Caudal Magnocellular Nucleus Research Articles

Representation of waveform periodicity in the auditory midbrain of the bullfrog, Rana catesbeiana.

The period of complex signals is encoded in the bullfrog's eighth nerve by a synchrony code based on phase-locked responding. We examined how these arrays of phase-locked activity are represented in different subnuclei of the auditory midbrain, the torus semicircularis (TS). Recording sites in different areas of the TS differ in their ability to synchronize to the envelope of complex stimuli, and these differences in synchronous activity are related to response latency. Cells in the caudal principal nucleus (cell sparse zone) have longer latencies, and show little or no phase-locked activity, even in response to low modulation rates, while some cells in lateral areas of the TS (magnocellular nucleus, lateral part of principal nucleus) synchronize to rates as high as 90-100 Hz. At midlevels of the TS, there is a lateral-to-medial gradient of synchronization ability: cells located more laterally show better phaselocking than those located more medially. Pooled all-order interval histograms from short latency cells located in the lateral TS represent the waveform periodicity of a biologically relevant complex harmonic signal at different stimulus levels, and in a manner consistent with behavioral data from vocalizing male frogs. Long latency cells in the caudal parts of the TS (cell sparse zone, caudal magnocellular nucleus) code stimulus period by changes in spike rate, rather than by changes in synchronized activity. These data suggest that neural codes based on rate processing and time domain processing are represented in anatomically different areas of the TS. They further show that a populationbased analysis can increase the precision with which temporal features are represented in the central auditory system.

A new fixation procedure for study of the histaminergic neurons by immunoelectron microscopy using the direct antiserum against histamine.

A new perfusion protocol was developed to detect histamine-like immunoreactive neurons at the electron microscopic level. By stepwise perfusion of 1-ethyl-3(3-diamethylaminopropyl)-carbodiimide and paraformaldehyde solutions, the brain block could be cut with a vibratome and the immunoreactivity could be detected using the avidin-biotin-peroxidase-complex method. We used this method to study the ultrastructure and synaptic relations of the histaminergic neurons in the postmammillary caudal magnocellular nucleus of the rat hypothalamus. This method should also be useful for examination of histaminergic neurons in other tissues and the synaptic relations of histaminergic neurons with other neurotransmitter-containing neurons by double immunostaining.

Histaminergic neurons receive substance P-ergic inputs in the posterior hypothalamus of the rat

The synaptic connections between histaminergic neurons and substance P (SP) afferents in the caudal magnocellular nucleus (CM) of the hypothalamus were examined using an immunoelectron microscopic mirror method. SP-immunoreactive (SP-IR) terminals made synaptic contacts with the somata, somatic spines and dendrites of histidine decarboxylase immunoreactive (HDC-IR) neurons. This suggests that SP afferents exert monosynaptic influence on the central histaminergic neuronal system.

Neuropeptide Y afferents have synaptic interactions with histaminergic (histidine decarboxylase-immunoreactive) neurons in the rat brain.

The synaptic interaction was identified between histaminergic cells and neuropeptide Y (NPY)-immunoreactive axon terminals in the caudal magnocellular nucleus, by means of the immunoelectron microscopic mirror method. This observation suggests that NPY afferents exert monosynaptic influences upon the functioning of the central histaminergic neuronal system.

Development of histamine-immunoreactive neurons in the rat brain.

This study was undertaken to reveal the cellular stores of histamine in developing rat brain and to determine the stage of development during which the histamine-immunoreactive neurons can first be detected. Rats from embryonal day 12 to postnatal day 14 were studied. The brains were fixed in 4% 1-ethyl-3(3-dimethylaminopropyl)carbodiimide and standard immunofluorescence technique was used. The first histamine-immunoreactive neurons were seen on embryonic day 13 in the border of mesencephalon and metencephalon. On embryonic day 15 immunoreactive neurons were detected in ventral mesencephalon and rhombencephalon. In caudal, tuberal, and postmammillary caudal magnocellular nuclei histamine-immunoreactive neurons were first detected on embryonic day 20 while those in the hindbrain had disappeared. Histamine-immunoreactive nerve fibers were first detected on embryonic day 15 in rhombencephalon and mesencephalon and in some areas of diencephalon including the mammillary bodies and frontal cortex. On embryonic day 18 the number of immunoreactive nerve fibers in the hindbrain had decreased considerably, but the olfactory bulb, septal and hypothalamic area, and the cerebral cortex showed immunoreaction in fibers. The density of histamine-immunoreactive fiber networks increased until postnatal day 14 when an adultlike pattern of neurons and fibers had developed. Histamine-immunoreactive neurons are present in embryonal CNS and they develop extensive projections to various brain areas.

