Centrifugal innervation of goldfish retina from ganglion cells of the nervus terminalis

Cobaltous-lysine was applied to one optic nerve of normal goldfish in order to determine the source of the centrifugal innervation of the retina. Cobalt-filled cells were not observed in the optic tectum, pretectum, thal-amus, or hypothalamus. However, filled cells were observed outside the central nervous system either interspersed between the olfactory nerve fibers orrostrally along the ventromedial aspect of the olfactory bulbs. These cells appear to correspond to the ganglion cells of the nervus terminalis. The cells were located bilaterally and had dendrites that branched in close proximity to the cell body and axons that coursed caudally through the medial olfactory tract. The axons traveled in the ventral forebrain and entered the optic tracts. The axons also gave off fine branches that appeared to terminate in the vicinity of the anterior commissure and in the preoptic region. Application of cobaltous-lysine to a cut olfactory tract resulted in cobalt-filled fibers in the optic tracts, retinal optic fiber layer, and retinal ganglion cell layer. However, the precise terminations of these fibers within the retina could not be readily established. The results are discussed with respect to the plethora of sources of retinopetal cells observed by others in fish and with respect to the innervation of the retina by luteinizing hormone-releasing hormone axons.     

Chromophores in the extraretinal photoreceptor (pineal organ) of teleosts

We have demonstrated, using a highly sensitive high performance liquid chromatography technique, that the pineal organ (epiphysis cerebri) of the rainbow trout contains both 11-cis-retinal and11-cis-3-dehydroretinal. These compounds were assumed to be the chromophores of pineal photopigments, because they were isomerized to all- trans forms by light-adaptation. The quantity of chromophores in the dark-adapted pineal organ was extremely low (9.2–29.1 pmol/pineal), corresponding to approximately 1/300–1/1000 of that of the retina.     

A subpopulation of nervus terminalis neurons projects to the olfactory mucosa in Xenopus laevis

Biocytin application to the normal or zinc sulfate-treated nasal cavity of Xenopus laevis was used to trace retrogradely neurons associated with the terminal nerve (TN). Immunocytochemistry was conducted to identify the relationship of gonadotropin-releasing hormone-immunoreactive (GnRH-ir) TN neurons with biocytin-labeled neurons. Neurons that accumulated biocytin were located in olfactory nerve fascicles close to the olfactory mucosa lining the caudal, medial, and rostral walls of the principal cavity. GnRH-ir fibers were observed only in the olfactory nerve fascicle projecting to the rostral edge of the principal cavity. In addition, GnRH-ir fibers did not contact biocytin-labeled TN neurons. We hypothesize that these two classes of neurons represent separate components of the TN.     

LHRH-immunoreactive neurons in the pterygopalatine ganglia of voles: a component of the nervus terminalis?

Cranial tissue from adult and neonatal voles was examined with immunocytochemical techniques to determine the distribution of luteinizing hormone-releasing hormone-immunoreactive (LHRH-ir) neurons in extracerebral structures. The overall distribution of the LHRH-ir portion of the nervus terminalis in the nasal and extracranial cavities was comparable to that of other rodents. However, we observed a unique association of LHRH-ir neurons with the pterygopalatine ganglia of neonatal and adult voles. We also found LHRH-ir fibers in nasopalatine nerves, and trigeminal nerves and ganglia of neonatal voles. We speculate that these neurons may influence the autonomic control of the vascular pump in the vomeronasal organ.     

Corticotropin releasing factor-like immunoreactive neurons in the rat retina

Corticotropin releasing factor (CRF)-like immunoreactive neurons have been identified in the rat retina by immunohistochemical methods using antisera directed against ovine and rat CRF. CRF-like immunoreactivity was observed in both amacrine and ganglion cells which projected fine varicose processes to the inner plexiform layer of the retina. It is suggested that CRF may play a role in retinal function.     

Ontogenesis of neurons producing luteinizing hormone-releasing hormone (LHRH) in the nervus terminalis of the rat

Immunoreactive luteinizing hormone-releasing hormone (LHRH) was first detected at 15 days of gestation in ganglion cells associated with the peripheral, intracranial, and central parts of the nervus terminalis of the rat. LHRH was not detected in any other structure of the central nervous system at this age. In the 17-day-old fetal rat, 62% of the total LHRH-reactive neuronal population was found in ganglion cells of the nervus terminalis. At this same age, immunoreactive beta-luteinizing hormone (beta-LH) was first seen in gonadotropes of the anterior pituitary gland. At 19 days of gestation, 31% of the total number of LHRH-reactive neurons observed in the rat brain was found in the nervus terminalis, and immunoreactive processes were first seen in the organum vasculosum of the lamina terminalis and in the median eminence. Our data indicate that from 15 to 19 days of gestation the nervus terminalis is a principal source of LHRH in the fetal rat. Presence of the decapeptide in the nervus terminalis prior to appearance of beta-LH in the anterior pituitary suggests a possible role for LHRH in this system on maturation of the gonadotropes and differentiation of the brain-pituitary-gonadal axis.     

