Distribution of somatostatin in the rat brain: Telencephalon and diencephalon

The distribution of somatostatin (SRIF) was examined using the unlabeled antibody enzyme method of immunocytochemistry on thick 30-50 microns Vibratome sections. The greatest population of SRIF-neurons was observed along the ventricular wall in the preoptic area and anterior hypothalamus. Dense accumulations of fibers were observed in the suprachiasmatic, ventromedial and arcuate nuclei, the internal and external zone of the median eminence, the organum vasculosum of the lamina terminalis and the subfornical organ. Extrahypothalamic sites of SRIF-containing neurons and fibers were also observed throughout the telencephalon. The widespread distribution of SRIF is consistent with radioimmunoassay data and suggests a diverse physiological role for somatostatin.     

Capsaicin-induced neuronal degeneration: silver impregnation of cell bodies, axons, and terminals in the central nervous system of the adult rat

Capsaicin is a neurotoxic substance valued in neurobiological research because of its ability to selectively damage small unmyelinated primary sensory neurons. Previous work has indicated that systemic capsaicin administration causes permanent neuronal degeneration in neonatal rats, but evidence that capsaicin has a similar effect in adults is equivocal and incomplete. Therefore, we used silver impregnation, a method that labels degenerating neurons, to examine the central nervous system of adult rats after systemic capsaicin treatment. Adult rats were injected with a single intraperitoneal dose of capsaicin (50 or 90 mg/kg) or vehicle solution and killed 6, 12, 18, 24, 48, 96, or 240 hours later. Sections of brain and spinal cord were stained with the Carlsen-de Olmos cupric silver method. As reported previously, stained sections revealed degeneration in areas known to be innervated by small-diameter primary sensory fibers: the substantia gelatinosa of the spinal cord dorsal horn and spinal trigeminal nucleus, the solitary nucleus and tract, and the lateral borders of the area postrema. In addition, axon and terminal degeneration was observed in several discrete forebrain and hindbrain areas not previously associated with capsaicin-induced degeneration in either adult or neonatal rats: the inferior olive, the olivary pretectal nucleus, the interpeduncular nucleus, the suprachiasmatic nucleus, and the lateral septum/medial accumbens region. Furthermore, degenerating cell bodies were observed in the intrafascicular nucleus of the ventromedial midbrain tegmentum, in the supramammillary nucleus, and in the posterior hypothalamic area. Unilateral nodose ganglionectomy produced terminal staining on the denervated side very similar to that induced bilaterally by capsaicin. In addition, unilateral nodose ganglionectomy 1 month prior to capsaicin injection greatly attenuated staining in the ipsilateral nucleus of the solitary tract, confirming the hypothesis that capsaicin damages vagal sensory neurons innervating this nucleus. Capsaicin-induced damage in adult rats was long-lasting, since the second of two capsaicin treatments spaced 4.5 months apart produced no additional degeneration.     

Asymmetry of the olfactory system in the brain of the winter flounder,Pseudopleuronectes americanus

Adult flatfishes exhibit grossly asymmetric external morphology. Even the olfactory apparatus is asymmetric, being larger on the upward-facing side. We undertook the present study on the winter flounder, Pseudopleuronectes americanus, to examine whether the asymmetry of the peripheral olfactory system is maintained in its central organization. In winter flounder, the right olfactory organ, nerve, and bulb are larger than the contralateral counterparts. In addition, the right telencephalon is about 8% larger than the left. Horseradish peroxidase (HRP) and degeneration techniques were used to trace the central connections of the olfactory bulbs. Neurons afferent to the olfactory bulb occur bilaterally in the telencephalon and mesencephalic tegmentum. Afferent neurons are also present at the junction between the posterodorsal bulb and telencephalon, in the basal preoptic region, nucleus of the posterior tuber, locus coeruleus, raphe nucleus, and the contralateral bulb. Each olfactory bulb projects bilaterally to several restricted areas of the telencephalon, the posterodorsal neurons of the nucleus preopticus and the tuberal region, with ipsilateral connections being heavier in all areas. Corresponding to the differences in the peripheral olfactory apparatus, the central olfactory projections were also asymmetric. The right olfactory bulb projects to 2.6% of the ipsilateral telencephalon and 1.99% of the contralateral telencephalon. The left bulb projects to 1.8% of the ipsilateral and 0.6% of the contralateral telencephalic hemisphere. Thus the left telencephalon receives roughly equal olfactory input from the two sides, while the right telencephalon receives vastly more input from the right olfactory system. The asymmetry in the projections of the right and left bulbs may be due to differential postmetamorphic growth of the olfactory system on the two sides.     

