The neurofilament architecture of the rat suprachiasmatic nucleus

The neurofilament architecture within the suprachiasmatic nucleus of the rat was analyzed immunocytochemically using neurofilament monoclonal antibodies. The topographic distribution of neurofilament containing structures was restricted mainly to the ventral and caudal part of the suprachiasmatic nucleus, coinciding with the entrance area of the retino-suprachiasmatic fibres of this nucleus. Within the nucleus itself an axonal organization was present. The axons were grouped, forming clusters. These clusters existed of a core of myelinated axons surrounded by unmyelinated axons. The myelinated/unmyelinated axon ratio could reach 1:25. Within the nucleus the myelinated axons extended upwards to the middle part of the suprachiasmatic nucleus, where the fibers of the axon clusters fanned out.     

The motor nuclei of the glossopharyngeal-vagal and the accessorius nerves in the rat

Retrograde cobalt labeling was performed by incubating the rootlets of cranial nerves IX, X and XI, or the central stumps of the same nerves, in a cobaltic lysine complex solution, and the distribution of efferent neurons sending their axons into these nerves was investigated in serial sections of the medulla and the cervical spinal cord in young rats. The following neuron groups were identified. The inferior salivatory nucleus lies in the dorsal part of the tegmentum at the rostral part of facial nucleus. It consists of a group of medium-sized and a group of small neurons. Their axons make a hair-pin loop at the midline and join the glossopharyngeal nerve. The dorsal motor nucleus of the vagus situates in the dorsomedial part of the tegmentum. Its rostral tip coincides with the first appearance of sensory fibres of the glossopharyngeal nerve, the caudal end extends into the pyramidal decussation. The constituting cells have globular or fusiform perikarya and they are the smallest known efferent neurons. The ambiguous nucleus is in the ventrolateral part of the tegmentum. The rostral tip lies dorsal to the facial nucleus, and the caudal tip extends to the level of the pyramidal decussation. The rostral one third of the ambiguous nucleus is composed of tightly-packed medium sized neurons, while larger neurons are arranged more diffusely in the caudal two thirds. The long dendrites are predominantly oriented in the dorsoventral direction. The dorsally-oriented axons take a ventral bend anywhere between the ambiguous nucleus and dorsal motor nucleus of the vagus. The motoneurons of the accessorius nerve are arranged in a medial, a lateral and a weak ventral cell column. The medial column begins at the caudal aspect of the pyramidal decussation and terminates in C2 spinal cord segment. The lateral and ventral columns begin in C2 segment and extend into C6 segment. The neurons have large polygonal perikarya and characteristic cross-shaped dendritic arborizations. The axons follow a dorsally-arched pathway between the ventral and dorsal horns. The accessorius motoneurons have no positional relation to any of the vagal efferent neurons. It is concluded that the topography and neuronal morphology of accessorius motoneurons do not warrant the designation of a bulbar accessorius nucleus and a bulbar accessorius nerve.     

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.     

Primary optic projections in the rabbit. Study using the horseradish peroxidase anterograde labeling technic

The primary visual pathways, in particular those to the lateral geniculate body, of 11 albino and 7 pigmented rabbits, were studied using the method of anterograde labelling with horseradish peroxidase following injection of the tracer into the vitreous body of one eye. A heavy projection to the contralateral dorsal lateral geniculate nucleus was seen in all animals. In both albino and pigmented animals a region devoid of label was present in the medial part of the alpha sector of the nucleus. This region corresponded to a compact, oval or wedge-shaped field of terminal label in the ipsilateral nucleus, which was much heavier in pigmented than in albino rabbits. In the ventral lateral geniculate nucleus, contralateral retinal input was almost entirely confined to the caudal half of the lateral sector of the nucleus, where two laminae of dense terminal label, separated by a less densely labelled area, were oriented parallel to one another and to the optic tract. This bilaminar distribution of retinal afferents to the ventral lateral geniculate nucleus has not been described in previous studies. The ipsilateral projection was to the dorsal part of the lateral sector and was most prominent in pigmented animals. The "intergeniculate leaflet" received a prominent contralateral input in all animals, and a clear ipsilateral input in pigmented animals, which overlapped with the contralateral input. Projections to other primary visual centres (pretectal nuclei, superior colliculus, nuclei of the accessory optic tract) are also described.     

