means of modification of the new implantation technique described by Mori et al.

A projection from the periaqueductal grey to the lateral reticular nucleus in the cat.

A projection from the periaqueductal grey (PAG) to the lateral reticular nucleus (NRL) in the cat was demonstrated by means of retrograde transport of the wheat germ agglutinin-horseradish peroxidase complex. The connection has its main origin ipsilaterally in the ventral part of the caudal PAG, but scanty projections from other parts of the PAG were also found. The neurons projecting to the NRL are of varying shapes and sizes, but most cells have a maximum diameter of less than 20 micron.

The cerebellar nucleo-olivary and olivo-cerebellar nuclear projections in the cat as studied with anterograde and retrograde transport in the same animal after implantation of crystalline WGA-HRP. II. The fastigial nucleus.

The bidirectional connections between the inferior olive and the fastigial nucleus were studied by means of anterograde and retrograde transport after implantation of crystalline wheat germ agglutinin-horseradish peroxidase (WGA-HRP) complex into the fastigial nucleus. The fastigio-olivary fibres terminate in the caudal half of the medial accessory olive, nucleus beta and the dorsal cap, and the olivo-fastigial projection has its origin within the same olivary regions. A topical arrangement is indicated for both pathways.

On the morphology of the mesencephalic trigeminal cells. New data based on tracer studies.

Injections of free horseradish peroxidase in the masticatory muscles of the cat resulted in retrograde labeling of not only large and small so-called pseudounipolar cells but also of multipolar neurons within the ipsilateral mesencephalic trigeminal nucleus. The latter cell type was present only in the pontine part of the nucleus, and usually more faintly labeled than the other cells. Several of the so-called pseudounipolar cells showed cell processes similar to dendrites of other cells, a finding indicating that the afferent connections of these cells are much more complex than hitherto assumed.

An ipsilateral projection from the red nucleus to the lateral reticular nucleus in the cat.

Injections of the wheat germ agglutinin--horseradish peroxidase complex into the lateral reticular nucleus reveal that in addition to the well known contralateral rubroreticular connection, there is also a small but clear cut ipsilateral projection. Cells of various sizes participate in this ipsilateral pathway, and the retrogradely labelled neurons lie dispersed throughout the entire red nucleus.

The medullary projection from the mesencephalic trigeminal nucleus. An experimental study with comments on the intrinsic trigeminal connections.

The medullary projection from the mesencephalic trigeminal nucleus has been studied in cats where the wheat germ agglutinin-horseradish peroxidase complex has been used as a retrograde and anterograde tracer. All injections were made at the level of the caudal pole of the inferior olive and show that it is especially the lateral part of nucleus parvicellularis of the reticular formation which is the main area for termination of the fibres. In addition, it can not be excluded that the descending fibres also reach the medialmost part of the spinal trigeminal sensory nucleus (pars magnocellularis and the adjoining pars gelatinosa).

A projection from the raphe nuclei to the lateral reticular nucleus in the cat.

By means of retrograde transport of the wheat germ agglutinin-horseradish peroxidase complex afferent fibres to the lateral reticular nucleus from the raphe nuclei were demonstrated in the cat. The projection is bilateral and has its main origin in nucleus raphe pallidus and magnus, but there appear to be contributions also from nucleus raphe obscurus, dorsalis and linearis rostralis.