Development of histamine-immunoreactive neurons in the rat brain

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

Comparative neuroanatomy of the histaminergic system in the brain of the frog Xenopus laevis

The distribution of the histaminergic neuronal system in the brain of the clawed frog Xenopus laevis was mapped with an antiserum against carbodiimide-fixed histamine and compared to that in mammals. The histamine-immunoreactive cell bodies were located in a small area of the posterolateral hypothalamus, close to the dorsal infundibular nucleus, which contains catecholaminergic and serotonergic neurons. This area may be homologous to the tuberomammillary nucleus in mammals. A thick process extended from each cell between the ependymal cell layer and terminated in the ventricle lumen. The number of histaminergic cell bodies in adult Xenopus brain was relatively low, as compared with the mammalian brain. Preliminary analysis of adjacent sections stained with antisera against GABA or serotonin indicated that the histamine cells were not immunoreactive for these. The pathways and distribution of histaminergic fibers in Xenopus brain showed many similarities to mammals. The densest fiber networks were present in the medial basal forebrain, particularly in the medial amygdala and septum. A distinct cluster of fibers was concentrated around the cell bodies of nucleus accumbens. In most pallial areas, the density was moderate to low. In the primordial piriform cortex and the striatum, very few fibers were seen. In diencephalon, highest fiber densities were found in the anterior and ventral thalamus and posterior and lateral hypothalamus. In hindbrain, the density was highest in the medullary central gray, as in some mammals. The results suggest that the general pattern of the histaminergic system in vertebrate brain is conserved from amphibians to mammals.     

Histamine-immunoreactive neurons and their innervation of visual regions in the cortex,tectum, and thalamus in the primate Macaca mulatta

The histaminergic system is involved in the control of arousal in the brain and may impact significantly on visual processing. However, little is known about the histaminergic innervation of visual areas, or the histamine system in the primate brain, in general. We examined in Macaca mulatta the location of histamine-immunoreactive neurons and the innervation of important cortical and subcortical visual areas by histamine-immunoreactive axons. Brain sections were treated with an antibody to histamine and processed with standard immunohistological procedures. Histamine-immunoreactive neurons (20–45 μm in diameter) were localized bilaterally in the hypothalamus, particularly in ventral, lateral, posterior, and perimammillary hypothalamic areas. These hypothalamic cells appear to provide the sole neural source of histamine in the macaque brain. A plexus of varicose histamine-immunoreactive axons was present throughout the superior colliculus, the dorsal and ventral lateral geniculate nuclei of the thalamus, the reticular nucleus of the thalamus, the lateral posterior/pulvinar complex, and the visual cortex, including areas 17, 18, and the nearby extrastriate cortex. The axons nearly homogeneously innervated every region and layer in these structures, except for an increase in density in layer 1 of the visual cortex and in the superficial-most layers of the superior colliculus. Histaminergic axons broadly innervated every visual region examined. In comparison with the other aminergic and the cholinergic projection systems, which show considerable projection specificity, the histaminergic projection exhibited great homogeneity. The breadth of the distribution of histaminergic axons ensures that virtually all levels of visual processing in the primate can be influenced, either directly or indirectly, by the neuromodulatory effects of histamine. © 1996 Wiley-Liss, Inc.     

Subicular efferents to histaminergic neurons in the posterior hypothalamic region of the rat studied with PHA-L tracing combined with histidine decarboxylase immunocytochemistry.

The projections from the subiculum to histaminergic cells in the posterior hypothalamic region of the rat were studied by means of anterograde neuroanatomical tracing with Phaseolus vulgaris-leucoagglutinin (PHA-L) combined with histidine decarboxylase (HDC)-immunohistochemistry. PHA-L was injected at various loci along the dorsoventral and proximodistal axes of the subiculum. This resulted in labeling of the fornix and of terminal plexuses at various locations in the diencephalon and the mammillary body. Following deposition of PHA-L in the proximal part of the dorsal subiculum, labeled fibers in the posterior hypothalamus were confined to the mammillary nuclei, whereas after injections of PHA-L in the distal part of the dorsal subiculum and the entire ventral subiculum, labeled fibers were also present in clusters of histaminergic cells located around the mammillary nuclei. The density of the PHA-L labeled fibers within these clusters increased from low to moderate in association with a shift of the injection sites from dorsal to ventral and from proximal to distal parts of the subiculum, i.e., the highest fiber labeling was seen after injections of PHA-L in the distal part of the ventral subiculum. In the latter experiments, PHA-L labeled fibers reached HDC-immunoreactive neurons in the tuberal magnocellular nucleus, the deepest layer of the caudal magnocellular nucleus, the two bridges of histaminergic cells in the posterior hypothalamus, and the histaminergic neurons scattered in the supramammillary region. A few labeled fibers invaded the postmammillary caudal magnocellular nucleus. The presence of varicosities on the PHA-L labeled fibers in close proximity to the cell bodies and dendrites of the histaminergic neurons suggest the existence of synaptic contacts.     

