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.     

Demonstration of a sparse direct projection from the interstitial nucleus of Cajal to the cerebellum in the cat

In the cat the possible contribution of cerebellar afferents from the accessory oculomotor nuclei was investigated by means of retrograde transport of the wheat germ agglutinin-horseradish peroxidase complex. A sparse bilateral projection from the interstitial nucleus of Cajal was demonstrated. The fibres reach cerebellar cortical as well as nuclear regions. No retrogradely labelled cells were found in the nucleus of Darkschewitsch and the nucleus of the posterior commissure.     

The hypothalamo-cerebellar projection in the rat: origin and transmitter.

Hypothalamic neurons projecting to cerebellum were identified by retrograde tracing with wheat germ agglutinin-horseradish peroxidase (WGA-HRP) in the rat. Selective D-[3H]aspartate labelling was used to investigate whether any of these connections may use excitatory amino acids as transmitters. The WGA-HRP experiments revealed that the hypothalamo-cerebellar fibers have their main origins in the lateral, dorsal and posterior hypothalamic areas, and the tubero-mammillary nucleus, while smaller numbers of cells were observed in tuber cinereum, the anterior hypothalamic area, and the periventricular and paraventricular nuclei. After injections of D-[3H]aspartate into the cerebellar cortex, intense labelling of the olivocerebellar climbing fiber system was observed, but hypothalamic cells were not retrogradely labelled with this selective tracer. The absence of D-[3H]aspartate labelling indicates that hypothalamo-cerebellar neurons lack specific uptake mechanisms for excitatory amino acids, but it does not entirely preclude the possibility that some of these hypothalamic neurons may use such transmitters. Many cerebellar projecting cells were located in the tubero-mammillary nucleus, which is known to contain histaminergic and GABAergic neurons, and it was concluded that part of the hypothalamo-cerebellar pathways may use histamine and/or GABA as transmitters. The transmitter remains unknown for other parts of the hypothalamo-cerebellar pathways.     

Origins of nerve fibers containing nitric oxide synthase in the rat celiac-superior mesenteric ganglion

The origin of nitric oxide synthase-containing nerve fibers in rat celiac-superior mesenteric ganglion was examined using retrograde tracing techniques combined with the immunofluorescence method. Fluoro-Gold was injected into the celiac-superior mesenteric ganglion. Neuronal cell bodies retrogradely labeled with Fluoro-Gold in the thoracic spinal cord, the dorsal root ganglia at the thoracic level, the nodose ganglion, and the intestine from the duodenum to the proximal colon were examined for nitric oxide synthase immunoreactivity. About 60% of sympathetic preganglionic neurons in the intermediolateral nucleus projecting to the celiac-superior mesenteric ganglion were immunoreactive for nitric oxide synthase, as were approximately 27% of nodose ganglion neurons and about 65% of dorsal root ganglion neurons projecting to the cceliac-superior mesenteric ganglion. Neurons projecting to the celiac-superior mesenteric ganglion were found in the myenteric plexus of the small and large intestine. In the proximal colon, about 23% of such neurons were immunoreactive for nitric oxide synthase. However, in the small intestine, no immunoreactivity was found in these neurons.     

The neurons of origin of non-cortical afferent connections to the oculomotor nucleus. A retrograde labeling study in the rabbit.

The cellular origin of the brainstem projections to the oculomotor nucleus in the rabbit has been investigated by using free (HRP) and lectin-conjugated horseradish peroxidase (WGA-HRP). Following injections of these tracers into the somatic oculomotor nucleus (OMC), retrogradely labeled cells have been observed in numerous brainstem structures. In particular, bilateral labeling has been found in the four main subdivisions of the vestibular complex, predominantly in the superior and medial vestibular nuclei and the interstitial nucleus of Cajal, while ipsilateral labeling was found in the rostral interstitial nucleus of the medial longitudinal fascicle (Ri-MLF), the Darkschewitsch and the praepositus nuclei. Neurons labeled only contralaterally have been identified in the following structures: mesencephalic reticular formation dorsolateral to the red nucleus, abducens internuclear neurons, group Y, several areas of the lateral and medial regions of the pontine and medullary reticular formation, ventral region of the lateral cerebellar nucleus and caudal anterior interpositus nucleus. This study provides also information regarding differential projections of some centers to rostral and caudal portions of the OMC. Thus, the rostral one-third appears to receive predominant afferents from the superior and medial vestibular nuclei, while the caudal two-thirds receive afferents from all the four vestibular nuclei. Finally, the group Y sends afferents to the middle and caudal, but not to the rostral OMC.     