Projections of the olfactory bulb and nervus terminalis in the silver lamprey

The connections of the olfactory bulb were traced using horseradish peroxidase. A homologue of the medial olfactory tract in gnathostomes projects to the ipsilateral septal nucleus, preoptic area and, possibly, the rostral striatum. A homologue of the lateral olfactory tract projects to the ipsilateral lateral pallium, dorsal pallium and, possibly, the medial pallium, as well as to the posterior diencephalon. A component of the lateral olfactory tract decussates in the habenular and posterior commissures and distributes to the contralateral hemisphere and caudal diencephalon. A dorsal component of secondary olfactory fibers terminates, ipsilaterally, in a dorsomedially situated neuropil that has previously been interpreted as a single glomerulus of the olfactory bulb or as an accessory olfactory bulb, as well as in the contralateral olfactory bulb after decussation in the dorsal commissure. Afferents to the olfactory bulb arise from the ipsilateral dorsal pallium, lateral pallium, a cell-poor region adjacent to the preoptic area, and the midbrain tegmentum. The extent of the secondary olfactory projections in silver lampreys could be interpreted to support the phylogenetic hypothesis that all regions of the telencephalon received secondary olfactory projections in the earliest vertebrates, but this interpretation is not unequivocal, due to questions concerning the pallial homologues in lampreys and gnathostomes. Application of horseradish peroxidase to the olfactory epithelium revealed projections to the striatum, preoptic area, hypothalamus and posterior tuberculum that are comparable to projections of the nervus terminalis in other vertebrates.     

Orexin B immunoreactive fibers and terminals innervate the sensory and motor neurons of jaw-elevator muscles in the rat

The relationship between orexinergic terminals and the sensory and motor neurons of jaw-elevator muscles was examined by means of anti-orexin B (OXB) immunohistochemistry combined with horseradish peroxidase (HRP) retrograde tracing in the rat. HRP was initially injected into the jaw-elevator muscles; 48 h later the animals were sacrificed and fixed for HRP reaction and anti-OXB immunohistochemistry. OXB-like terminals were observed to distribute in the trigeminal mesencephalic nucleus (Vme) and the trigeminal motor nucleus (Vmo) where they closely contact the Vme neuronal somata and the Vmo neuronal somata and dendrites retrogradely labeled with HRP. The results of this study provide anatomical evidence of a direct OXB orexinergic innervation of the sensory and motor neurons controlling jaw-elevator muscles involved in mastication. Its functional significance related to the feeding behavior and bruxism is discussed.     

Bombesin immunoreactive neurons in the hypothalamic paraventricular nucleus innervate the dorsal vagal complex in the rat

Bombesin-like immunoreactivity has been localized within neuronal cell bodies of the hypothalamus and nerve terminals within the dorsal vagal complex. The possibility that the hypothalamus is a source for bombesin-like immunoreactive terminals within the dorsal vagal complex was examined using the combined retrograde tracing and immunohistochemical technique. After injections of retrograde tracer were made into the dorsal vagal complex, cells in the hypothalamus labeled with both retrograde tracer and bombesin immunoreactivity were localized in the parvocellular part of the paraventricular nucleus. In the paraventricular nucleus most of the vagal projecting bombesin immunoreactive neurons were located within the medial parvocellular subdivision. Approximately 30% of the bombesin immunoreactive neurons in this subnucleus projected to the dorsal vagal complex. The results suggest that the paraventricular hypothalamic nucleus is a major source of bombesin terminals within the dorsal vagal complex. This pathway may mediate some of the autonomic nervous system changes that are observed when bombesin is injected within the central nervous system. Additionally, this data adds to a growing amount of evidence supporting the role of bombesin as a peptide neurotransmitter.     

Effects of lesions of the optic nerve, optic tectum and nervus terminalis on rod precursor proliferation in the goldfish retina.