Projection of the neurons of the substantia nigra and ventral field of the midbrain tegmen in the cat to the putamen

By means of retrograde axonal transport of the horseradish peroxidase and fluorochromes in the cat, it has been stated that neurons of all the parts of the substantia nigra (SN) make projections to the putamen. These projections are organized in such a way that the rostral part of the putamen gets the projected fibers from less number of the SN parts than the caudal part. To the caudal part of this formation all parts of the SN are projected, and in the equal degree to its dorsal and ventral segments. Projections to the rostral part are sent only from two parts of the SN--compact and dorsal. To the dorsal segment of this part only axons from a small amount of the nigral neurons are sent. A small amount of neurons of the tegmental ventral field give projections to the ventral segments along the whole rostrocaudal extent of the putamen. Convergence of the SN neuronal axons in the formations of the striated body has been determined, as well as overlapping of the terminal fields in the putamen from the projective neurons of the nigral various parts. Besides in different parts of the SN discrepancy has been revealed in the neuronal populations, labelled with different stainings, that contain cells, marked with two markers, injected into the nucleus caudatus and into the putamen.     

Localization of galanin-like immunoreactive structures in the brain of the Senegalese sole, Solea senegalensis

The distribution of galanin-like immunoreactive structures was studied in the brain of the Senegalese sole, Solea senegalensis, using immunohistochemical methods. Periventricular immunoreactive cell bodies were observed in the rostral pole of the preoptic recess, within the pars parvocellularis of the nucleus preopticus parvocellularis. Another galanin-immunoreactive cell population was observed more caudal in the ventromedial hypothalamus, along the medial evaginations of the lateral recess. These cells appear within the cytoarchitectonic limits of the nucleus recessus lateralis pars ventralis. We found an extensive presence of galanin-immunoreactive fibres throughout the entire brain, although the most massive network of fibres was observed in the caudal olfactory bulbs, ventral telencephalon, preoptic area and around diencephalic ventricular recesses. Also, the hypophysis, ventricular mesencephalic area, median reticular formation and viscerosensory rhombencephalon displayed important plexuses of galanin-immunoreactive axons.The widespread distribution of these immunoreactive structures in the brain and pituitary of the Senegalese sole suggests an important role for galanin in neuroendocrine regulation of brain and adenohypophyseal functions.     

Functional implications of the projections of neurons from individual labellar sensillum of Drosophila melanogaster as revealed by neuronal-marker horseradish peroxidase

Summary Earlier studies using Golgi silver impregnations from the labellar sensilla of adult Drosophila melanogaster revealed seven types of sensory axons projecting into the suboesophageal ganglion of the brain. These sensory terminals were designated as coiled fibres (type-I), shrubby fibres (type-II), ipsilateral ventral fibres (type-III), ipsilateral dorsal fibres (type-IV), contralateral ventral fibres (type-V), contralateral dorsal fibres (type-VI), and central fibres (type-VII). The present study identifies the projections of sensory neurons present in a single labellar taste-sensillum, using the neuronal marker horseradish peroxidase (HRP). Although the taste sensillum in question has five neurons, in a given experiment only one or at the most two neurons are labelled. The type of neuron labelled was usually specific to the stimulant solute (sucrose, sodium chloride or potassium chloride) present in the HRP solution. Although type-II fibres get labelled most of the time, irrespective of the stimulant present in HRP solution, type-IV fibres are labelled when attractants (0.1 M sucrose or 0.1 M sodium chloride) are used as stimulants in HRP solution. Type-VI fibres are labelled when the stimulant is 0.1 M potassium chloride, a repellent. HRP dissolved in distilled water revealed type-I coiled fibres. Besides revealing projections of sensillar neurons to the brain the present technique also inferred their possible function. Incubation of whole-brain tissue with 0.04% 3,3-diaminobenzidine tetrahydrochloride in presence of 0.06% hydrogen peroxide suggested that the glomerular organization is also present in the taste-sensory region as it is in olfactory neuropile.     

Central representation of the anterior lateral line nerve in dwarf catfish]

The central distribution of the anterior lateral line nerve (ramus) (NLLa) were studied in the pygmean sheat-fish (Ictalurus nebulosus) by means of the method of Nauta--Gygax. NLLa, as well as posterior lateral line nerve (NLLp), projects topographically on the nucleus medialis of the acoustic-lateral area. Within the nucleus medialis NLLa can be traces in the rostro-lateral parts. A majority of NLLa fibres can be followed ipsilaterally, although a definite contralateral contribution also exists. It is concluded that NLLa, representing the electro- and mechanoreceptors of the head region, distributes predominantly within the rostral part of the medial nucleus, while NLLp, representing the trunk receptors, distributes in the caudal half of the medial nucleus. There is some overlapping within the middle part of the nucleus.     