电刺激猫中脑中央灰质发音区的反应及该区的传入联系

刺激猫中脑中央灰质发音区可引起动物发音、情绪反应以及心律异常、血压升高等变化。同时,本研究还揭示,分别在中脑中央灰质嘴侧和尾侧发音区注入辣根过氧化物酶后,其逆行标记神经元分布相类似;但是,中脑中央灰质发音区和非发音区的传入联系则互不相同。     

Chemically defined collateral projections from the pons to the central nucleus of the amygdala and hypothalamic paraventricular nucleus in the rat

Triple fluorescence labelling was employed to reveal the distribution of chemically identified neurons within the pontine laterodorsal tegmental nucleus and dorsal raphe nucleus which supply branching collateral input to the central nucleus of the amygdala and hypothalamic paraventricular nucleus. The chemical identity of neurons in the laterodorsal tegmental nucleus was revealed by immunocytochemical detection of choline-acetyltransferase or substance P; in the dorsal raphe nucleus, the chemical content of the neurons was revealed with antibody recognizing serotonin. The projections were defined by injections of two retrograde tracers, rhodamine-and fluorescein-labelled latex microspheres, in the central nucleus of the amygdala and paraventricular nucleus, respectively. Neurons projecting to both the central nucleus of the amygdala and the paraventricular nucleus were distributed primarily within the caudal extensions of the laterodorsal tegmental nucleus and dorsal raphe nucleus. Approximately 11% and 7% of the labelled cells in the laterodorsal tegmental nucleus and dorsal raphe nucleus projected via branching collaterals to the paraventricular nucleus and central nucleus of the amygdala. About half of these neurons in the laterodorsal tegmental nucleus were cholinergic, and one-third were substance-P-ergic; in the dorsal raphe nucleus, approximately half of the neurons containing both retrograde tracers were serotonergic. These results indicate that pontine neurons may simultaneously transmit signals to the central nucleus of the amygdala and paraventricular nucleus and that several different neuroactive substances are found in the neurons participating in these pathways. This coordinated signalling may lead to synchronized responses of the central nucleus of the amygdala and paraventricular nucleus for the maintenance of homeostasis. Interactions between different neuroactive substances at the target site may serve to modulate the responses of individual neurons.     

Analysis of the efferent projections of the lateral geniculate nucleus with special reference to the innervation of the subcommissural organ and related areas

In order to define central neurons projecting to the subcommissural organ (SCO) and to related areas in the postero-medial diencephalon, Phaseolus vulgaris-leucoagglutinin (PHA-L) was injected into the lateral geniculate nucleus of the rat. PHA-L-labelled neurons send axonal processes medially through the posterior thalamic nuclei and the posterior commissure to the other hemisphere. Branches of fibres originating from this projection form a plexus of nerve terminals in the underlying precommissural nucleus and in the nucleus of the posterior commissure. A small number of PHA-L-immunoreactive nerve fibres penetrate from the precommissural nucleus into the lateral part of the SCO. A few labelled fibres penetrate directly from the posterior commissure into the medial part of the caudal SCO. Most of the PHA-L-immunoreactive fibres occur in the hypendymal layer, although a few terminate near the ependymal cells of the organ. Many labelled fibres are found in the ventricular ependyma adjacent to the SCO, some fibres lying close to the ventricular lumen. These results were obtained only if the tracer was delivered into the intergeniculate leaflet of the lateral geniculate nucleus (IGL). The IGL innervates both the suprachiasmatic nucleus and the pineal organ; the connections between the IGL and the midline structures, including the SCO, suggest that these areas are influenced by the circadian system.     