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.     

Histamine-immunoreactive nerve fibers in the rat pineal gland: evidence for a histaminergic central innervation

An immunohistochemical method that utilizes carbodiimide as a fixative and antisera directed against histamine was applied to investigate the location of histamine in the rat pineal complex. Numerous histamine-immunoreactive cell bodies were observed in different subdivisions of the tuberomammillary nucleus of the posterior hypothalamus, and a few cell bodies were present in the posterior and dorsal part of the periventricular hypothalamic nucleus. Histamine-immunoreactive fibers were observed to leave the posterior hypothalamus in various directions of which one dorsally projecting tract was followed in the periventricular area of the caudal diencephalon to the epithalamus. Several histamine-immunoreactive nerve fibers of this tract continued through the posterior commissure directly into the deep pineal gland. A few immunoreactive fibers were also observed in the habenular commissure. In midsagittal sections, histamine-immunoreactive nerve fibers were observed to enter the pineal stalk from the deep pineal gland. Most of histamine-immunoreactive fibers in the stalk continued towards the superficial pineal gland, but their number decreased in more distal locations of the stalk, indicating that some fibers terminate in the stalk as well. A few fibers were found to terminate in the most rostral part of the superficial pineal gland. The immunoreactive nerve fibers in the epithalamus and pineal complex were endowed with prominent varicosities. Taken together, these results indicate that histaminergic nerve fibers, originating from the posterior hypothalamus, project to the pineal complex of the rat. Histamine must therefore be considered a putative neurotransmitter contained in the central innervation of the pineal gland, but its function in pineal physiology has so far not been elucidated.     

Histaminergic system in the tree shrew brain

This study mapped the histamine-immunoreactive neuronal system in the brain of the tree shrew (Tupaia belangeri) and compared its structure with that of the rat and guinea pig. The histamine-containing cell bodies lay in the posterior ventral hypothalamus in the tuberomammillary complex, as in the rodents. The morphology of this complex resembled that of the rat. The histaminergic axons projected to nearly all parts of the brain. The main ascending bundle ran ventromedially: the densest innervation was found in the ventral hypothalamus, preoptic area, septum, medial part of nucleus accumbens, and bed nucleus of the stria terminalis. High fiber densities were present in the amygdaloid nuclei and claustrum. Another pathway ran dorsomedially along the periventricular hypothalamus and sent fibers to all parts of the diencephalon. Part of these fibers followed the central gray to the midbrain and spread laterally below the inferior colliculus. Another descending pathway ran through the interfascicular and medial raphe nuclei to meet the pontine central gray. The densest fiber networks were seen in the dorsal tegmental and parabrachial nuclei, and around the locus coeruleus. Also the substantia nigra, interpeduncular and mesencephalic reticular nuclei, colliculi, and vestibular and raphe nuclei received a dense histaminergic innervation. The organization of the fibers in the tree shrew brain resembled more that in the guinea pig than that in the rat. As compared with the guinea pig, more fibers were present, particularly in the globus pallidus, central thalamus, and deep cerebellar nuclei. No fibers were seen in the outer layer of the piriform cortex. In Tupaia, a laminar organization of the fibers was evident in the hippocampus, in contrast to the rodents. Also, a dense periventricular fiber plexus was prominent.     