The histaminergic system in the guinea pig central nervous system: an immunocytochemical mapping study using an antiserum against histamine.

Using an antiserum against conjugated histamine we mapped the histaminergic somata and their fiber projection areas in carbodiimide-fixed guinea pig central nervous system. The neurons were large and they were found exclusively in the posterior hypothalamus, as in the rat, but in the guinea pig they were more numerous and distributed more widely in thin layer around the posterior mammillary nucleus, scattered between and within the medial mammillary nuclei, and in a dense cell cluster emerging from the caudal magnocellular nucleus and extending to the medial preoptic area. The density of histamine-immunopositive fibers was very high in the olfactory tubercle, diagonal band of Broca, nucleus accumbens, medial and cortical amygdaloid nuclei, periventricular and lateral basal hypothalamus, paraventricular thalamus, and in a region from the medial central gray to the locus coeruleus and the parabrachial nucleus. Dense fiber networks were found in the piriform and entorhinal cortex, septum, dentate gyrus, and subiculum, in most parts of amygdala, and in many areas of the hypothalamus, thalamus, substantia nigra, raphe nuclei, inferior olivary, solitary tract and medial vestibular nuclei, and neurohypophysis. Medium fiber density was observed in the internal layers of the olfactory bulb, anterior olfactory nuclei, neocortex, zone CA1 of hippocampus, and many midbrain and hindbrain regions. Low density was present in the outer layers of the olfactory bulb, other parts of hippocampus, the globus pallidus, most of the caudatus-putamen, the cerebellar cortex, and the dorsal horn of the spinal cord. The retina and most of the myelinated white matter had single or no histaminergic fibers. It may be concluded from the results that most fibers seem to follow a ventromedial route to the forebrain, reaching the amygdala ventral to the medial forebrain bundle, the hippocampus via subiculum, and the hindbrain structures via the medial central gray. As compared to the rat, the fiber projections in the guinea pig brain were denser, particularly in the hippocampus, thalamus, pons-medulla, and neurohypophysis. The fiber densities in various regions of the guinea pig brain are compared to histamine receptor densities and the possible functions of histamine are discussed.

Histaminergic neuron system and its function.

Histamine as a neurotransmitter or a neuroregulator in the mammalian brain has been postulated on the basis of biochemical, pharmacological and neurophysiological studies (1–5). We have demonstrated the histaminergic neuron system in rat brain immunocytochemically (6,7) using antibody raised against histidine decarboxylase (HDC), the sole enzyme responsible for histamine synthesis, as a marker (8). Histamine neurons were found to be concentrated in the magnocellular nuclei in the posterior hypothalamic area of rat brain, such as the tuberai magnocellular nucleus, caudal magnocellular nucleus and postmammillary caudal magnocellular nucleus, while their fibers were found in many areas, such as the hypothalamus, cerebral cortex, olfactory nuclei, basal ganglia, hippocampus, septum, cerebellum, amygdaloid complex, nucleus of the diagonal band, bed nucleus of the stria terminalis, central gray, auditory system, medial vestibular nucleus, facial nucleus, dorsal parabrachial nucleus, and nucleus of the solitary tract (7). Similar results were observed using antibodies against histamine itself (9–11).

Coexistence of galanin-like immunoreactivity with catecholamines, 5- hydroxytryptamine, GABA and neuropeptides in the rat CNS