Brainstem neurons projecting to the ampullae of anterior,lateral, and posterior semicircular canals in the guinea pig

Neurons projecting to the ampullae of anterior, lateral, and posterior semicircular canals were identified in the guinea pig brainstem using horseradish peroxidase labeling techniques. Two groups of neurons forming bilateral connections were found, one located dorsally and the other ventrally to facial nerve trajectories. The dorsal group of vestibular efferent neurons projecting to all three canals was detected in the subependymal granular layer of the floor of the 4th ventricle lateral to the facial nerve genu and in the abducent nerve nucleus. Efferent neurons belonging to the ventral group were unevenly distributed through different areas of the parvocellularis nucleus and the rostral portion of the pontine caudal reticular nucleus. The morphological characteristics and distribution pattern of these cells are taken as confirmation of their heterogeneity of neuronal and functional organization in the vestibular efferent system of semicircular canal ampullae.A. A. Bogomolets Institute of Physiology, Academy of Sciences of the Ukrainian SSR, Kiev. Translated from Neirofiziologiya, Vol. 20, No. 4, 1988, pp. 526–532.     

Observations on the projection from the perihypoglossal nuclei onto the cerebellum in the macaque monkey

The occurrence and distribution of retrogradely labeled cells in the perihypoglossal nuclei of the monkey were mapped after injections of horseradish peroxidase in various cerebellar cortical regions. In general the findings are in accord with those made in the cat. The flocculus receives a heavy bilateral projection from the nucleus prepositus, particularly from its caudoventral part, and from the nucleus of Roller. There is an apparently scanty projection from the nucleus intercalatus. The uvula receives a rather similar projection, but in the prepositus the cells projecting to the uvula are on the whole situated more dorsally and rostrally than those supplying the flocculus. The projection to lobules VII-VIII is distinct. More scanty projections have been found to the paramedian lobule and the anterior lobe. The different but partially overlapping sites of origin in the prepositus of fibers to the flocculus and uvula indicate the presence of a topical pattern within the perihypoglosso-cerebellar projection, as in the cat (34). In the monkey the two regions of origin appear to coincide with two particular cell collections in the prepositus (12). Both small and middle sized cells project to the cerebellum, as they do in the cat (9, 48). The nucleus supragenualis nervi facialis in the macaque is morphologically different from the corresponding nucleus in most other mammalian species (12), but it contains labeled cells after injections in the flocculus, uvula and other cerebellar regions. A considerable number of cells in the abducent nucleus are labeled after injections in the flocculus and the posterior vermis.     

Ontogeny of afferents to the fetal rat cerebellum.

Afferents to the fetal rat cerebellum have been studied in fixed tissue with the fluorescent tracer, 1,1'-dioctadecyl-3,3,3',3'tetramethylindocarbocyanine perchlorate (DiI). The dye was applied to the cerebellar anlage at ages from embryonic day (E) 12 to birth (P0). Central processes of vestibular ganglion cells were found to be the first identifiable afferents to the cerebellum, being present at least by E13 and perhaps as early as E12. Ipsilateral spinocerebellar fibres may be labelled from E15, vestibular nuclei (both ipsi- and contralateral) also from E15, while contralateral inferior olivary nuclei could not be retrogradely labelled until E17. Trigeminocerebellar neurons in the interpolaris subnucleus of the nucleus of the trigeminal spinal tract and neurons of the lateral reticular nucleus were not labelled until E22 and P0, respectively. Finally, contralateral pontine nuclei were retrogradely labelled from the cerebellum after birth.     