Histamine-immunoreactive neurons of the tuberomammillary nucleus are innervated by alpha-melanocyte stimulating hormone-containing axons. Generation of a new histamine antiserum for ultrastructural studies

Leptin regulates the release of histamine in the hypothalamus, however, the histaminergic neurons contain few leptin receptors. To reveal that alpha-melanocyte stimulating hormone (alpha-MSH)-containing neurons of the arcuate nucleus may mediate the effects of leptin to the histaminergic neurons, we studied the putative innervation of histamine-immunoreactive (IR) neurons by alpha-MSH-containing axons using double-labeling immunocytochemistry. In order to analyze the relationship of alpha-MSH- and histamine-IR elements, we also generated an antiserum against histamine that is compatible with acrolein-based fixatives commonly used for immuno-electron microscopic studies. The apposition of alpha-MSH-IR axons to histaminergic neurons was observed in all five subnuclei of the tuberomammillary nucleus. Both axo-somatic and axo-dendritic contacts were found. At the ultrastructural level, silver-intensified colloidal gold particles identified the histaminergic neurons, whose ultrastructure was well preserved after fixation with acrolein demonstrating that the new antiserum is a useful tool for the ultrastructural examination of the histaminergic system. The histamine-IR cells received synaptic inputs from alpha-MSH-IR axon terminals visualized by diaminobenzidine. These data indicate that alpha-MSH-synthesizing neurons innervate histaminergic neurons in the tuberomammillary nucleus and may relay the hormonal influence of leptin to the histaminergic system.     

Histaminergic neurons in the sheep diencephalon

The distribution of histaminergic neurons in the sheep brain was studied by immunohistochemistry by using antibodies raised against histamine. For the first time in this species, the presence of histamine-immunoreactive neurons was described in the caudal diencephalon, around the mammillary bodies, and in the tuberomammillary area. The general pattern of distribution of these neurons was similar to that described previously in other species, i.e., rodents and humans. The distribution in the five neuronal groups described in rodents was not easy to demonstrate in sheep, because the boundaries between each group were not clear. The labeled neurons appeared to form a continuous cell system, as in humans. Numerous histamine-immunoreactive mast cells were found in the habenula and the thalamus. Histamine-immunoreactive fibers were found in almost all of the structures studied. The highest density of fibers was seen in the tuberomammillary area, from which dense bundles of fibers ran rostrally and dorsally along the third ventricle in a parasagittal plane. Numerous immunostained fibers were found close to the wall of the ventricles; some of them appeared to reach the cerebrospinal fluid through the ependymal cell layer. Some fibers were also observed in the optic tract, and the lowest density was found in the supraoptic and paraventricular nuclei. These results should be useful for developing further physiological studies on the role of histaminergic neuronal systems in sheep. J. Comp. Neurol. 400:317–333, 1998. © 1998 Wiley-Liss, Inc.     

An altered histaminergic innervation of the substantia nigra in Parkinson's disease

The central histaminergic system is one of the subcortical aminergic projection systems involved in several regulatory functions. The central dopaminergic and histaminergic systems interact extensively, but little is known about the histaminergic system in diseases affecting the dopaminergic neurons. The distribution of histaminergic fibers in the substantia nigra (SN) in postmortem brain samples from patients suffering from Parkinson's disease (PD) and normal controls was examined with a specific immunohistochemical method. Direct connections between dopaminergic neurones and histaminergic fibers were observed. Histamine in human SN was stored in fibers and varicosities. Sites of histamine formation were examined by l-histidine decarboxylase in situ hybridization. In both normal and PD brains HDC mRNA was found only in posterior hypothalamus and not in SN. The presence of histaminergic innervation of the human substantia nigra pars compacta (SNc) and reticulata (SNr), paranigral nucleus, radix of oculomotor nerve, and parabrachial pigmented nucleus was demonstrated. The density of histaminergic fibers in the middle portion of SNc and SNr was increased in brains with PD. In PD the morphology of histaminergic fibers was also altered; they were thinner than in controls and had enlarged varicosities. An increase of histaminergic innervation may reflect a compensatory event due to deficiency of, e.g., dopamine or a putative fiber growth inhibitory factor. Whether the changes seen in histaminergic fibers in PD are primary or secondary remains to be investigated.     