The coexistence of galanin (GAL)-like immunoreactivity (LI) with markers for catecholamines, 5-hydroxytryptamine (5-HT), GABA, or some neuropeptides was mapped in the rat CNS by using adjacent sections, as well as by elution-restaining and double-labeling immunocytochemistry. Many instances of coexistence were observed, but there were also numerous GAL-positive cell body populations displaying distributions similar to those of these markers but without apparent coexistence. In the hypothalamic arcuate nucleus GAL-LI was found in a large proportion of tyrosine hydroxylase (TH)-positive cell bodies (A12 cells), both in the dorsomedial and ventrolateral subdivisions, with a higher number in the latter. GAL-LI coexisted in glutamic acid decarboxylase (GAD)-positive somata in the posterior aspects of the arcuate nucleus and at all rostrocaudal levels in fibers in the external layer of the median eminence. In the anterior hypothalamus, a large population of the cells of the parvocellular and magnocellular paraventricular nuclei contained both GAL-LI and vasopressin-LI. Moreover, somata containing both GAD- and GAL-LI were seen lateral to the mammillary recess in the tuberal and caudal magnocellular nuclei. Some of the neurons of the caudal group were shown to project to the occipital cortex using combined retrograde tracing and immunofluorescence. With regard to mesencephalic and medullary catecholamine neurons, GAL-LI coexisted in a large proportion of the noradrenergic locus coeruleus somata (A6 cell group) and in the A4 group dorsolateral to the fourth ventricle, as well as in the caudal parts of the A2 group in the dorsal vagal complex. However, in more rostral parts of the latter, especially in the medial subdivision of the solitary tract nucleus, a very large population of GAL-IR small cell bodies was seen intermingling with catecholamine neurons, but they did not contain TH-LI. Furthermore, GAL-IR cell bodies coextensive with, but not coexisting in, TH-IR somata were seen in the C1 (epinephrine) horea in the ventrolateral medulla at the level of area postrema and in the most rostral aspects of the C1 group. Finally, 5-HT-positive cell bodies of the mesencephalic and medullary raphe nuclei and a subpopulation of coarse 5-HT nerve fibers in the hippocampus co-contained GAL-LI. The present results demonstrate that a GAL-like peptide is present in many systems containing other neuroactive compounds, including dopamine, norepinephrine, 5-HT, GABA, and vasopressin.(ABSTRACT TRUNCATED AT 400 WORDS)

Histaminergic neurons in the rat brain: correlative immunocytochemistry, Golgi impregnation, and electron microscopy.

Histamine-containing neurons were visualized in Vibratome--sections of rat brain with the indirect peroxidase-antiperoxidase immunocytochemical method of Sternberger (Immunocytochemistry, 2nd edition, New York: John Wiley and Sons, pp. 1-354, '79) by utilizing a primary antibody directed against L-histidine decarboxylase (HDC). Cell bodies of HDC-immunoreactive neurons are located exclusively in the posterior hypothalamus: tuberal magnocellular nucleus (TM), caudal magnocellular nucleus (CM), and post-mammillary magnocellular nucleus (PCM). With the light microscope, all the HDC-immunoreactive neurons in CM and PCM and the majority of the HDC-immunoreactive neurons in TM appear to be large neurons, with a short, thick dendrite emerging from each pole of the long axis of the oval perikaryon and one or more, thinner, nonpolar primary dendrites. In the electron microscope, it can be seen that the immunoreaction product is diffusely dispersed in the cytoplasm. The ultrastructural features of all investigated (70) HDC-immunoreactive neurons in the three nuclei, independent of their light microscopic characteristics, are remarkably similar: large, unindented, pale nucleus; a high proportion of cytoplasm to nucleus (with the exception of the medium-sized HDC-immunoreactive neurons in TM); large, perinuclear array of Golgi apparatus; numerous mitochondria; endoplasmic reticulum fragmented into numerous small cisterns; thick initial portions of the primary dendritic trunks; few axosomatic synaptic contacts. Twenty-one Golgi-Kopsch-impregnated neurons taken from CM, PCM, and TM were embedded in epoxy resin, serially sectioned, and investigated in the electron microscope. The ultrastructural characteristics typical of HDC-immunoreactive neurons were observed in all three nuclei in neurons with large cell bodies tapering into two thick, sparsely spinous primary dendrites that subsequently dichotomize into very long (up to 100 microns), nontapering, aspinous secondary dendrites. In sections taken from the posterior hypothalamic area of rats prepared in a conventional way for electron microscopy, distinct populations of large cells can be observed in TM, CM, and PCM displaying the same set of ultrastructural characteristics as the HDC-immunoreactive neurons.