Distribution of parasympathetic motoneurones in the oculomotor complex innervating the ciliary ganglion in the marmoset (Callithrix jacchus)

Parasympathetic motoneurones in the oculomotor complex which innervate the ciliary ganglion were identified using the horseradish peroxidase (HRP) retrograde axonal tracer technique. The ciliary ganglion was located behind the eye by a lateral orbital approach and injected with HRP pellets mounted on the tips of microelectrodes. Most of the labelled cells were distributed throughout the whole Edinger-Westphal nucleus (EW). Outside the EW, only a small number of labelled cells were found, and most of these were located in the median zone ventral to the EW. There was no evidence of a division of the EW into rostral and posterior subnuclei, nor for separate cell populations in the EW and the anterior median nucleus (AM), respectively. At rostral levels labelled cells were confined to the EW with no overlap into the AM. In contrast to most previous studies no labelled cells were found in the AM. The possible physiological functions of the EW and the ventral components of the EW are discussed.     

Afferent connections of the cat lateral vestibular nucleus

Location within the brain of HP-labeled neurons (origins of projections to the lateral vestibular nucleus) was investigated by iontophoretic injection of this enzyme. Bilateral projections to the following midbrain structures were revealed: the field of Forel, interstitial nuclei of Cajal, oculomotor nerve nuclei, and the red nucleus — to all parts of the lateral vestibular nucleus. Bilateral projections were also shown from more caudally located structures, viz. the superior, medial and inferior (descending) vestibular nuclei, Y groups of the vestibular nuclear complex, facial nucleus and hypoglossi, nucleus prepositus nervi hypoglossi and caudal nuclei of the trigeminal tract; ipsilateral projections from crus IIa of lobulus ansiformus of the cerebellar hemisphere; contralateral projections from the bulbar lateral reticular nucleus and Deiter's nucleus. A tonic organization pattern of afferent inputs from a number of brainstem formations to the dorsal and ventral lateral vestibular nucleus is revealed and trajectories of HP-labeled fiber systems projecting to Deiter's nucleus described.L. A. Orbeli Institute of Physiology, Academy of Sciences of the Armenian SSR, Erevan. Translated from Neirofiziologiya, Vol. 20, No. 4, pp. 494–503, July–August, 1988.     

The relationship between MI and SMA afferents and cerebellar and pallidal efferents in the macaque monkey

The purpose of the present study was to determine the interrelationship between the thalamic afferents arising from the cerebellum (Cb) and the internal segment of the globus pallidus (GPi) with the neurons projecting to the primary motor cortex (MI) and to the supplementary motor area (SMA). We combined fluorescent retrograde tracers with a double anterograde labeling technique. Multiple injections of a combination of Diamidino Yellow and Fast Blue were made into either the MI or SMA hand/arm representation as determined by intracortical microstimulation. In the same animal, biotinylated dextran amine was injected into the GPi and horseradish peroxidase conjugated to wheat germ agglutinin was injected into the contralateral cerebellar nuclei. The results revealed that the cerebellar and pallidal thalamic territories are largely separate. The ventral anterior nucleus (VA) and the ventral lateral nucleus pars oralis (VLo) contained a greater density of pallidal labeling while a greater density of cerebellar label was observed more caudally in the ventral posterior lateral nucleus pars oralis (VPLo) as well as in nucleus X (X). Moreover, we observed that the greatest coincidence of retrograde cell labeling was within the pallidal thalamic territory following the SMA injections and within the cerebellar thalamic territory following the MI injections. However, interdigitating foci of pallidal and cerebellar label were also observed particularly in the ventral lateral nucleus pars oralis (VLo) and the ventral lateral nucleus pars caudalis (VLc). In both VLo and VLc, we additionally observed coincidence between the cerebellar labeling and SMA projection neurons as well as between pallidal labeling and MI projection neurons. These data suggest that while MI primarily receives inputs originating from Cb and SMA primarily receives inputs originating from GPi, it also appears that MI and SMA receive secondary afferents arising from GPi and Cb, respectively.     