An immunohistochemical and chromatographic analysis of the distribution and processing of proneuropeptide Y in the rat suprachiasmatic nucleus

The suprachiasmatic nucleus (SCN) regulates a number of circadian rhythms in mammals. A neuropeptide Y (NPY)-containing pathway from the intergeniculate leaflet of the lateral geniculate to the SCN is considered to carry information of the environmental light-dark cycle. Antisera directed against NPY, Cys-NPY(32-36)amide or the C-terminal extended peptide of proNPY(68-97) (CPON) and avidin-biotin immunohistochemistry were used to define the precise distribution of NPYergic nerve fibers in the SCN, and to compare the location of the various fragments of proNPY in these nerves. Gel chromatography and specific radioimmunoassays were applied to quantify the efficiency of the amidation of NPY, and to study the size of peptides demonstrating NPY- and NPYamide-immunoreactivity in anterior hypothalamic extracts. NPY-, NPYamide-, and CPON-immunoreactive nerve fibers exhibited apparently the same distribution and morphology in the SCN. Immunoreactive fibers were preferentially located in the ventral part of the SCN, but along the rostrocaudal axis of the nucleus, the density and the precise distribution of immunoreactive elements changed. From the rostral third of the SCN to the middle third, the number of immunoreactive fibers increased and their distribution extended in a dorsal and lateral direction. In the caudal part of the SCN, the number of immunoreactive elements decreased and the innervation spread to an even more dorsolateral location. Dorsal aspects of the rostral SCN contained a moderate number of fibers, whereas the dorsomedial quadrant of the caudal 2/3 of the SCN was almost devoid of immunoreactivity.(ABSTRACT TRUNCATED AT 250 WORDS)     

Electron microscopic autoradiography of serotonin uptake in the ganglia of the fresh-water mussel (Anodonta cygneak L.).

After in vitro incubation, the uptake of labelled serotonin was investigated by electron microscopic autoradiography in the ganglia of fresh-water mussel (Anodonta cygnea L.) The labelled serotonin was primarily taken up by the axons of the neurons. The silver grains could always be localized over axons containing eccentric dense-core vesicles with a diameter of 100--200 nm. The results suggest (a) the possibility of the electron microscopic identification of serotonin-containing neurons, and (b) the direct role of the eccentric dense-core vesicles in the storage of serotonin.     

Substance P in the suprachiasmatic nucleus of the rat: an immunohistochemical and in situ hybridization study

Using a biotin-streptavidin-horseradish peroxidase (HRP) immunohistochemical technique the distribution of substance P-immunoreactive neuronal elements was investigated in the rat suprachiasmatic nucleus (SCN). Substance P-immunoreactive nerve fibres and varicosities were distributed throughout the suprachiasmatic nucleus, with the largest accumulation in its ventral part. Because this location overlaps with the innervation of retinal afferents, the distribution and density of substance P-immunoreactive fibres in bilaterally enucleated rats were compared to normal rats. The density of substance P-immunoreactive fibres and nerve terminals in the ventral part of the suprachiasmatic nuclei was reduced in the rats with bilateral destruction of the optic nerves, whereas the density of fibres and nerve terminals in the dorsal part as well as other retinal target areas in the thalamus and mesencephalon was unaffected. In rats pretreated with an intraventricular injection of colchicine several substance P-immunoreactive perikarya were identified in the suprachiasmatic nucleus. The immunoreactive neurons, measuring 9.7 m±1.1 m in diameter, were frequently observed in the central core of the nucleus and to a lesser extent in the dorsomedial and ventrolateral subparts. Using in situ hybridization histochemistry pre-protachykinin-A mRNA was found in the same part of the SCN indicating that synthesis of substance P takes place in SCN neurons. Using a double immunohistochemical approach applying diaminobenzidine and benzidinedihydrochloride as chromagens substance P-, vasoactive intestinal peptide (VIP)-, and vasopressin/neurophysin-immunoreactivities were identified in the same brain section. The substance P-immunoreactive perikarya constituted a separate population of SCN neurons, which were not vasopressin-, neurophysin- or VIP-immunoreactive. Taken together, these observations show that substance P is contained in the retinohypothalamic pathway and within a group of SCN cell bodies, indiating that substance P may play a role in the generation and entrainment of circadian rhythmicity.     