On the innervation of trigeminal mesencephalic primary afferent neurons by adenosine deaminase-containing projections from the hypothalamus in the rat

The localization and sources of adenosine deaminase-containing structures in the mesencephalic nucleus of the trigeminal nerve of the rat was studied using indirect immunofluorescence or immunoperoxidase immunohistochemical staining techniques for adenosine deaminase in combination with retrograde fluorescent tracing or lesion methods. The majority of large mesencephalic neurons were engulfed by a dense adenosine deaminase-immunoreactive plexus. Immunostaining was often punctate surrounding neuronal profiles or sometimes had the appearance of varicose fibers coursing over the neuronal surface. Occasionally, inununostained axons were found travelling towards and contacting mesencephalic neurons. Mesencephalic neuronal somas surrounded by immunofluorescence staining for adenosine deaminase were simultaneously labelled with fast blue after injections of this dye into the temporalis or masseter muscles of mastication. Injections of fast blue into the mesencephalic nucleus resulted in fast blue labelling of adenosine deaminase-immunoreactive neurons in the tuberal, caudal and postmammillary caudal magnocellular nuclei of the hypothalamus. Ablation of these hypothalamic nuclei caused a near total depletion of adenosine deaminase-immunostained fibers in the mesencephalic nucleus including those associated with mesencephalic neurons. It is concluded that adenosine deaminase-containing neurons in the posterior hypothalamus innervate mesencephalic primary sensory neurons, which are known to convey proprioceptive input to trigeminal motor nuclei controlling jaw muscles. The possibility is considered that the hypothalamus, via a direct action on these sensory neurons, may exert automatic control over jaw movements related to aggressive attack, defensive or feeding behavior. In addition, it appears that mesencephalic neurons may provide an ideal model system for electrophysiological investigations of the neurotransmitter(s) utilized by adenosine deaminase-containing hypothalamic projections.

The cytoarchitecture, histochemistry and projections of the tuberomammillary nucleus in the rat

The normal morphology, efferent projections and possible neurotransmitter content of neurons in the tuberomammillary nucleus (caudal magnocellular nuclei of Bleier et al.) [ Bleier, Cohn and Siggelkow (1979) In Anatomy of the Hypothalamus, Vol. 1, pp. 137–220] have been examined in the adult male rat. In Nissl-stained sections, the nucleus can be divided into a dorsomedial, ventral and diffuse part, each of which consists of large, darkly stained neurons cradling the mammillary body. The ventral part is by far the largest and consists of some 2500 neurons on each side of the brain. Immunohistochemical studies indicate that a majority of the large neurons in all three parts of the nucleus stain with antisera against glutamate decarboxylase and [Met]enkephalyl-Arg 6-Phe 7 heptapeptide and that a smaller subset of these neurons (about 10%) also stain with an antiserum against substance P. Single injections of retrogradely transported fluorescent tracers were made into 18 different sites in 86 animals and the results indicate that all three parts of the tuberomammillary nucleus on one side of the brain send fibers to or through various parts of the neocortex, hippocampal formation, amygdala, basal ganglia, thalamus, superior colliculus and cerebellum on both sides of the brain and that the projection neurons are not organized in a highly topographic way. Injections of two different fluorescent tracers in the same animal indicate that individual neurons in the nucleus may give rise to both ascending and descending projections, as well as projections to widely divergent parts of the forebrain. Together with previous results, this evidence suggests that the tuberomammillary nucleus has widespread projections to the numerous brain structures located in the forebrain and in the caudal medulla (it may not project to the spinal cord), and that its axons may release a mixture of neuroactive substances including γ-amino butyrate and several peptides. Although its functional significance remains to be investigated, morphological evidence suggests that the tuberomammillary nucleus may constitute one of a series of neurotransmitter-specific cell groups in the brainstem and basal forebrain with diffuse efferent projections that may be involved in the modulation of attention or behavioral state, rather than the processing of specific sensory or motor information.

Immunohistochemical evidence for the coexistence of histidine decarboxylase-like and glutamate decarboxylase-like immunoreactivities in nerve cells of the magnocellular nucleus of the posterior hypothalamus of rats.

Immunohistochemical staining of alternate consecutive sections revealed numerous histidine decarboxylase (L-histidine carboxy-lyase, EC 4.1.1.22)-like immunoreactive neurons that also contained glutamate decarboxylase (L-glutamate 1-carboxy-lyase, EC 4.1.1.15)-like immunoreactive structures in the tuberal magnocellular nucleus, the caudal magnocellular nucleus, and the postmammillary caudal magnocellular nucleus of the posterior hypothalamus of rats. Furthermore, in immunohistochemical double-staining procedures, almost all neurons in the magnocellular nuclei had both histidine decarboxylase-like and glutamate decarboxylase-like immunoreactivities. These results suggest the coexistence of histamine and gamma-aminobutyric acid in single neurons in these nuclei.