Localization and morphometric analysis of masticatory muscle afferent neurons in the nucleus of the mesencephalic root of the trigeminal nerve in the cat

Injections of horseradish peroxidase (HRP) were made into the ipsilateral temporal muscle and contralateral masseter muscle of 10 cats in order to identify and characterize neurons in the nucleus of the mesencephalic root of the trigeminal nerve that innervate muscle receptors in the orofacial periphery. Neurons labelled by HRP injections and unlabelled cells from 5 control cats were measured with a computer-based image analyzer, and their position was mapped on a stereotaxic graph. Cells that innervate the masseter and temporal muscles were identified throughout the rostrocaudal extent of the nucleus. There was no indication of a somatotopic pattern nor of a specific segregation within the nucleus for cells innervating muscle receptors. The nucleus contained small, rounded unipolar neurons located primarily in the dorsal border of the periaqueductal gray (PAG) matter in the rostral part of the nucleus and larger oval unipolar neurons which were scattered throughout the nucleus, but were predominant in the pontine portion of the nucleus. HRP injections labelled both large and small cells, as well as occasional multipolar cells. The last-mentioned tended to be located in the lateral margins of the PAG. The mean geometric values obtained for the control group were: area 552.7 microns2 perimeter 110.3 microns; maximum diameter 36.0 microns. and diameter of an equivalent circle 26.1 microns. The mean values of the labelled neurons were: area 606.6 microns2; perimeter 100.1 microns; maximum diameter 36.0 microns, and diameter of an equivalent circle 27.2 microns.     

Identification of tuberculo-ventral neurons in the polymorphic layer of the rat dorsal cochlear nucleus

The tuberculo-ventral tract represents a short nervous circuit within the auditory cochlear nuclei. Tuberculo-ventral neurons of the dorsal cochlear nucleus send isofrequency inhibitory inputs to bushy cells of the ventral cochlear nucleus. Injection of wheat germ agglutinin conjugated to horseradish peroxidase into the rat ventral cochlear nucleus, labelled tuberculo-ventral neurons retrogradely in the deep polymorphic layer of the ipsilateral dorsal cochlear nucleus. Five to 20% of the perimeter of these cells was covered by synaptic boutons, most of which contained flat and pleomorphic vesicles. These boutons contained glycine and sometimes GABA. Occasional small axo-somatic boutons contained round vesicles and were immunonegative for both glycine and GABA. This study shows that the synaptic profile of tuberculo-ventral neurons is different from that of other medium-size glycinergic neurons within the polymorphic layer or more superficial regions of the dorsal cochlear nucleus like cartwheel neurons. In fact the latter mostly receive boutons that contain pleomorphic vesicles.     

Morphology of neurons and axon terminals associated with descending and ascending pathways of the lateral forebrain bundle in Rana esculenta

Summary The cells of origin of afferent and efferent pathways of the lateral forebrain bundle were studied with the aid of the cobalt-filling technique. Ascending afferents originated from the lateral thalamic nucleus, central thalamic nucleus, posterior tuberculum and the cerebellar nucleus. They terminated in the anterior entopeduncular nucleus, amygdala and the striatum. Telencephalic projection neurons, which are related to the lateral forebrain bundle, were located mainly in the ventral striatum and the anterior entopeduncular nucleus, but were not so numerous in the dorsal striatum. Irrespective of their location, most of the neurons projecting axons into the lateral forebrain bundle had piriform or pyramidal perikarya. Long apical dendrites usually arborized in a narrow space, whereas widely arborizing secondary dendrites originated from short dendritic trunks. The other neurons that contributed to the lateral forebrain bundle were fusiform or multipolar cells. Striatal efferents terminated in the pretectal area and in the anterodorsal, anteroventral and posteroventral tegmental nuclei.     