An atlas of the primary visual projections in the brain of the chick Gallus gallus

The localisation of the primary visual centres in the chick mesencephalon and diencephalon was determined by autoradiographic anterograde transport and degeneration techniques. Strong visual projections were found in the tectum, lateral anterior thalamic nucleus, lateroventral geniculate nucleus, superficial synencephalic nucleus, external nucleus, ectomammillary nucleus, tectal grey, dorsolateral anterior thalamus, rostrolateral part, and the pretectal optic area. Weaker retinal projections were found in the ventrolateral thalamus, two subregions of the dorsolateral anterior thalamus, lateral part, diffuse pretectal nucleus, dorsolateral anterior thalamus, magnocellular part, and the hypothalamus. An atlas of the retinal projections was constructed from sections.     

Spatiotemporal Patterns of Pax3, Pax6, and Pax7 Expression in the Developing Brain of a Urodele Amphibian,Pleurodeles waltl

The onset and developmental dynamics of Pax3, Pax6, and Pax7 expressions were analyzed by immunohistochemical techniques in the central nervous system (CNS) of embryos, larvae, and recently metamorphosed juveniles of the urodele amphibian Pleurodeles waltl. During the embryonic period, the Pax proteins start being detectable in neuroepithelial domains. Subsequently, they become restricted to subsets of cells in distinct brain regions, maintaining different degrees of expression in late larvae and juvenile brains. Specifically, Pax6 is broadly expressed all along the urodele CNS (olfactory bulbs, pallium, basal ganglia, diencephalon, mesencephalic tegmentum, rhombencephalon, and spinal cord) and the developing olfactory organ and retina. Pax3 and Pax7 are excluded from the rostral forebrain and were usually observed in overlapping regions during embryonic development, whereas Pax3 expression is highly downregulated as development proceeds. Thus, Pax3 is restricted to the roof plate of prosomere 2, pretectum, optic tectum, rhombencephalon, and spinal cord. Comparatively, Pax7 was more conspicuous in all these regions. Pax7 cells were also found in the paraphysis, intermediate lobe of the hypophysis, and basal plate of prosomere 3. Our data show that the expression patterns of the three Pax genes studied are overall evolutionarily conserved, and therefore could unequivocally be used to identify subdivisions in the urodele brain similar to other vertebrates, which are not clearly discernable with classical techniques. In addition, the spatiotemporal sequences of expression provide indirect evidence of putative migratory routes across neuromeric limits and the alar–basal boundary. J. Comp. Neurol. 521:3913–3953, 2013. © 2013 Wiley Periodicals, Inc.     

Organization of the histaminergic system in the brain of the teleost, Trachurus trachurus

To accumulate phylogenetic information on the central histaminergic system, we investigated the histaminergic system in the brain of a teleost, the jack mackerel (Trachurus trachurus), using the indirect immunofluorescent method with antiserum against histamine. A small number of histamine-immunoreactive cell bodies were observed in the posterior hypothalamus around the posterior recess. Histamine-immunoreactive fibers innervated the telencephalon, diencephalon, tegmentum, and rostral part of the medulla oblongata. The immunoreactive fibers were very sparse or absent in the olfactory bulb, optic tectum, cerebellum, caudal part of the medulla oblongata, spinal cord, and hypophysis. Ascending fiber bundles were seen in the basal hypothalamus, supplying fiber collaterals to the telencephalon and diencephalon, whereas descending fibers were observed in the midline of the lower brainstem. These findings suggest that the central histaminergic system of the jack mackerel is homologous to those of mammals, reptiles, and amphibians, although poorly developed compared with them. The histamine-immunoreactive neuronal cell bodies found in the border area between the mesencephalon and rhombencephalon of the river lamprey were not detected in the brain of the jack mackerel.     