Teleost retinas grow throughout life by proliferation of neuroblasts at the retinal margin and dedicated rod precursors in the outer nuclear layer. Mechanisms regulating this proliferation are largely unknown. Previous investigators observed that rod precursor replication, as detected by incorporation of radioactive thymidine into cells of the outer nuclear layer, is enhanced after optic nerve crush. We attempted to determine whether this was due to severing of the retinopetal (nervus terminalis, n.t.) or retinofugal (retinal ganglion cell) axons in the optic nerve of the goldfish, Carassius auratus. In the first series of experiments, we ablated unilaterally the optic nerve, olfactory bulb (containing n.t. ganglia), or optic tectum (containing retinal ganglion cell axons and n.t. collaterals). Rod precursor proliferation increased dramatically in both retinas as soon as 5 days after surgery; in addition, the numbers of dividing cells were greater in the ipsilateral retina 10-15 days after optic nerve crush or tectal ablation and in the contralateral retina 20-25 days after olfactory bulb ablation. These observations are not accounted for by the known projections of retinal ganglion cells, but are consistent with the projections of the n.t. In the second series of experiments, n.t. projections to the brain and retina were severed bilaterally 7-8 weeks before the unilateral optic nerve crush or hemitectal ablation. Rod precursor proliferation increased as before, but the quantities of dividing cells were always equal in both retinas. We conclude that the n.t. may modulate rod proliferation locally and that injury to (some) brain regions may cause release of mitogens that affect rod precursors in both retinas.     

Antibodies against retinal photoreceptor-specific proteins reveal axonal projections from the photosensory pineal organ in teleosts

With the aid of specific antisera to the retinal proteins S-antigen and alpha-transducin and to the rhodopsin apoprotein opsin, we have labeled various cell populations in the pineal organ, parapineal organ, habenular nucleus, and subcommissural organ in two teleost species: the rainbow trout and the European minnow. Although these proteins are associated with photoreceptor functions, not only photoreceptor cells but also the majority of parenchymal cells in the pineal organ were immunoreactive. Immunoreactive cells with dendrite- and axonlike processes were observed also in the parapineal organ and the habenular nucleus. Furthermore, S-antigen-immunoreactive, long, axonal processes were observed in the pineal organ and could be traced from the pineal organ to the habenular nucleus and to the pretectal area. In the light of recent HRP electron microscopical and immunocytochemical studies we propose (1) that not only the classical pineal photoreceptor cells of poikilothermic vertebrates but also other types of CSF-contacting neurons may be the phylogenetic ancestors of mammalian pinealocytes, and (2) a close interrelationship between the pineal organ and the limbic system, effectuated by the direct projections from pineal photoreceptors/CSF-contacting neurons/pinealocytes to the habenular nucleus, and by displaced "pinealocytelike" elements scattered in the habenular nucleus.     

Central projections of the nervus terminalis in four species of amphibians

The central projections of the nervus terminalis were investigated in two anuran and two urodele species by means of horseradish peroxidase injections into one nasal cavity. In anurans, the nervus terminalis projects to the medial septum, to the preoptic nucleus, to the nucleus of the anterior commissure and to the hypothalamus. In addition to these structures, the dorsal thalamus, the infundibulum and the mesencephalic tegmentum are innervated in urodeles. The structure containing the highest density of terminals in the amphibians investigated is the hypothalamus. In one anuran and one urodele species, the contralateral hypothalamus is primarily innervated, whereas in the other two species the majority of fibers remain ipsilateral. A comparison with other vertebrates shows that the terminalis system in urodeles has the greatest diversity of connections. Anurans, in contrast, lack some connections that are present in urodeles and fishes. These findings have implications for a possible relation of the nervus terminalis to an aquatic habitat.     

Morphology and distribution of neurons immunoreactive for substance P in the turtle retina

Immunocytochemical staining procedures with the HRP-complexed antibody to substance P have been carried out on the turtle retina. Examination by light microscopy of wholemount retinas has allowed us to evaluate the morphology and distribution of the substance P immunoreactive cell types. Two amacrine cell types and two or more ganglion cell types are stained in our hands. Type A amacrines are tri-stratified wide-field amacrines. They have their major dendrites in S1 and S3 of the inner plexiform layer and they emit fine dendrites from the major dendrites that end in varicose boutons in S5 on and around cell bodies in the ganglion cell layer. Some of the dendrites in S1 radiate out in axon-like fashion for 1 mm across the retina. The type B amacrine cells are small to medium-field in dendritic extent. They have smaller cell bodies than type A and a single or, at most, two primary dendrites that pass directly to S3 before branching profusely into an intricate net-like dendritic field. The ganglion cells that are stained with substance P antibodies appear to be of several types but their exact morphologies are in doubt because only portions of their major dendrites are stained. Substance P immunoreactive axons are clearly seen to project from the cell bodies to the optic nerve head and axons are stained in the optic nerve itself. The substance P-stained ganglion cells occur in an irregular distribution that reaches a peak density in an elongated band parallel to and 1 mm below the visual streak. The type B amacrine cells reach a maximum density in the visual streak and are distributed in a highly regular mosaic decreasing in density in elliptical isodensity contours from the visual streak. In contrast the type A amacrine cells are rare or absent in the streak, being located in an irregular mosaic in peripheral retina.     