Immunohistochemical localization of avian pancreatic polypeptide-like immunoreactivity in the rat hypothalamus

The distribution of avian pancreatic polypeptide-like (APP) immunoreactivity within the rat hypothalamus was investigated with the indirect immunoperoxidase method. APP immunoreactive perikarya are found in largest numbers in the retrochiasmatic area, the arcuate nucleus, and the supracommissural portion of the interstitial nucleus of the stria terminalis. Small clusters of immunoreactive neurons are also consistently observed in the ventral aspect of the medial preoptic area and lateral hypothalamic area, immediately dorsolateral to the optic chiasm and tracts. These neurons are apparent in all animals but are more intensely strained and occur in larger numbers following colchicine pretreatment. Other immunoreactive neurons are visible only in colchine-treated rats and are scattered throughout the anterior and lateral hypothalamic areas and the supramammillary nucleus. Immunoreactive axons and terminal fields present an extensive and highly characteristic distribution throughout the hypothalamus, which in many instances exhibits differential distribution within specific subfields of hypothalamic nuclei and areas. The heaviest concentrations of APP immunoreactive axons are present in the periventricular nucleus throughout the rostrocaudal extent of the hypothalamus, the ventrolateral portion of the suprachiasmatic nucleus, the retrochiasmatic area, the parvocellular paraventricular nucleus, the ventral supraoptic nucleus, the perifornical nucleus, the ventral dorsomedial nucleus, and the arcuate nucleus. Moderate plexuses of immunoreactive fibers are also present in the medial preoptic area, the anterior and lateral hypothalamic areas, the nucleus circularis, the median eminence, and the ventral premammillary area. Other areas, such as the ventromedial nucleus, contain virtually no immunoreactive axons but are encapsulated by a dense plexus of immunoreactive terminals. The distribution of a major component of APP immunoreactive fibers exhibits a marked similarity to that of previously described norepinephrine-containing hypothalamic afferents. Other groups of APP immunoreactive perikarya and fibers appear to represent components of intrinsic diencephalic systems.     

Histochemical mapping of dopamine neurons and fiber pathways in dog mesencephalon

A topographic mapping of dopamine (DA)-containing neurons and fibers was done mainly in the mesencephalon of the dog using the fluorescent histochemical technique of Falck and Hillarp. The extensive DA neuron system was found to be located in the ventral and medial regions of the mesencephalon; the pars compacta of the substantia nigra, the area almost corresponding to the ventral tegmental area of Tsai (hich consists of three groups, a caudal, the nucleus parabrachialis pigmentosus, a ventral, the nucleus paranigralis and a rostral, the caudal part of the nucleus tegmentalis gventralis of Tsai), the nucleus linearis of the raphe, and the mesencephalic reticular formation. The nigro-neostriatal projection can be traced in the non-treated or nialamide plus L-dopa treated puppies without the lesion-degeneration technique. Most fibers arising from these DA cell groups assemble at the prerubral area and ascend just dorsal to the medial forebrain bundle. Most fibers turn laterally at the lateral hypothalamus and enter the neostriatum via the dorsal part of the subthalamic nucleus, the zona incerta and the capsula interna. These findings show that the distribution of DA neurons and the nigro-neostriatal pathway are fundamentally similar to those in other mammals. In this study, the processes of the nigral and paranigral DA neurons have been demonstrated to project into the pars reticulata in the dog.     

Organization of the histaminergic system in the brain of the turtle Chinemys reevesii

To accumulate phylogenetic information on the central histaminergic system, we investigated the histaminergic system in the brain of the Reeves turtle, Chinemys reevesii, using the indirect immunofluorescent method with antiserum against histamine. Histaminergic neuronal cell bodies were found exclusively in the posterior part of the ventral hypothalamus. Histaminergic varicose fibers innervated almost all parts of the turtle brain, but tended to be concentrated in several areas. Very dense innervation was observed in the medial part of the telencephalon, ventrolateral part of the hypothalamus, nucleus habenularis lateralis, and ventromedial part of the tegmentum. Medium density of innervation was seen in the olfactory bulb, nucleus medialis amygdalae, and tectum. Only a few fibers were detected in the lateral part of the telencephalon, dorsal part of the hypothalamus, thalamus, rhombencephalon, and spinal cord. The main ascending fibers were observed in the lateral part of the hypothalamus, sending dense fiber bundles to the cortices dorsomedialis and medialis and nucleus habenularis lateralis. Descending fibers appeared to run in the ventral tegmental area, passing through the dorsal and ventral parts of the midline of the brain stem to the spinal cord. These findings indicate that the general morphological features of the histaminergic system in the turtle brain are similar to those in the mammalian and frog brains.     

