Tyrosine hydroxylase-immunoreactive cell groups in the brain of the teleost fishGnathonemus petersii

Different antibodies against tyrosine hydroxylase (TH) were used to obtain detailed information about the distribution, morphology and chemical differentiation of catecholaminergic neurons in the highly differentiated brain of the electric mormyrid fishGnathonemus petersii. The results show that the distribution of catecholaminergic neurons is much more widespread than was previously thought on the basis of dopamine and noradrenaline immunohistochemistry. Tyrosine hydroxylase-immunoreactive neurons were observed not only in clearly dopaminergic regions (the suprachiasmatic nucleus, the magnocellular hypothalamic nucleus and the area postrema) and noradrenergic cell groups (the locus coeruleus and inferior reticular cell group), but also in regions that do not, or only fragmentarily, display dopamine or noradrenaline immunoreactivity, including the ventral and intermediate telencephalon, the anterior and posterior preoptic cell group, the ventromedial thalamus, the pretectal region and the nucleus of the solitary tract, suggesting that they either represent depleted dopaminergic cell groups orl-dihydroxy phenlalanine- producing nuclei. Most TH-immunoreactive neurons are rather small (<10 μm) and have only a few slender processes, but neurons in the magnocellular hypothalamic nucleus and the inferior reticular formation are multipolar and larger (10–20 μm), while those of the locus coeruleus are even more than 20 μm in diameter. The hypothalamic paraventricular organ, which is strongly dopamine and noradrenaline immunoreactive, displays minimal TH immunoreactivity, suggesting that its cerebrospinal fluid-contacting neurons do not synthesize catecholamines, but acquire them from external sources.

Comparison with other teleosts shows that the catecholaminergic system in the brain of Gnathonemus is similarly organized as inCarassius, Gasterosteus, Anguilla andAperonotus, with some variations that may partly be due to technical reasons, and partly reflect true species differences. However, TH-immunoreactive neurons in the midbrain tegmentum were not observed, confirming previous conclusions that a major difference between teleosts and mammals concerns the absence of dopaminergic midbrain groups and correlated mesencephalo-telencephalic projections in teleosts.     

Distribution and morphology of the catecholaminergic neural elements in the human hypothalamus

Previous studies have demonstrated that catecholaminergic, tyrosine hydroxylase (TH)-immunoreactive (IR) perikarya and fibers are widely distributed in the human hypothalamus. Since TH is the key and rate-limiting enzyme for catecholaminergic synthesis, these IR neurons may represent dopaminergic, noradrenergic or adrenergic neural elements. However, the distribution and morphology of these neurotransmitter systems in the human hypothalamus is not entirely known. Since the different catecholaminergic systems can be detected by identifying the neurons containing the specific key enzymes of catecholaminergic synthesis, in the present study we mapped the catecholaminergic elements in the human hypothalamus using immunohistochemistry against the catecholaminergic enzymes, TH, dopamine beta-hydroxylase (DBH) and phenylethanolamine-N-methyltransferase (PNMT). Only a few, PNMT-IR, adrenergic neuronal elements were found mainly in the infundibulum and the periventricular zone. DBH-IR structures were more widely distributed in the human hypothalamus occupying chiefly the infundibulum/infundibular nucleus, periventricular area, supraoptic and paraventricular nuclei. Dopaminergic elements were detected by utilizing double label immunohistochemistry. First, the DBH-IR elements were visualized; then the TH-IR structures, that lack DBH, were detected with a different chromogen. In our study, we conclude that all of the catecholaminergic perikarya and the majority of the catecholaminergic fibers represent dopaminergic neurons in the human hypothalamus. Due to the extremely small number of PNMT-IR, adrenergic structures in the human hypothalamus, the DBH-IR fibers represent almost exclusively noradrenergic neuronal processes. These findings suggest that the juxtapositions between the TH-IR and numerous peptidergic systems revealed by previous reports indicate mostly dopaminergic synapses.     