Cellular localization of 3H-oestradiol in the hypothalamus

Summary The hypothalamus of male and female rats, given 0.3 g/100 g body weight of 6.7-³H-oestradiol-17 and killed 1 hour after the injection, was examined by autoradiography in order to 1) localize the areas and the cells involved in the uptake of the hormone, and 2) study the intracellular localization of the labelled material.Only nerve cells contained radioactive material while glial and ependymal cells were not significantly labelled. In the anterior hypothalamus, labelled nerve cells were concentrated in areas corresponding to nucleus preopticus medialis and nucleus preopticus, pars suprachiasmatica. The nucleus supraopticus was unlabelled. In the medial basal hypothalamus, neurons corresponding to the nucleus arcuatus and the lateral part of the nucleus ventromedialis showed marked labelling. No significant labelling was observed in the nucleus paraventricularis, pars magnocellularis.Although the individual nerve cells varied in their extent of labelling, the major proportion of the silver grains were consistently concentrated over the nuclei. Castration was not found to influence the results. The findings were essentially the same in male and female rats and appear to suggest that oestradiol exerts a direct effect on nerve cells in certain hypothalamic areas.This work was supported by grants from the Norwegian Cancer Society, Nordisk Insulinfond and Anders Jahres Fond. The skilful assistance of Miss Helga Friedl and Mrs. Jane Larsen is gratefully acknowledged.     

The nucleus raphe dorsalis of the rat and its projection upon the caudatoputamen. A combined cytoarchitectonic, immunohistochemical and retrograde transport study

The nucleus raphe dorsalis of the albino rat has been studied in the following three ways: (1) the cell mass was subjected to a detailed cytoarchitectonic analysis, based upon Nissl-stained material; (2) serotonin--as well as the noradrenaline--immunoreactive neurons present in the area of the nucleus raphe dorsalis were plotted; (3) following large injections of the fluorescent dye propidium iodide into the caudatoputamen complex, the cells in the nucleus raphe dorsalis projecting to this complex were labeled and subsequently stained with an antibody against serotonin. Cytoarchitectonic analysis showed that three cell types are present within the confines of the nucleus raphe dorsalis: small, medium and large. Moreover, differences in concentrations of cell bodies made it possible to subdivide the nucleus raphe dorsalis into four regions. Immunohistochemical analysis showed that the borders of the serotoninergic cell groups B6 and B7 of DAHLSTROM and FUXE do not coincide with those of the nucleus raphe dorsalis. Serotonin-immunoreactive perikarya in the nucleus raphe dorsalis were categorized as medium and large neurons; noradrenaline-immunoreactive neurons in the nucleus raphe dorsalis do all belong to the category--large neurons. With the combined use of immunofluorescence and fluorescent retrograde tracing, it was found that the projection from the nucleus raphe dorsalis to the caudatoputamen complex originates from serotoninergic as well as non-serotoninergic cells, both of which can be categorized as being medium-sized neurons. The data presented in this paper provides a guide for further studies of afferent and efferent connections of the nucleus raphe dorsalis and for electrophysiological experiments on its constituent neurons.     

Sources of thalamic afferents projecting to the suprasylvian gyrus of the black sea porpoise

Neurons sending fibers to different loci of the suprasylvian gyrus (SSG) of the porpoise(Phocaena phocaena) cortex were located in the thalamus by retrograde horseradish peroxidase transport and fluorescent tracing techniques. Horseradish peroxidase injection into the anterior section of the suprasylvian gyrus led to retrograde labelling of neurons in the lateral portion of the ventrobasal complex of nuclei and the ventroposteroinferior nucleus. A group of labelled cells was found in the ventral section of the main medial geniculate nucleus. Injecting bisbenzimide into different loci of the medial suprasylvian gyrus also led to retrograde labelling of neurons belonging to the ventral division of the main medial geniculate nucleus. Somewhat lower numbers of labelled cells were found in the inferior nucleus of the pulvinar. Small groups of labelled neurons were also found in the lateral nucleus of the pulvinar, the medioventral nucleus of the medial geniculate body, and the posterior complex of nuclei. A similar distribution of labelled cells was also observed after injecting bisbenzimide into the more caudal portion of the gyrus, although the location of labelled cells in the ventral division of the main medial geniculate nucleus and the lower pulvinar nucleus were shifted in a lateral direction.A. N. Severtsov Institute of Animal Evolutionary Moprhology and Ecology, Academy of Sciences of the USSR, Moscow. National University, Singapore. Translated from Neirofiziologiya, Vol. 21, No. 4, pp. 529–539, July–August, 1989.