Central neural connections of the pineal organ and retina in the teleost Gasterosteus aculeatus L

The relations of the central neural connections of the pineal organ to those of the retinae of the lateral eyes were investigated in the three-spined stickleback, Gasterosteus aculeatus L. (Teleostei), by anterograde and retrograde transport of horseradish peroxidase (HRP). HRP was applied to the crushed pineal stalk and/or injected into the left or the right eye. Both pineal and retinal efferents project to area praetectalis, dorsal and ventral thalamic areas, and dorsal tegmentum. The most notable overlapping occurs in nucleus commissurae posterioris of area praetectalis. Pineal efferents also innervate the habenular nuclei and dorsal hypothalamus, while retinal efferents innervate rostral hypothalamus, ventrolateral thalamus, and tectum opticum. A small number of retinofugal axons recross and innervate the ipsilateral nucleus anterioris periventricularis and area praetectalis. After intraocular HRP injections, labeled perikarya were located both in retinofugal terminal areas and in areas not receiving direct retinal input, such as the telencephalic nucleus olfactoretinalis, deep tectal layers, and an area rostroventral to nucleus dorsolateralis thalami. No neurons afferent to the pineal organ were demonstrated. The close association of pineal efferents with retinofugal and possible retinopetal elements is in accordance with the view that both systems are potential neural mediators of photoperiodic events in the teleostean circadian system.     

Central projections of the nervus terminalis in lampreys, lungfishes, and bichirs

Central projections of the nervus terminalis were investigated in a cyclostome (Lampetra planeri), in a sarcopterygian (Protopterus dolloi), and in an actinopterygian fish (Polypterus palmas), following the injection of horseradish peroxidase into the olfactory epithelium. Despite differences in forebrain morphology (inversion versus eversion of the hemispheres), projections of the terminal nerve are similar in the species investigated. The nervus terminalis courses through the subpallium (septum) and mainly innervates periventricular nuclei in the telencephalon and diencephalon. In lampreys, the majority of labeled fibers terminate in the hypothalamus, while in bony fishes the main projection is to periventricular nuclei of the anterior commissure. The course of the nervus terminalis through the dorsomedial telencephalon in lungfishes supports the interpretation that this part of the brain constitutes the septum, and not a pallial structure. Nervus terminalis projections are compared with those in teleosts and in amphibians. The presumed lack of gonadotropin-releasing hormone in the nervus terminalis of lampreys is discussed.     

Light responses and morphology of bNOS‐immunoreactive neurons in the mouse retina

Nitric oxide (NO), produced by NO synthase (NOS), modulates the function of all retinal neurons and ocular blood vessels and participates in the pathogenesis of ocular diseases. To further understand the regulation of ocular NO release, we systematically studied the morphology, topography, and light responses of NOS‐containing amacrine cells (NOACs) in dark‐adapted mouse retina. Immunohistological staining for neuronal NOS (bNOS), combined with retrograde labeling of ganglion cells (GCs) with Neurobiotin (NB, a gap junction permeable dye) and Lucifer yellow (LY, a less permeable dye), was used to identify NOACs. The light responses of ACs were recorded under whole‐cell voltage clamp conditions and cell morphology was examined with a confocal microscope. We found that in dark‐adapted conditions bNOS‐immunoreactivity (IR) was present primarily in the inner nuclear layer and the ganglion cell layer. bNOS‐IR somas were negative for LY, thus they were identified as ACs; nearly 6% of the cells were labeled by NB but not by LY, indicating that they were dye‐coupled with GCs. Three morphological subtypes of NOACs (NI, NII, and displaced) were identified. The cell density, intercellular distance, and the distribution of NOACs were studied in whole retinas. Light evoked depolarizing highly sensitive ON‐OFF responses in NI cells and less sensitive OFF responses in NII cells. Frequent (1–2 Hz) or abrupt change of light intensity evoked larger peak responses. The possibility for light to modify NO release from NOACs is discussed. J. Comp. Neurol. 518:2456–2474, 2010. © 2010 Wiley‐Liss, Inc.     