A direct catecholaminergic projection from the brainstem to the neurohypophysis of the rat

The presence of the catecholamines dopamine and noradrenaline in the posterior pituitary is well documented. Dopaminergic terminals are thought to derive from cells in the periventricular hypothalamus including the rostral arcuate nucleus, but the origin of the noradrenergic terminals is uncertain. The majority of central noradrenaline-containing neurons are in the brainstem and lesions of the ventral noradrenergic tract significantly reduce the noradrenaline content of the neural lobe. We have explored the possibility of a direct noradrenergic projection from the brainstem to the neural lobe using as a retrograde tract tracer horseradish peroxidase alone or in combination with immunohistochemical detection of tyrosine hydroxylase. Horseradish peroxidase was injected into the neural lobes of 20 rats using a ventral approach, and the animals were perfusion fixed 12 h later. In all 20 cases, cells retrogradely labelled with horseradish peroxidase were found not only in the periventricular zone and magnocellular nuclei of the hypothalamus but also in the A2 region of the brainstem. In all six cases simultaneously processed for horseradish peroxidase and tyrosine hydroxylase immunohistochemistry, retrogradely labelled cells in both the arcuate nucleus and A2 region were found to be tyrosine hydroxylase-positive. These findings demonstrate the presence of a direct projection from catecholaminergic neurons in the A2 region of the brainstem to the neurohypophysis. Since catecholaminergic neurons in this region are known to be noradrenergic and lesions of the ventral noradrenergic tract deplete the neurohypophysis of noradrenaline, these neurons may represent an important source of noradrenaline in the neurohypophysis.     

Studies on dopamine-, tyrosine hydroxylase- and aromatic L-amino acid decarboxylase-containing cells in the rat diencephalon: comparison between formaldehyde-induced histofluorescence and immunofluorescence

The morphology, number and distribution of catecholaminergic neurons, as visualized either with the aluminum-catalysed formaldehyde method for catecholamines or with the immunohistochemical method for the catecholamine-synthesizing enzymes tyrosine hydroxylase and aromatic L-amino acid decarboxylase, respectively, were analysed within the rat dorsal hypothalamus, ventral thalamus and adjoining regions (A11 and A13 cell groups). Both polyclonal rabbit and monoclonal mouse tyrosine hydroxylase antibodies were used in elution-restaining and double-staining experiments, respectively. Some of the animals also received spinal injections of the fluorescent tracer True Blue in order to retrogradely label cells projecting to the spinal cord. With respect to the number and distribution of catecholaminergic neurons in the A11 and medial A13 cell groups, including the spinal-projecting subpopulation, the results obtained with the two methods were very similar, indicating that within these regions of the CNS the two methods in principle visualize identical cell populations. However, the catecholaminergic cells were distinctly larger and their processes appeared more extensive with the immunohistochemical method. Animals processed for immunohistochemistry exhibited a lower total number of retrogradely labelled cells in the A11 area than those analysed with aldehyde-induced fluorescence despite the fact that both methods revealed similar numbers of retrogradely labelled tyrosine hydroxylase-positive and catecholamine-containing cells, respectively. The reason for these discrepancies, which are probably of methodological nature, are discussed. While this study shows that the results obtained with the two methods within the A11 and medial A13 cell group are very similar and thus strengthens the earlier proposed concept of the organization of the diencephalospinal dopaminergic system, it also documents that in intermingling and nearby CNS regions there are cell bodies which cannot be demonstrated with the aldehyde fluorescence method, but which still contain tyrosine hydroxylase and/or aromatic L-amino acid decarboxylase-like immunoreactivity. One explanation is low levels of enzyme and/or dopamine combined with a comparatively low sensitivity of the histochemical method. Thus, neurons containing both enzymes are probably dopaminergic, even if catecholamine fluorescence cannot be demonstrated. Neurons containing tyrosine hydroxylase, but lacking both aldehyde induced fluorescence and aromatic L-amino acid decarboxylase, may also still be dopaminergic.(ABSTRACT TRUNCATED AT 400 WORDS)     

Comparative distribution of NADPH-diaphorase activity and tyrosine hydroxylase immunoreactivity in the diencephalon and mesencephalon of the domestic chicken (Gallus domesticus)

We described the distribution of NADPH-diaphorase-containing neurons in relation to tyrosine hydroxylase immunoreactivity in the diencephalon and mesencephalon of the chicken. In the diencephalon, both markers were found in the lateral hypothalamus, dorsal hypothalamic area, hypothalamic periventricular nucleus, paraventricular nucleus and mamillary area. A close examination showed that the fine distribution of these markers differed slightly, so that they were never observed in the same neurons. In the mesencephalon, NADPH-diaphorase and tyrosine hydroxylase immunoreactivity were found in the ventral pedunculopontine area (nucleus tegmenti pedunculopontinus pars compacta, adjacent areas surrounding the quintofrontal tract and the nucleus mesencephalicus profundus ventralis), the coeruleus complex (locus coeruleus, ventral and dorsal subcoeruleus nuclei), the ventral tegmental area and the central gray. The majority of these neurons contained either diaphorase or tyrosine hydroxylase. Nevertheless, in a few cases both markers appeared to colocalize in the same neuron, typically in large perikarya of the ventral pedunculopontine area.     