Starburst amacrine cells of the primate retina

A group of readily recognized amacrine cells were observed in Golgi-impregnated and flat-mounted macaque, baboon, and human retinas. These cells had roughly-circular or oval dendritic fields that were narrowly stratified within the inner plexiform layer (IPL). Most of these cells stratified in the inner half (sublamina b) of the IPL, and they had their somata in the ganglion-cell layer; a few stratified in the outer half (sublamina a) of the IPL and had their somata in the amacrine-cell layer. Typically, a single dendrite issued from the soma, and, after passing for 10 microns or so, gave rise to five or more radiate processes. As these processes neared the edge of the dendritic field they branched, turned, and became varicose. Most showed no evidence of an axon, although a few had a short process extending inward, toward the optic-fiber layer. Dendritic-field diameters were about 100 microns near the fovea and increased to about 350 microns in the peripheral retina. Mean somal diameter also increased slightly from near the fovea (7.8 microns) to the periphery (8.7 microns). Although the primate cells are smaller, and there are some minor differences in the form of the dendritic fields, these cells appear to be morphologically equivalent to the starburst amacrines of the rabbit retina, whose counterparts have also been observed in the retinas of rats and cats. Presuming that these cells correspond to the choline acetyltransferase immunoreactive primate cells described by Mariani and Hersh (J. Comp. Neurol. 267:269-280, '87), their overlap factor is about ten for the type whose somata lay in the ganglion-cell layer and about 0.25 for those whose somata lay in the amacrine-cell layer.     

Photothrombosis-induced ischemic neuronal degeneration in the rat retina

Here we describe a new model for inducing ischemic neuronal degeneration in the adult rat retina. Rose bengal dye was injected intravenously; then the retina was exposed to intense light which caused vascular photothrombosis resulting in acute degeneration of retinal neurons. By either light or electron microscopical criteria, the acute neurodegernative reaction was judged identical in pattern, cytopathological appearance, and time course to the excitotoxic type of reaction typically seen in the retina following exposure to exogenous glutamate. These results reinforce other accumulating evidence suggesting that ischemic CNS damage is mediated by glutamate or related excitotoxins. The advantages of this model of CNS ischemia include its noninvasiveness, ease of application, authentic simulation of vascular thrombosis, and accessibility for application of neuroprotective drugs.     

Auditory, electrosensory, and mechanosensory lateral line pathways through the forebrain in channel catfishes.

The forebrain auditory, electrosensory, and mechanosensory lateral line pathways in the channel catfish, Ictalurus punctatus, were examined by applying the fluorescent tracer DiI to 1) the auditory part of the torus semicircularis, 2) the electrosensory part of the torus semicircularis, 3) the lateral preglomerular nucleus, and 4) the anterior tuberal nucleus. Three distinct pathways ascend from the torus semicircularis to the telencephalon; they course through either 1) the lateral preglomerular nucleus of the posterior tuberculum, 2) the anterior tuberal nucleus of the hypothalamus, or 3) the central posterior nucleus of the dorsal thalamus. The anatomical data suggest that each of these ascending pathways carries information from more than one sensory modality. The lateral preglomerular nucleus receives an electrosensory input from nucleus electrosensorius in the diencephalon, but it also receives auditory and mechanosensory inputs directly from the torus semicircularis. The anterior tuberal and central posterior nuclei receive primarily auditory and mechanosensory, but also minor electrosensory, inputs. The efferent projections of the central posterior nucleus are presently unknown, but the lateral preglomerular and anterior tuberal nuclei project to nonoverlapping portions of the telencephalon. A cladistic analysis of these indirect torotelencephalic pathways reveals that 1) the pathway through the dorsal thalamus is probably a primitive character for gnathostomes, 2) a well-developed pathway through the posterior tuberculum is probably a derived character for actinopterygian fishes, 3) the pathway through nucleus electrosensorius is probably a derived character for catfishes and gymnotoid teleosts, and 4) auditory pathways through the hypothalamus probably evolved independently in catfishes and frogs.     