Neurotensin in the ventromedial mesencephalon of the rat: Anatomical and functional considerations

Neurotensin is an endogenous neuropeptide that fulfills some of the criteria for a neurotransmitter in the mammalian central nervous system. It exists in high concentrations in the ventral tegmental area and adjacent midline nuclei of the ventromedial mesencephalon, and recent microinjection studies have demonstrated that neurotensin can act in this brain region to produce both a decrease in colonic temperature, and an increase in spontaneous motor activity. In this study it was found that hypothermia was most successfully evoked following neurotensin injection along the midline of the ventral mesencephalon, corresponding to the nucleus linearis centralis. In contrast, behavioral hyperactivity was produced with greatest consistency in the ventral tegmental area, corresponding to the nucleus paranigralis and nucleus parabrachialis pigmentosus. However, in its caudal aspect, the nucleus paranigralis was found unresponsive to neurotensin. Behavioral hyperactivity was also observed after neurotensin injection along the midline into the nucleus interfascicularis. Only injections made into the nucleus linearis rostralis produced hypothermia and hyperactivity in the same rat. This distribution of neurotensin-responsive nuclei corresponded to the distribution of neurotensin containing perikarya and fibers. With the exception of the nucleus interfascicularis, neurotensin-containing neurons were distributed throughout the rostral portion of the ventromedial mesencephalon, the nucleus parabraehialis pigmentosus containing the greatest density. However, in the caudal portion, neurotensin neurons were found almost exclusively in the nucleus linearis centralis. Neurotensin-containing fibers were of greatest density in the nucleus interfascicularis and the nucleus linearis centralis. Considering the known capacity of neurotensin to activate dopamine neurons in the ventromedial mesencephalon, and the partial mediolateral topographical distribution of dopaminergic projections from this region to the limbic forebrain, it is possible that neurotensin may be activating two distinct populations of dopamine neurons to produce hypothermia and behavioral hyperactivity.     

Altered distribution of dopaminergic neurons in the brain of L1 null mice

Dopaminergic neurons of the mouse mesencephalon originate in the ventricular zone and migrate radially along radial glia then tangentially along nerve fibers that express the neural cell adhesion molecule L1 to form the substantia nigra (A9 group) and ventral tegmental area (VTA) (A10 group). The role of L1 in migration of dopaminergic neuronal precursors was investigated in L1 knockout mice by tyrosine hydroxylase (TH) immunostaining. An altered rostrocaudal distribution of dopaminergic neurons was observed within the substantia nigra and VTA of L1-minus mice. In L1-minus mice at postnatal day 0, TH-positive cells were present abnormally in the dorsomedial mesencephalon, suggesting impaired migration. Axons projecting from the substantia nigra to the caudate putamen also exhibited an abnormal targeting pattern. There was no evidence of dopaminergic cell loss in the mutant SN. Abnormal localization of dopaminergic neurons in L1-minus mice was also evident in the zona incerta of the thalamus (A13 group), and the arcuate (A12) and periventricular nucleus (A14) of the hypothalamus. Cell bodies and axons in the substantia nigra, VTA, and hypothalamus of wild type mouse embryos expressed L1. These results suggested that L1 plays an important developmental role in the organization of dopaminergic neuronal cell groups in the mesencephalon and diencephalon.     

Neurotensin-like Peptides in the CNS of Lampreys: Chromatographic Characterization and Immunohistochemical Localization with Reference to Aminergic Markers