Histamine-immunoreactive neurons in the brain of the teleost Gasterosteus aculeatus L. Correlation with hypothalamic tyrosine hydroxylase- and serotonin-immunoreactive neurons

The distribution of putative histaminergic neurons in the brain of a teleost, the three-spined stickleback, was investigated by means of immunocytochemistry using specific antibodies against histamine (HA), and conventional microscopy as well as confocal laser scanning microscopy. Histamine-immunoreactive (HAir) neurons form discrete populations ventral to the nucleus of the posterior recess (NRP) and in the nucleus saccus vasculosus (NSV), which belong to the periventricular hypothalamic nuclei. The neuronal somata are subependymally located, and do not possess apical neurites contacting the cerebrospinal fluid. They give rise to both long-range and local axonal projections. The local projections give rise to a field of dense punctate immunoreaction dorsal to the NRP and lateral to the NSV. Long-range projections are comprised of ascending projections to the thalamus, habenula, preoptic area and dorsal telencephalon; and descending projections via the posterior tuberal nucleus, ventrally to the nucleus interpeduncularis, and dorsally into the central gray. HAir neurons occur together with serotoninergic cerebrospinal fluid-contacting (CSFc) neurons in the NRP, and with tyrosine hydroxylase-immunoreactive (THir) neurons in the NSV. Although HAir elements occur together with THir ones in many brain areas, direct contacts between the two neurotransmitter systems are rare. The putative histaminergic neurons in the brain of the three-spined stickleback constitute a very discrete neuronal system, with a major projection area in the dorsal telencephalon in a region which is considered homologous with the dorsal pallium of land vertebrates.     

Stereological estimates of dopaminergic, GABAergic and glutamatergic neurons in the ventral tegmental area, substantia nigra and retrorubral field in the rat

Midbrain dopamine neurons in the ventral tegmental area, substantia nigra and retrorubral field play key roles in reward processing, learning and memory, and movement. Within these midbrain regions and admixed with the dopamine neurons, are also substantial populations of GABAergic neurons that regulate dopamine neuron activity and have projection targets similar to those of dopamine neurons. Additionally, there is a small group of putative glutamatergic neurons within the ventral tegmental area whose function remains unclear. Although dopamine neurons have been intensively studied and quantified, there is little quantitative information regarding the GABAergic and glutamatergic neurons. We therefore used unbiased stereological methods to estimate the number of dopaminergic, GABAergic and glutamatergic cells in these regions in the rat. Neurons were identified using a combination of immunohistochemistry (tyrosine hydroxylase) and in situ hybridization (glutamic acid decarboxylase mRNA and vesicular glutamate transporter 2 mRNA). In substantia nigra pars compacta 29% of cells were glutamic acid decarboxylase mRNA-positive, 58% in the retrorubral field and 35% in the ventral tegmental area. There were further differences in the relative sizes of the GABAergic populations in subnuclei of the ventral tegmental area. Thus, glutamic acid decarboxylase mRNA-positive neurons represented 12% of cells in the interfascicular nucleus, 30% in the parabrachial nucleus, and 45% in the parainterfascicular nucleus. Vesicular glutamate transporter 2 mRNA-positive neurons were present in the ventral tegmental area, but not substantia nigra or retrorubral field. They were mainly confined to the rostro-medial region of the ventral tegmental area, and represented approximately 2-3% of the total neurons counted ( approximately 1600 cells). These results demonstrate that GABAergic and glutamatergic neurons represent large proportions of the neurons in what are traditionally considered as dopamine nuclei and that there are considerable heterogeneities in the proportions of cell types in the different dopaminergic midbrain regions.     

Acute and repeated administration of fluphenazine-N-mustard alters levels of tyrosine hydroxylase mRNA in subsets of mesencephalic dopaminergic neurons.