Parasol and midget ganglion cells of the human retina

Golgi-impregnated ganglion cells were studied in two flat-mounted human retinas. A number of different morphologic forms were observed, and those showing a thickly branching dendritic field with terminals that stratified within a narrow zone of the inner plexiform layer were selected for further investigation. When the dendritic field diameter of these cells was plotted against distance from the fovea, the scatter diagram showed two distinct clusters. At any given eccentricity, there was no overlap between the cell group with large dendritic fields and the group with small dendritic fields. Those with the larger dendritic fields also tended to have larger somas and thicker axons than the group with the smaller dendritic fields. The dendritic fields of both groups tended to be elongated, and the orientation and degree of this elongation were quantified by determining the best-fitting ellipse for each dendritic field. The degree of elongation was independent of eccentricity. The orientation of the dendritic field (major axis of the ellipse) of a cell did not appear to be independent of its position on the retina. To test whether the major axes tended to be directed toward any particular point on the retina, the positions of the cells on the retinal flat mount were transformed to relative positions on the retinal hemisphere, and the orientations of the dendritic fields were expressed in a spherical coordinate system for this hemisphere. A search was made for the position on the hemisphere which minimized the mean square deviation of the orientations from this point. The group with the large dendritic fields showed a significant tendency to be radially oriented toward a specific location on the retinal hemisphere, and that location lay within a few degrees of the fovea. Leventhal and Schall ('83) have reported a similar finding for ganglion cells of the cat retina. For the group with small dendritic fields, the retinal location that minimized the mean square deviation was also near the fovea; however, the set of orientations showed no greater tendency for mutual alignment than did a randomized set. The cell group with the large dendritic fields appears to correspond to Dogiel's (1891) type II cells, to Polyak's ('41) parasol cells, to the A cells of the monkey retina described by Leventhal et al. ('81), observed following HRP injection to the magnocellular layer of the LGN, and to the P alpha cells of the monkey retina, observed by Perry and Cowey ('81), following HRP uptake by cut axons of the optic nerve.(ABSTRACT TRUNCATED AT 400 WORDS)     

Hypothalamic and extrahypothalamic sauvagine-like immunoreactivity in the bullfrog (Rana catesbeiana) central nervous system

In the present study, immunocytochemistry and radioimmunoassay were used to investigate the presence of sauvagine in both hypothalamic and extrahypothalamic areas of the central nervous system (CNS) of the bullfrog (Rana catesbeiana) using a specific antiserum raised against synthetic non-conjugated sauvagine (SVG), a frog (Phyllomedusa sauvagei) skin peptide of the corticotropin-releasing factor (CRF) family. Sauvagine-immunoreactive (SVG-ir) bipolar neurons were found in the nucleus of the fasciculus longitudinalis medialis located in the rostral mesencephalic tegmentum. In the tectal mesencephalon, beaded SVG-ir fibres were present in the optic tectum, and in the torus semicircularis. Abundant SVG-ir varicose fibres were seen in the granulosa layer of the cerebellum, the nucleus isthmi, and the obex of the spinal cord. SVG-ir fibres were also seen by the alar plate of the rombencephalon. In the diencephalon, the antiserum stained parvocellular neurons of the preoptic nucleus (PON) which extended their dendrites into the cerebro-spinal fluid (CSF) of the third ventricle and projected their ependymofugal fibres to the zona externa (ZE) of the median eminence. Immunopositive fibres were also present in the medial forebrain bundle at the chiasmatic field, the posterior thalamus, the pretectal gray, and the ventrocaudal hypothalamus. In the telencephalon (forebrain), SVG-ir fibres were seen in the medial septum, the lateral septum, and the amygdala. The SVG immunoreactivity could not be detected after using the SVG antiserum previously immunoabsorbed with synthetic SVG (0.1 μM), but immunoblock of the antiserum with sucker (Catostomus commersoni) urotensin I (sUI), sole (Hippoglossoides elassodon) urotensin I, sucker CRF, rat/human CRF, or ovine CRF (0.1–10 μM) did not eliminate visualization of the immunoreactivity. In radioimmunoassay, the SVG antiserum did not crossreact with sUI, or the SVG fragments SVG_1–16, SVG_16–27, and SVG_26–34, but it recognized the C-terminal fragment SVG_35–40. Crossreaction with mammalian ovine CRF and rat/human CRF was negligible. Both hypothalamic and mesencephalic extracts gave parallel displacement curves to SVG. The results suggest the presence in the bullfrog brain of a SVG-like neuropeptide, i.e., a peptide of the CRF family, that either is SVG or shares high homology with the C-terminus of that peptide. The function of this neuropeptide in amphibians is not known at this time, but based on its anatomical distribution to the ZE it could affect the release of adrenocorticotropin (ACTH) or other substances from the amphibian pars distalis. Involvement of the SVG-like peptide in behavioural (forebrain), visual (thalamus-tegmentum mesencephali-pretectal gray-optic tectum), motor coordination (cerebellum), and autonomic (spinal) functions, as well as an undefined interaction with the CSF in the bullfrog seems likely. © 1996 Wiley-Liss, Inc.     