Cortical projections of monoamine oxidase-containing neurons from the posterior hypothalamus in the rat

Monoamine oxidase (MAO) histochemistry combined with retrograde horseradish peroxidase tracing of neurons demonstrated that MAO-containing neurons in the tuberal, caudal and postmamillary caudal magnocellular nuclei of the hypothalamus project to various cerebral cortices.

Fine structure of histaminergic neurons in the caudal magnocellular nucleus of the rat as demonstrated by immunocytochemistry using histidine decarboxylase as a marker.

The morphology of histamine-containing neurons in the caudal magnocellular nucleus was light and electron microscopically examined by means of peroxidase-antiperoxidase (PAP) immunocytochemistry with histidine decarboxylase (HDC) as a marker. HDC-like immunoreactive (HDCI) neurons had large (25-30 microns in diameter) perikarya from which two to four primary dendrites arose. The perikarya had a nearly round nucleus and well-developed Golgi apparatus in addition to a large number of mitochondria and rough endoplasmic reticulum. Immunoreactive endproducts were found diffusely throughout the perikarya, dendrites, and axons. HDCI neurons made synaptic contact with nonreactive axon terminals on the perikarya and dendrites. In addition, the HDCI neurons very frequently formed puncta adherentia with neuronal elements, either HDCI or nonreactive, or glial cells. Most of the HDCI axon terminals serially observed under electron microscopy did not exhibit typical synaptic contact in the caudal magnocellular nucleus. These findings suggest the nonsynaptic release of histamine in the caudal magnocellular nucleus.

5-HT immunoreactive hypothalamic neurons in rat and cat after 5-HTP administration

Using immunohistochemistry for serotonin (5-HT) after administration of 5-hydroxytryptophan (5-HTP), we have demonstrated the existence of a particular group of 5-HT accumulating neurons in the posterior hypothalamus of the rat and cat. This cluster of neurons, which we termed “5-HT immunoreactive (IR) posterior hypothalamic magnocellular neurons,” was characterized by: (1) diffuse 5-HT-IR staining; (2) rather large-sized cell bodies; (3) a marked activity of monoamine oxydase (MAO); (4) capacity of uptake and decar☐ylation of 5-HTP; (5) the absence of capacity of uptake and hydroxylation of tryptophan. These 5-HT-IR magnocellular neurons extended from the suprachiasmatic nucleus area to the caudal end of the hypothalamus. In the rat, the majority of these neurons was found in the tuberal and caudal magnocellular nuclei, whereas in the cat, it was found more widespread in the ventrolateral part of the posterior hypothalamus including the tuberomamillary nucleus. The characteristics of the 5-HT-IR magnocellular neurons which project directly both to the cerebral cortex and the pontine tegmentum are discussed in comparison with those of “APUD” cell series or “paraneurons,” as well as with those of other 5-HT-IR neurons demonstrated in the hypothalamus after administration of 5-HTP or tryptophan.

Immunohistochemistry of adenosine deaminase: implications for adenosine neurotransmission.

Immunohistochemical analysis of adenosine deaminase in rat brain revealed an extensive plexus of adenosine deaminase-containing neurons in the basal hypothalamus. These neurons converged on and were most numerous in three major centers, namely, the tuberal, caudal, and postmammillary caudal magnocellular nuclei. Most other brain regions were devoid of cells containing adenosine deaminase. Some adenosine deaminase-containing neurons were retrogradely labeled with the fluorescent dye fast blue when the dye was injected into the frontal cortex and striatum. Specific populations of neurons having high levels of adenosine deaminase may release adenosine as a neurotransmitter.

Histamine-containing neurons in the rat hypothalamus.

A specific antiserum against histamine was produced in rabbits, and an immunohistochemical study of histamine-containing cells was carried out in rat brain. The antiserum bound histamine in a standard radioimmunoassay and stained mast cells located in various rat and guinea pig tissues. Enterochromaffin-like cells in the stomach and neurons in the posterior hypothalamic area could be detected with this antiserum. The staining was highly specific and was not abolished by preabsorption with histidine, histidine-containing peptides, serotonin, or catecholamines, whereas preabsorption with histamine completely abolished the staining. Immunoglobulins of this antiserum purified by affinity chromatography stained the same cells as did the crude antiserum, whereas the serum fraction, which was not absorbed by histamine-affinity ligand, failed to stain any neuron. Histamine-immunoreactive neuronal cell bodies were found only in the hypothalamic and premammillary areas of colchicine-treated rats. The largest group of cells was seen in the caudal magnocellular nucleus and medially on the dorsal and ventral aspects of the ventral premammillary nucleus. Immunoreactive nerve fibers, but no cell bodies, were detected in other parts of the brain. Histamine-immunoreactive mast cells were found in the median eminence and pituitary gland. The results suggest that histamine-containing neurons are located only in a small area of the posterior hypothalamus, and these cells are probably the source of ascending and descending fibers detected in other brain areas.