Morphology and mosaics of VIP-like immunoreactive neurons in the retina of the rhesus monkey.

Vasoactive intestinal peptide (VIP) is a 28-amino acid peptide that has been demonstrated to reside in cells ( = VIP+ cells) of the retinae of various vertebrate species. In an attempt to study the morphology and distribution of VIP+ cells in the retina of the rhesus monkey in more detail, we subjected VIP+ cells observed in cryostat sections or wholemounts rhesus monkey retinae to a quantitative analysis. VIP+ cells were found to reside in the innermost row of the inner nuclear layer (INL) and in the ganglion cell layer (GCL) in similar numbers (estimate: 50 cells/mm2 at 6-10 mm eccentricity each) and only on rare occasions (12% of all VIP+ cells) in varying positions within the inner plexiform layer (IPL). Somata of VIP+ cells were circular and had a mean diameter of 9.1 microns. They gave rise to 1-3 main dendrites, which were usually oriented toward the IPL. Main dendrites ramified widely into thin fibers (dendritic field diameter less than = 1 mm), carrying varicose swellings. The fibers that contributed to one and the same plexus of VIP+ fibers preferred the middle third of the IPL, independent of the positions of the parent somata. A quantitative analysis of nearest-neighbour distances in the retinal wholemount preparation suggested that VIP+ cells in the GCL and in the INL might be distributed according to 2 independent mosaics. A comparison with Golgi-stained material leads to the tentative equation of VIP+ cells with the "spiny" A12 amacrine cell of Mariani ('90). Whereas the low density and large dendritic field size of VIP+ cells might suggest a more widespread function, the varicose dendritic morphology seems to be more compatible with functionally independent dendritic subunits mediating localized effects.     

Melatonin synthesis: arylalkylamine N-acetyltransferases in trout retina and pineal organ are different

Serotonin N-acetyltransferase (AANAT) is the first enzyme in the conversion of serotonin to melatonin. Changes in AANAT activity determine the daily rhythm in melatonin secretion. Two AANAT genes have been identified in the pike, pAANAT-1 and pAANAT-2, expressed in the retina and in the pineal, respectively. The genes preferentially expressed in these tissues encode proteins with distinctly different kinetic characteristics. Like the pike, trout retina primarily expresses the AANAT-1 gene and trout pineal primarily expresses the AANAT-2 gene. Here we show that the kinetic characteristics of AANAT in these tissues differ as in pike. These differences include optimal temperature for activity (pineal: 12 degrees C; retina: 25 degrees C) and relative affinity for indoleethylamines compared to phenylethylamines. In addition, retinal AANAT exhibited substrate inhibition, which was not seen with pineal AANAT. The kinetic differences between AANAT-1 and AANAT-2 appear to be defining characteristics of these gene subfamilies, and are not species specific.     

Persistence of the nervus terminalis in adult bats: a morphological and phylogenetical approach

The presence of the terminalis system in adult bats is demonstrated by light microscopical investigation of several species of Microchiroptera. In late embryonic and fetal stages of the mouse-eared bat (Myotis myotis) the compact central terminalis ganglion gradually differentiates into a three-dimensional network of cord-like ganglia and fiber bundles. Rostrally the terminalis system is in immediate contact with the medial-most fila olfactoria; caudally terminalis rootlets attach near the border between the olfactory bulb and the septum of the brain. With respect to the findings presented here it seems likely that all mammals develop a terminalis system in early ontogenesis and retain it until the adult stage. However, considerable differences concerning the number of persisting neurons may be found among some mammalian orders.     

Amygdaloid input to transiently tyrosine hydroxylase immunoreactive neurons in the bed nucleus of the stria terminalis of the rat

Several studies have reported transient expression of tyrosine hydroxylase in a subpopulation of neurons in the bed nucleus of stria terminalis of preadolescent rats. The tyrosine hydroxylase immunoreactive (TH) neurons, which are of small to medium size and often display a typical bipolar configuration, are confined to the intermediate part of the lateral bed nucleus. By the use of a combination of experimental tracer techniques and immunocytochemical methods, we have demonstrated that these neurons receive a significant number of amygdaloid afferents, which establish mostly symmetric synaptic contacts on the cell bodies and sparsely spined dendritic shafts of the TH neurons. TH neurons also receive a small number of tyrosine hydroxylase-positive terminals of unspecified origin.