Neurotensin (NT)-like peptides in the CNS of the lamprey Lampetra fluviatilis were studied by radioimmunoassay (C-terminal specific NT antiserum), reverse-phase HPLC and immunohistochemistry. Multiple peaks of NT-immunoreactive (-ir) material were observed upon HPLC, of which a major peak eluted in the position of bovine NT. Immunofluorescence histochemistry showed that a monoclonal antibody recognizing the N-terminal (1 - 11) fragment of NT, as well as two polyclonal NT antisera labelled a large number of cell bodies in the periventricular area of hypothalamus, including the postinfundibular commissural nucleus and the ventral and dorsal hypothalamic nuclei. Additional groups of NT-ir cells were observed in the preoptic nucleus, the postoptic commissural nucleus, the mesencephalic tegmentum (L.fluviatilis), and in the spinal cord (L.fluviatilis and Ichtyomyzon unicuspis). Dense NT-ir fibre plexuses were present in the caudal hypothalamus, corpus striatum, ventral mesencephalon, and in the dorsal horn and lateral margin of the spinal cord. At the ultrastructural level the lateral spinal margin showed NT-ir terminal structures, which in most cases were not associated with synaptic specializations, although occasional synaptic contacts with unlabelled elements were found. The relation between NT-ir and monoamine-containing cells was examined with immunofluorescence double-staining, using antisera to tyrosine hydroxylase (TH), 5-hydroxytryptamine (5-HT), and histamine respectively. In the periventricular nuclei of hypothalamus numerous TH-, 5-HT-, as well as histamine-ir cells were located in close association with NT-ir cells, but none of the aminergic markers could be detected within NT-ir neurons. The chemical properties as well as the anatomical distribution of lamprey NT-like peptides show several similarities with those present in mammals, suggesting that NT-containing neuronal systems in the CNS developed early in vertebrate phylogeny.     

Comparative distribution of neuropeptide-immunoreactive systems in the brain of the green molly, Poecilia latipinna

The comparative distribution of peptidergic neural systems in the brain of the euryhaline, viviparous teleost Poecilia latipinna (green molly) was examined by immunohistochemistry. Topographically distinct, but often overlapping, systems of neurons and fibres displaying immunoreactivity (ir) related to a range of neuropeptides were found in most brain areas. Neurosecretory and hypophysiotrophic hormones were localized to specific groups of neurons mostly within the preoptic and tuberal hypothalamus, giving fibre projections to the neurohypophysis, ventral telencephalon, thalamus, and brain stem. Separate vasotocin (AVT)-ir and isotocin (IST)-ir cells were located in the nucleus preopticus (nPO), but many AVT-ir nPO neurons also displayed growth hormone-releasing factor (GRF)-like-ir, and in some animals corticotrophin-releasing factor (CRF)-like-ir. The main group of CRF-ir neurons was located in the nucleus recessus anterioris, where coexistence with galanin (GAL) was observed in some cells. Enkephalin (ENK)-like-ir was occasionally present in a few IST-ir cells of the nPO and was also found in small neurons in the posterior tuberal hypothalamus and in a cluster of large cells in the dorsal midbrain tegmentum. Thyrotrophin-releasing hormone (TRH)-ir cells were found near the rostromedial tip of the nucleus recessus lateralis. Gonadotrophin-releasing hormone (GnRH)-ir cells were present in the nucleus olfactoretinalis, ventral telencephalon, preoptic area, and dorsal midbrain tegmentum. Molluscan cardioexcitatory peptide (FMRF-amide)-ir was colocalized with GnRH-ir in the ganglion cells and central projections of the nervus terminalis. Melanin-concentrating hormone (MCH)-ir neurons were restricted to the tuberal hypothalamus, mostly within the nucleus lateralis tuberis pars lateralis, and somatostatin (SRIF)-ir neurons were numerous throughout the periventricular areas of the diencephalon. A further group of SRIF-ir neurons extending from the ventral telencephalon into the dorsal telencephalon pars centralis also contained neuropeptide Y (NPY)-, peptide YY (PYY)-, and NPY flanking peptide (PSW)-like-ir. These immunoreactivities were, however, also observed in non-SRIF-ir cells and fibres, particularly in the mesencephalon. Calcitonin gene-related peptide (CGRP)-like-ir had a characteristic distribution in cells grouped in the isthmal region and fibre tracts running forward into the hypothalamus, most strikingly into the inferior lobes. Antisera to cholecystokinin (CCK) and neurokinin A (NK) or substance P (SP) stained very extensive, separate systems throughout the brain, with cells most consistently seen in the ventral telencephalon and periventricular hypothalamus. Broadly similar, but much more restricted, distributions of cells and fibres were seen with antisera to neurotensin (NT) and vasoactive intestinal peptide (VIP).(ABSTRACT TRUNCATED AT 400 WORDS)     

Efferent projections of the retrorubral nucleus to the substantia nigra and ventral tegmental area in cats as shown by anterograde tracing