The vestibular nuclei in the domestic hen (Gallus domesticus). VII.--Afferents from the spinal cord

Lesions of different parts of the spinal cord at different levels in the hen have been made and the resulting degeneration in the vestibular complex has been studied in silver impregnated sections. Spinovestibular fibres originate from cervical as well as lumbosacral levels of the cord and run in the dorsal part of the lateral funiculus. The spinovestibular fibres from all levels of the spinal cord terminate ipsilaterally in the nucleus Deiters ventralis, the nucleus Deiters dorsalis, the medial nucleus and rostrally in the descending nucleus. The spinovestibular fibres terminating in the above nuclei are few in number while spinovestibular fibres terminating bilaterally in the caudal part of the descending nucleus are much more abundant. In a few cases HRP injections in the vestibular complex resulted in labelled cells in upper cervical segments of the spinal cord localized in lamina VII. The findings are discussed in the light of data concerning the spinovestibular pathway in mammals.     

Localization of heart-innervating sympathetic neurons

Localization, amount, form of the bodies and maximal diameter of horseradish peroxidase (HP)-labelled neurons in the right stellate ganglion (SG) in the cat spinal cord have been investigated. HP application has been performed on the central parts of the SG connective branch with vagus nerve, or with the caudal cardiac nerve. In the neurons HP has been revealed after Straus or Mesulam method. In the SG, regardless the HP application place, the labelled neurons arrange in the zone, adjoining the place, where the caudal cardiac nerve and the connective branch get to the vagus nerve. In the spinal cord, when HP is applied on the connective branch, the labelled neurons are revealed in the lateral horns of the TI-TVI segments. The amount of the labelled neurons decreases in the rostro-caudal direction. Their greatest amount is revealed in the TI-TIII segments. When HP is applied on the central part of the caudal cardiac nerve, a small amount of the labelled neurons has been found in TI-TIII segments of the spinal cord only in one experiment. Thus, in the connective branch of the SG with the vagus nerve much more amount of the preganglionar fibers run than in the caudal cardiac nerve.     

The suprachiasmatic nucleus in the sheep: retinal projections and cytoarchitectural organization

The retinal innervation, cytoarchitectural, and immunohistochemical organization of the suprachiasmatic nucleus (SCN) was studied in the domestic sheep. The SCN is a large elongated nucleus extending rostrocaudally for roughly 3 mm in the hypothalamus. The morphology is unusual in that the rostral part of the nucleus extends out of the main mass of the hypothalamus onto the dorsal aspect of the optic chiasm. Following intraocular injection of wheat-germ agglutininhorseradish peroxidase or tritiated amino acids, anterograde label is distributed throughout the SCN. Retinal innervation of the SCN is bilaterally symmetric or predominantly ipsilateral. Quantitative image analysis demonstrates that, although the amount of autoradiographic label is greatest in the ventral and central parts of the nucleus, density varies progressively between different regions. In addition to the SCN, retinal fibers are also seen in the medial preoptic area, the anterior and lateral hypothalamic areas, the dorsomedial hypothalamus, the retrochiasmatic area, and the basal telencephalon. Whereas the SCN can be identified using several techniques, complete delineation of the nucleus requires combined tract tracing, cytoarchitectural, and histochemical criteria. Compared with the surrounding hypothalamic regions, the SCN contains smaller, more densely packed neurons, and is largely devoid of myelinated fibers. Cell soma sizes are smaller in the ventral SCN than in the dorsal or lateral parts, but an obvious regional transition is lacking. Using Nissl, myelin, acetylcholinesterase, and cytochrome oxidase staining, the SCN can be clearly distinguished in the rostral and medial regions, but is less differentiated toward the caudal pole. Immunohistochemical demonstration of several neuropeptides shows that the neurochemical organization of the sheep SCN is heterogeneous, but that it lacks a distinct compartmental organization. Populations of different neuropeptide-containing cells are found throughout the nucleus, although perikarya positive for vasoactive intestinal polypeptide and fibers labeled for methionine-enkephalin are predominant ventrally; neurophysine-immunoreactive cells are more prominent in the dorsal region and toward the caudal pole. The results suggest that the intrinsic organization of the sheep SCN is characterized by gradual regional transitions between different zones.     

A well defined spinocerebellar system in the weakly electric teleost fish Gnathonemus petersii. A tracing and immuno-histochemical study.