Distribution of the histaminergic neuron system in the central nervous system of rats; a fluorescent immunohistochemical analysis with histidine decar☐ylase as a marker

The distribution of histidine decar☐ylase-like immunoreactivity (HDCI) in the rat central nervous system was studied by the indirect immunofluorescence technique. HDCI cell bodies were concentrated in the posterior hypothalamic area, such as in the tuberal magnocellular nucleus, caudal magnocellular nucleus, posterior hypothalamic nucleus and lateral hypothalamus just lateral to the fasciculus mammillothalamicus at the level of the posterior hypothalamic nucleus. Extensive networks of HDCI fibers of various densities were found in many areas of the brain; they were particularly dense in the hypothalamus but were also found in the following areas: rostrally in the cerebral cortex, olfactory nuclei, medial amygdaloid nucleus, n. tractus diagonalis, and bed nucleus of the stria terminals, and caudally in the central gray matter of the midbrain and pons, auditory system, n. vestibularis medialis, n. originis nervi facialis, n. parabrachialis, n. commissuralis, n. tractus solitarii, and n. raphe dorsalis.

Hypothalamic gamma-aminobutyric acid neurons project to the neocortex.

Three groups of gamma-aminobutyric acid--containing neurons were found in the mammillary region of the posterior hypothalamus. The groups correspond to the tuberal, caudal, and postmammillary caudal magnocellular nuclei. Many cells in these nuclei were retrogradely labeled with fast blue after the injection of this fluorescent dye into the neocortex. Immunohistochemical experiments showed that these same neurons also contained the gamma-aminobutyric acid-synthesizing enzyme glutamate decarboxylase. These results provide morphological evidence for a gamma-aminobutyric acid pathway arising in magnocellular neurons of the posterior hypothalamus and innervating the neocortex.

Descending projections to the inferior olive from the mesencephalon and superior colliculus in the cat. An autoradiographic study.

Descending projections from the mesencephalon and superior colliculus to the inferior olive were analyzed by an autoradiographic tracing method. Injections of tritium-labelled leucine were placed in regions which had previously been identified as sources of afferents to the olive. These were located adjacent to the central gray and extended from the rostral red nucleus to the posterior thalamus. Additional injections were made in the superior colliculus. Other injections were placed in the basal ganglia and thalamus. Injections restricted to one side of the central mesencephalon resulted in predominantly ipsilateral labelling of the olive. After injections in the caudo-medial parafascicular and subparafascicular nuclei and rostral nucleus of Darkschewitsch, deposits of grains were observed in the rostral pole of the medial accessory olive and adjacent ventral lamella of the principal olive. The medial accessory olive contained grains into its middle third. More caudal injections which involved the interstitial nucleus of Cajal as well as the nucleus of Darkschewitsch and rostral red nucleus resulted in the dense labeling of the entire principal olive (except the dorsal cap), the entire medial accessory olive (except subnucleus beta and the caudo-medial pole) and the caudo-dorsal accessory olive. Injections centered in the caudal magnocellular red nucleus and extending into the rostral parvocellular division labelled the dorsal lamella of the principal olive almost exclusively. When only the caudal part of the red nucleus was involved in the injection, the olive was entirely clear of grains. Minor contralateral distributions were observed in the dorsomedial cell column, the medial tip of the dorsal lamella and in the caudal medial accessory olive. The deep layers of the superior colliculus were found to project strongly to the contralateral medial accessory olive immediately beside subnucleus beta and weakly to the same area ipsilaterally. Four pathways were identified as contributing fibers to the olivary projections. These were the medial longitudinal fasciculus, the medial tegmental tract, the central tegmental tract and tectospinal or tectobulbar fibers. The rubrospinal tract did not contribute projections to the olive. Injections in the caudate nucleus, entopeduncular nucleus and ventral anterior and ventral lateral thalamic nuclei, did not result in any labeling in the olive.