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The aim of the present study was to test for and characterize the organization of a direct projection from neurones of the trigeminal mesencephalic nucleus (Vme) to the cerebellum. WGA-HRP was used as a retrograde tracer following injections in the cerebellar cortex. The extent of each injection site within the sagittal zones was determined according to corticonuclear and olivocortical connections. Retrogradely labelled neurons were observed in the caudal part of the ipsilateral Vme only following vermal injection. The Vme projections reached exclusively the ipsilateral sagittal zone X in the anterior lobe, lobule VI and lobule IX. This identification was confirmed by anterograde labelling of mossy fibre terminals following a biocytin injection restricted to the Vme.

The amino acids GABA and glutamate (Glu) are thought to be the principal substances in the central nervous system responsible for neuronal inhibition and excitation. Their distributions among the different neurons in a defined pathway may thus be indicative of the contributions of the cells to pathway function. Examples of such neurons are those of the cerebellar nuclei which, while regulating output from the Purkinje cells of the cerebellar cortex, are also found to project back to the cerebellar cortex. Immunohistochemical experiments were done to identify GABA and glutamate (Glu) containing cells in the adult rat cerebellar nuclei. Consecutive semithin and serial vibratome sections were incubated with antisera raised in rabbit against GABA and Glu. In semithin sections, only small neurons were intensely GABA immunoreactive (GABA-IR) (31.7%), and the majority (80.5%) were Glu immunoreactive (Glu-IR) of different sizes. Consistent with Glu being a metabolic precursor for GABA, 75.4% of the GABA-IR population colocalized Glu. In vibratome sections GABA-IR neurons showed some local differences in number, whereas the Glu-IR were uniformly distributed in the three nuclei studied. Measured mean diameters for these neurons showed a distinct size difference for the GABA- and Glu-IR with little overlap. Cerebellar nuclei neurons projecting to the cortex (nucleocortical neurons, NCN) were identified by locally preinjecting the retrograde transported WGA-apoHRP-colloidal gold complex in the cerebellar cortex. Vibratome sections of these cerebellar were silver intensified for the retrograde tracer and double labeled for GABA and Glu. Of the total number of identified NCN, 8.7% were GABA-IR (10 animals) and 47.7% Glu-IR (5 animals). Many retrograde labeled NCN in the core of the thick sections were immunonegative for both amino acids due to poor antibody penetration, thus underestimating the proportions of cells containing GABA and Glu. The size distributions for the GABA-IR and Glu-IR NCN were similar to those measured in non-retrograde labeled nuclei in thick sections. The conclusions reached are that GABA-IR neurons of the cerebellar nuclei, including the NCN, use GABA as the presumed inhibitory neurotransmitter and that Glu-IR neurons may use Glu or another excitatory neurotransmitter.

Possible projections of serotonin (5-HT)-immunoreactive neurons in the nodose ganglia (NG) to the nucleus tractus solitarius (NTS) were investigated in the rat using a double labeling method combining retrograde transport and 5-HT immunohistochemistry. After injection of a complex of colloidal gold-apo-horseradish peroxidase into the medio-caudal and commissural parts of the NTS, most of the 5-HT-immunoreactive neurons were found to be labelled by the gold complex. The present study provides direct evidence for the existence, in the rat, of a serotonergic NG-NTS system. This system may be involved in the regulation of blood pressure and vigilance states.

The serotoninergic innervation of the inferior olivary complex of the rat was studied using a specific immunohistochemical technique. Fibers and varicosities positive for serotonin were present throughout the nucleus. The densest varicosities were found in the lateral portion of the dorsal accessory olive and the caudal portion of the medial accessory olive. The rostral medial accessory olive and the principal olive were sparsely populated with labeled elements. Ultrastructurally, labeled profiles were found in close opposition to small dendrites and to olivary cell bodies, but they did not display any synaptic specialization. Labeled perikaria were found in the periolivary regions, some of them located laterally to the olivary complex are responsible for the serotoninergic innervation of the dorsal accessory olive; some others located dorsally and medially in the nucleus raphe obscurus and raphe pallidus were responsible for the innervation of the medial accessory olive.

After a deposit of HRP crystals in one eye of larval and adult Pleurodeles and the PAP immunocytochemical procedure, the organization of the visual system was described at stages 36, 45; 52 and postmetamorphic (according to Gallien and Durocher, 1975) and in the adult. At hatching stage, the retinopreoptic and the accessory pathways differentiate, soon after the retino-thalamico-pretectal pathway appears. The tectum differentiates later and a true retinotectal bundle can be seen only after the end of metamorphosis. In the whole, the pattern of the normal system is very simple in Pleurodeles especially as regard the ipsilateral fibres which never reach the tectum. This rudimentary organization lead us to conclude that the Pleurodeles and the Protopterus visual patterns are very similar.