Connections of the lobus inferior hypothalami of the clearnose skate Raja eglanteria (Chondrichthyes)

The afferent and efferent connections of the lobus inferior hypothalami of the clearnose skate were demonstrated by the anterograde and retrograde transport of horseradish peroxidase. The main source of afferents is from the midbrain tegmentum and telencephalon. The major midbrain input is from cells of the ipsilateral nucleus tegmentalis lateralis and the caudal tegmental area. Another prominent, mostly ipsilateral, projection arises from nucleus F of the isthmic region. A few labeled cells also occur in the nucleus interpeduncularis, nucleus raphes superior, and lateral reticular formation. Afferents from the telencephalon arise from cells of the area preoptica, area superficialis basalis, striatum, nucleus septalis lateralis, and area subpallialis 1. Of the pallial structures, the pallium mediale and anterior as well as posterior subdivisions of the pallium dorsale pars centralis appear to have strong projections to the inferior lobe. Efferent connections of the inferior lobe consist of ascending and descending pathways. Fibers of the main ascending efferent pathway course within the basal forebrain bundle and distribute to subpallial areas. The descending efferent pathways course within the tractus lobobulbaris and tractus lobocerebellaris. Of these, the former is traceable to the level of the facial motor nucleus, issuing fibers enroute to the midbrain tegmentum and to the lateral reticular formation. The lobocerebellar tract courses dorsolateral to the lobobulbar tract, and its fibers terminate within the ipsilateral granular ridge of the rostral pole of the cerebellar corpus. There appears to be a topological organization of the inferior lobe connections. In general, pallial areas project mainly to the lateral subdivision of the inferior lobe nucleus at midlobic levels, whereas connections with the brainstem arise from or terminate within the dorsal and intermediate subdivisions at midlobic as well as caudal levels. The widespread ascending and descending connections indicate that the hypothalamic inferior lobe of the clearnose skate is a major relay center between the telencephalon and brainstem.     

Distribution of the limbic system-associated membrane protein (LAMP) in pigeon forebrain and midbrain

The limbic system-associated membrane protein (LAMP) is an adhesion molecule involved in specifying regional identity during development, and it is enriched in the neuropil of limbic brain regions in mammals but also found in some somatic structures. Although originally identified in rat, LAMP is present in diverse species, including avians. In this study, we used immunolabeling with a monoclonal antibody against rat LAMP to examine the distribution of LAMP in pigeon forebrain and midbrain. LAMP immunolabeling was prominent in many telencephalic regions previously noted as limbic in birds. These regions include the hippocampal complex, the medial nidopallium, and the ventromedial arcopallium. Subpallial targets of these pallial regions were also enriched in LAMP, such as the medial-most medial striatum. Whereas some telencephalic areas that have not been regarded as limbic were also LAMP-rich (e.g., the hyperpallium intercalatum and densocellulare of the Wulst, the mesopallium, and the intrapeduncular nucleus), most nonlimbic telencephalic areas were LAMP-poor (e.g., field L, the lateral nidopallium, and somatic basal ganglia). Similarly, in the diencephalon and midbrain, prominent LAMP labeling was observed in such limbic areas as the dorsomedial thalamus, the hypothalamus, the ventral tegmental area, and the central midbrain gray, as well as in a few nonlimbic areas such as nucleus rotundus, the shell of the nucleus pretectalis, the superficial tectum, and the parvocellular isthmic nucleus. Thus, as in mammals, LAMP in birds appears to be enriched in most known forebrain and midbrain limbic structures but is present as well in some somatic structures.     