Long ascending fiber systems were investigated in the spinal cord of a teleost fish, Gnathonemus petersii. Concomitant results of Fink-Heimer degeneration tracing as well as CaBP28K immunohistochemical labelling demonstrate the existence of a well defined direct pathway from the very lowest spinal level to the caudal lobe of the cerebellum. HRP retrograde labelling shows that this pathway originates in a cellular column located in the most ventral part of the lateral column next to the lateral extremity of the ventral horn. From each spinal segment, the large axons of these cells gather and form a strip shaped tract at the periphery of the lateral column immediately dorsal to the cell column from which they originate. The spinal course of these fibers is ipsilateral; they give off a large number of collaterals to the lateral reticular nucleus. Bypassing the trigeminal motor nucleus, the lateral column tract courses dorsally to the paratrigeminal command associated nucleus between the lateral lemniscus and the nucleus preeminentialis and with a ventro-dorsally oriented large loop, turns in the caudal direction and penetrates into the cerebellar caudal lobe. Running caudally in the dorsal granular layer of the caudal lobe, it shifts more and more medially and crosses the midline whilst decussating with the contralateral tract on the dorsal margin of the molecular layer of the caudal lobe. Finally, the tract splits off and terminates throughout the granular layer of the caudal lobe. The main characteristics of this pathway are similar to those of the ventral spinocerebellar tract of higher vertebrates; it conveys information from all spinal levels directly to the contralateral cerebellum. However, it does not seem to receive direct synaptic input from the periphery, since projection of the dorsal root fibers appears to be limited to the dorsal ipsilateral half of the spinal cord. The appearance of such a pathway in a teleost fish is probably related to the existence of a well developed proprioceptive system in this species.     

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.     

The early development of retinal ganglion cells with uncrossed axons in the mouse: retinal position and axonal course

The carbocyanine dye, DiI, has been used to study the retinal origin of the uncrossed retinofugal component of the mouse and to show the course taken by these fibres through the optic nerve and chiasm during development. Optic axons first arrive at the chiasm at embryonic day 13 (E13) but do not cross the midline until E14. After this stage, fibres taking an uncrossed course can be selectively labelled by unilateral tract implants of DiI. The earliest ipsilaterally projecting ganglion cells are located in the dorsal central retina. The first sign of the adult pattern of distribution of ganglion cells with uncrossed axons located mainly in the ventrotemporal retina is seen on embryonic day 16.5, thus showing that the adult line of decussation forms early in development. A small number of labelled cells continue to be found in nasal and dorsal retina at all later stages. At early stages (E14-15), retrogradely labelled uncrossed fibres are found in virtually all fascicles of the developing nerve, intermingling with crossed axons throughout the length of the nerve. At later stages of development (E16-17), although uncrossed fibres pass predominantly within the temporal part of the stalk, they remain intermingled with crossed axons. A significant number of uncrossed axons also lie within the nasal part of the optic stalk. The position of uncrossed fibres throughout the nerve in the later developmental stages is comparable to that seen in the adult rodent (Baker and Jeffery, 1989). The distribution of uncrossed axons thus indicates that positional cues are not sufficient to account for the choice made by axons when they reach the optic chiasm.     

Cross-midline interactions between mouse commissural hindbrain axons contribute to their efficient decussation

Information from both sides of the brain is integrated by axons that project across the midline of the central nervous system via numerous commissures present at all axial levels. Despite the accumulated experimental evidence, questions remain regarding the formation of commissures in the presence of strong repulsive signals in the ventral midline. Studies from invertebrates suggest that interaction at the midline between homologous axons of specific decussating neurons contributes to efficient midline crossing, but such evidence is lacking in vertebrate systems. We performed experiments to determine whether commissural axons of the caudal region of the hindbrain interact with their contralateral counterparts at the ventral midline and to evaluate the relevance of this reciprocal interaction. Double anterograde axon labeling with lipophilic tracers revealed close apposition between growth cones of contralateral pioneer decussating axons at the midline. Later, we detected fasciculation between contralateral axons that is maintained even after they have crossed the midline. Blocking axon projections unilaterally with a solid mechanical barrier decreased dramatically the midline crossing of the equivalent population from the contralateral side. Decussation was also blocked by a unilateral barrier permeable to diffusible molecules but not by an axon-permeable barrier. These results suggest that in the caudal region of the hindbrain, midline crossing is facilitated by interactions between decussating contralateral axon partners.     

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.     

Nucleus isthmi of the frog: structure and tecto-isthmic projection.

The morphology of neurons in the isthmic nucleus was studied with the Golgi technique. Most of the neurons have thick dendrites covered with lamelliform dendritic processes. The tecto-isthmic projection was investigated with the Fink--Heimer technique after partial tectal lesions. The anteromedial part of the tectum projects on the dorsal and anterior part, the caudomedial tectal region on the dorsal and posterior part of the nucleus. The posterolateral tectal area projects on the anterolateral part of the nucleus, and the axons originating in the anterolateral tectal quadrant terminate in its ventral and caudal part.