Changes in striatal dopamine turnover and levels of tyrosine hydroxylase messenger RNA were examined in mice injected with D2 selective doses of fluphenazine-N-mustard, an irreversible blocker of dopaminergic receptors. The animals were killed at different times after acute and repeated injections of the drug and dopamine turnover was assessed by measuring dopamine and its metabolite, dihydroxyphenylalanine, in the striatum. Tyrosine hydroxylase mRNA was measured at the single-cell level in neurons of the substantia nigra pars compacta and the ventral tegmental area with quantitative in situ hybridization histochemistry. Acute treatment with fluphenazine-N-mustard induced an increase in both striatal dopamine turnover and the level of tyrosine hydroxylase mRNA in the substantia nigra but not the ventral tegmental area. After two days of repeated drug injections (twice daily), tyrosine hydroxylase mRNA was decreased in the substantia nigra despite the persistence of an elevated dopamine turnover in the striatum. The decrease in mRNA was still observed after four days of repeated treatment while, at that time, turnover values were not different from control. No changes were observed in the ventral tegmental area. The initial increase in tyrosine hydroxylase mRNA in substantia nigra pars compacta suggests that activation of nigrostriatal neurons triggers a very rapid increase in genomic expression of the enzyme. The following decrease in mRNA levels precedes desensitization to the effects of the drug on dopamine turnover, further illustrating a lack of correspondence between increased neurotransmission and levels of tyrosine hydroxylase mRNA in catecholaminergic neurons of the central nervous system.     

Distribution of catecholamines in the brain stem and spinal cord of the lizard Varanus exanthematicus: an immunohistochemical study based on the use of antibodies to tyrosine hydroxylase

Antibodies to tyrosine hydroxylase were used to study the distribution of nerve cells, fibers and terminals, containing catecholamines, in the lizard Varanus exanthematicus, by means of the indirect immunofluorescence technique. Tyrosine hydroxylase-containing cell bodies occurred in the hypothalamus, the ventral and dorsal tegmentum mesencephali, the substantia nigra, the isthmic reticular formation, in and ventrolaterally to the locus coeruleus, in the nucleus tractus solitarii and in a lateral part of the nucleus reticularis inferior. In addition tyrosine hydroxylase-containing cell bodies were found throughout the spinal cord, ventral to the central canal. Tyrosine hydroxylase-immunoreactive terminal areas in the brain stem were seen in the nucleus interstitialis of the fasciculus longitudinalis medialis, the nucleus raphes superior, the locus coeruleus, several parts of the reticular formation and the nucleus descendens nervi trigemini. Ascending catecholaminergic pathways could be traced from the ventral mesencephalic tegmentum as well as from the dorsal isthmic tegmentum rostralwards, through the lateral hypothalamus. These pathways correspond to the mesostriatal and isthmocortical projections respectively, as described in mammals. Furthermore, ascending catecholaminergic fibers could be traced from the catecholaminergic cell groups in the medulla oblongata to the isthmus, where they intermingle with the locus coeruleus neurons. These pathways correspond to the medullohypothalamic projection and to the dorsal periventricular system in mammals. Descending catecholaminergic fibers to the spinal cord pass via the dorsomedial part of the lateral funiculus, and mainly terminate in the dorsal horn. The results obtained in the present study have been placed in a comparative perspective, which illustrates the constancy of catecholaminergic innervation throughout phylogeny.     

Restricted co‐localization of glutamate and dopamine in neurons of the adult sea lamprey brain

Co‐localization of dopamine with other classical neurotransmitters in the same neuron is a common phenomenon in the brain of vertebrates. In mammals, some dopaminergic neurons of the ventral tegmental area and the hypothalamus have a glutamatergic co‐phenotype. However, information on the presence of this type of dopaminergic neurons in other vertebrate groups is very scant. Here, we aimed to provide new insights on the evolution of this neuronal co‐phenotype by studying the presence of a dual dopaminergic/glutamatergic neuron phenotype in the central nervous system of lampreys. Double immunofluorescence experiments for dopamine and glutamate in adult sea lampreys revealed co‐localization of both neurotransmitters in some neurons of the preoptic nucleus, the nucleus of the postoptic commissure, the dorsal hypothalamus and in cerebrospinal fluid‐contacting cells of the caudal rhombencephalon and rostral spinal cord. Moreover, co‐localization of dopamine and glutamate was found in dopaminergic fibres in a few brain regions including the lateral pallium, striatum, and the preoptic and postoptic areas but not in the brainstem. Our results suggest that the presence of neurons with a dopaminergic/glutamatergic co‐phenotype is a primitive character shared by jawless and jawed vertebrates. However, important differences in the distribution of these neurons and fibres were noted among the few vertebrates investigated to date. This study offers an anatomical basis for further work on the role of glutamate in dopaminergic neurons.     