Topographical distribution of acetylcholinesterase and butyrylcholinesterase in the diencephalon and mesencephalon of the garden lizard (Calotes versicolor)

The present report incorporates the histochemical mapping of acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) among the various nuclei and fiber tracts of the diencephalon and mesencephalon of Calotes veriscolor. The various nuclei, for both enzymes, present varying degrees of staining, ranging from negative nuclei, on the one hand, to mild and intense staining on the other hand. Almost all of the fiber tracts reveal intense activity in BChE preparations, while they demonstrate mild and moderate activities for AChE. The nature of the various nuclei in relation to enzymatic patterns is discussed.     

Ibotenate-induced neuronal degeneration in immature rat brain

Stereotaxic microinjections of the excitotoxin, ibotenic acid, were made into the striatum, hippocampus or cerebellum of the immature (7-day-old) rat. Two days later, pups were decapitated and the brains processed for light microscopic examination or neurochemical analyses. 10 micrograms ibotenate caused a complete loss of nerve cell bodies throughout the striatum and hippocampus while intracerebellar injections produced no detectable damage. In the striatum, catecholamine histofluorescence was abolished and dopamine uptake severely reduced, indicating also a loss of afferent nerve terminals. Co-injection of 10 micrograms ibotenate with equimolar amounts of the selective amino acid antagonist, (-)-2-amino-7-phosphonoheptanoic acid, resulted in the protection of both striatal cell bodies and dopaminergic nerve terminals. The neurotoxic properties of ibotenate described here are in marked contrast to those of kainic acid, a related excitotoxin. Differences in the ontogenetic pattern of receptors which mediate neurodegenerative events may account for the pharmacological and regional selectivity and the partial lack of axon-sparing properties of ibotenic acid lesions in the immature brain.     

Hypoxia induces differential changes of dopamine metabolism in mature and immature mesencephalic and diencephalic cell cultures

Summary. Perinatal hypoxia is known as a high risk factor for the development of long-lasting abnormalities in dopaminergic system. The early developmental alterations of dopamine (DA) metabolism induced by hypoxia could contribute to these abnormalities. To understand the hypoxia-induced changes of intra- and extracellular dopamine levels and its main metabolites, 3,4-dihydroxyphenylacetic acid (DOPAC) and homovanillic acid (HVA), in immature dopaminergic neurons, we compared these changes in rat mesencephalic and diencephalic cell cultures on day in vitro (DIV) 2 (immature cells), DIV 8 and DIV 13 (mature cells). Cell cultures were exposed to an oxygen-free gas mixture in a Billups chamber for 2–4 hours. Mature cell cultures responded to hypoxia with an increase of DA levels in the cells and in the medium during the first 45 min (by an average of 57 and 114% respectively). Thereafter, DA levels decreased, and returned to the baseline within the next 30 min. The cellular DA levels continued to decrease up to 15% of the baseline during 255 min hypoxia whereas the extracellular DA content stabilized at the prehypoxic levels. Immature cell cultures (DIV 2) in contrast to mature ones, were unable to maintain normal extracellular DA levels during hypoxia and showed a decrease of the cellular and extracellular levels to 50% of the prehypoxic levels. DOPAC and HVA changes mimick, however, at a lower level, the pattern of DA changes during the exposure to hypoxia. In principle, in the diencephalic cell culture similar effects of hypoxia exposure on the investigated parameters were found (studied during 0–120 min). The present study demonstrates that mature and immature dopaminergic cells differ in the regulation of the extra- and intracellular DA levels during hypoxia. In immature cells the low synthetic capacity of tyrosine hydroxylase and the deficient capacities of the transport and storage processes result in decreased extracellular DA levels. This could be an important factor for the long-term modulation of the expression of tyrosine hydroxylase and subsequent long-term behavioral and/or neurological abnormalities induced by perinatal hypoxia. Received June 8, 1998; accepted July 21, 1998     

Transneuronal retrograde degeneration in the cat retina following neonatal ablation of visual cortex

Bilateral removal of the cortical visual areas in newborn cats produced degeneration of retinal ganglion cells. Measurements of over 4,000 cells and calculation of neuron densities from sample areas of retina in the adult show that the medium sized cell population in peripheral retina is reduced by 68%, whereas the populations of small and large cells are not affected. The degeneration is greater in peripheral retina than in area centralis.