The aim of the present study was to determine whether the retrorubral nucleus projects to the dopaminergic nuclei in the ventral midbrain of the cat. For this purpose, injections of biotinylated dextran-amine or Phaseolus vulgaris-leucoagglutinin were placed into the retrorubral nucleus under stereotaxic guidance. The tracers were visualized by means of (immuno) histochemical procedures. In addition, tyrosine hydroxylase immunohistochemistry was used to evaluate the location of the injection sites and the distribution of the anterogradely labeled fibers. Both tracers reveal the same topography of labeled fibers in the ventral mesencephalon. Labeled fibers with varicosities were found ipsilaterally in the substantia nigra pars compacta, the substantia nigra pars lateralis, the ventral tegmental area and, contralaterally, in the substantia nigra pars compacta, the ventral tegmental area, and the retrorubral nucleus. A considerable number of labeled axons with varicosities were observed to be wrapped around the dendrites and perikarya of tyrosine hydroxylase-positive neurons in these areas. The present results are discussed in view of the possible role of the A8 dopaminergic cell group in the coordination of A9 nigrostriatal and A10 mesolimbic systems, as well as in the progressive pathology seen in patients suffering from Parkinson's disease.     

Neuropeptide tyrosine in the brain of the African lungfish,Protopterus annectens: Immunohistochemical localization and biochemical characterization

Lungfishes, which share similarities with both fishes and amphibians, represent an interesting group in which to investigate the evolutionary transition from fishes to tetrapods. In the present study, we have investigated the localization and biochemical characteristics of neuropeptide Y (NPY)-immunoreactive material in the central nervous system of the African lungfish, Protopterus annectens. NPY-immunoreactive cell bodies were found in various regions of the brain, most notably in the telencephalon (septal area, ventral striatum, and nucleus accumbens), in the diencephalon (preoptic nucleus, periventricular region of the hypothalamus, and ventral thalamus), and in the tegmentum of the mesencephalon. A strong immunoreaction was also detected in cell bodies of the nervus terminalis. Immunoreactive nerve fibers were particularly abundant in the ventral striatum, the nucleus accumbens, the diagonal band of Broca, the hypothalamus, and the mesencephalic tegmentum. Positive fibers were also seen in the median eminence and in the neural lobe of the pituitary. The NPY-immunoreactive material localized in the brain and pituitary was characterized by combining high-performance liquid chromatography (HPLC) analysis and radioimmunological quantitation. The displacement curves obtained with synthetic porcine and frog NPY and serial dilutions of brain and pituitary extracts were parallel. Reversed-phase HPLC analysis of telencephalon, diencephalon, and pituitary extracts resolved a major NPY-immunoreactive peak that coeluted with frog NPY. The similarity between the distribution of NPY-containing neurons and the biochemical characteristics of the immunoreactive peptide in the brain of lungfish and frog strongly favors a close phylogenetic relationship between dipnoans and amphibians. © 1995 Wiley-Liss, Inc.     

Immunocytochemical analysis of the dopamine system in the forebrain and midbrain of Raja radiata: evidence for a substantia nigra and ventral tegmental area in cartilaginous fish

The distribution of dopamine-containing cell somata and fibers in the forebrain and midbrain of a cartilaginous fish, Raja radiata, was investigated by means of antibodies directed against dopamine. Many small dopamine immunoreactive neurons are distributed throughout the telencephalon, including the olfactory bulbs. Within the diencephalon and particularly in the hypothalamus, i.e., in the nucleus preopticus, nucleus suprachiasmaticus, the paraventricular organ, lateral hypothalamic area, recessus mamillaris, and nucleus tuberculi posterioris, numerous cell somata stain for dopamine. In the mesencephalon, two distinct cell masses are found, which on the basis of their immunoreactivity for dopamine and their location, may be homologous to the substantia nigra and ventral tegmental area of other vertebrates. Dopamine immunoreactive fibers are found in the glomeruli of the olfactory bulbs, in ventral portions of the telencephalon, where a dense dopaminergic plexus innervates the area superficialis basalis and striatum, and in the diencephalon, where the inferior lobe is the most densely innervated structure. In the mesencephalon, the dopamine immunoreactive fibers are confined predominantly to the periventricular zone and lateral portions of the tectum. We conclude that much of the dopaminergic system in Raja radiata is strikingly similar to that seen in amniotes.