Brain stem afferents to visual cortical areas 17, 18 and 19 in the cat, demonstrated by horseradish peroxidase

The origins of brain stem projections to the cytoarchitectonically different areas 17, 18 and 19 of the cat's visual cortex were studied following small horseradish peroxidase (HRP) injections. Labelled cells were counted in a dopaminergic nucleus (nucleus linearis rostralis (NLR)), other catecholaminergic nuclei (locus coeruleus, parabrachialis nuclei and nucleus subcoeruleus) and serotonergic nuclei (nucleus raphe dorsalis (NRD) and nucleus centralis superior (NCS)). Area 18 receives afferents from more locus coeruleus cells than either of areas 17 or 19. The number of labelled cells in the catecholaminergic nuclei far exceeds that in the serotonergic nuclei.     

A comparison of ventral tegmental neurons projecting to optic flow regions of the inferior olive vs. the hippocampal formation

The ventral tegmental area (catecholaminergic group A10) is a midbrain region characterized by concentrated dopaminergic immunoreactivity. Previous studies in pigeons show that the ventral tegmental area provides a robust projection to the hippocampal formation and to the medial column of the inferior olive. However, the distribution, morphology, and neurochemical content of the neurons that constitute these projections have not been resolved. In this study, we used a combination of retrograde tracing techniques and immunofluorohistochemistry to address these issues. Retrograde tracers were used to demonstrate that the distribution of ventral tegmental area neurons projecting to the hippocampus and the inferior olive overlap in the caudo-ventral ventral tegmental area. The hippocampus- and inferior olive-projecting ventral tegmental area neurons could not be distinguished based on morphology: most neurons had small- to medium-sized multipolar or fusiform soma. Double-labeling with fluorescent retrograde tracers revealed that the hippocampus- and medial column of the inferior olive-projecting neurons were found intermingled in the ventral tegmental area, but no cells were double labeled; i.e. individual ventral tegmental area neurons do not project to both the hippocampal formation and medial column of the inferior olive. Finally, we found that a minority (8.2%) of ventral tegmental area neurons providing input to the hippocampus were tyrosine hydroxylase-immunoreactive, whereas none of the inferior olive-projecting neurons were tyrosine hydroxylase positive. Combined, our findings show that the projections to the hippocampus and olivocerebellar pathway arise from intermixed subpopulations of ventral tegmental area neurons with indistinguishable morphology but only the hippocampal projection involves dopaminergic neurons. We suggest that equivalent projections from the ventral tegmental area to the hippocampal formation and inferior olive exist in mammals and discuss their potential role in the processing of optic flow and the analysis of self-motion.     

Ontogenesis of tyrosine hydroxylase-immunopositive structures in the rat hypothalamus. An atlas of neuronal cell bodies

The development of the catecholaminergic system in the hypothalamus and in the septal region was studied in rats from the 12th fetal day until the 9th postnatal day. Catecholaminergic structures were visualized with pre-embedding immunocytochemistry using antiserum to tyrosine hydroxylase. An intensification of diaminobenzidine product with silver and gold was additionally applied to make the immunocytochemical technique more sensitive. In this paper only the data on the appearance and distribution of the tyrosine hydroxylase-immunopositive neurons (cell bodies) are presented, whereas the catecholaminergic innervation of the hypothalamus with the tyrosine hydroxylase-immunopositive fibers is the topic of an accompanying paper. Sparse tyrosine hydroxylase-immunopositive neurons were first observed in the anlage of the hypothalamus and septal region on the 13th fetal day. Their number increased progressively with age and by the 15th fetal day they already gave rise to a large dorsal accumulation. From the 18th fetal day on, tyrosine hydroxylase immunopositive neurons began to occupy their definitive positions, mainly concentrating within the hypothalamus: in the zona incerta, periventricular and arcuate nuclei. To a lesser extent, they were concentrated in the medial preoptic area, suprachiasmatic, supraoptic, paraventricular, dorsomedial, and anterior hypothalamic nuclei. The data on the distribution of the tyrosine hydroxylase-immunopositive neurons both in the hypothalamus and in the septal region during ontogenesis are summarized in the precise atlas. Primarily small bi- and unipolar catecholaminergic neurons first observed in the youngest fetuses undergo cytodifferentiation during ontogenesis, giving rise to at least two different populations localized ventrally, mainly in the arcuate nucleus, and dorsally, in the zona incerta. The neurons of the former population remain similar to those of the youngest fetuses, whereas the neurons of the latter increase significantly in size, forming several long, highly ramified processes.     

Distribution and morphology of cholinergic, catecholaminergic and serotonergic neurons in the brain of Schreiber's long-fingered bat, Miniopterus schreibersii

The current study describes the nuclear parcellation and neuronal morphology of the cholinergic, catecholaminergic and serotonergic systems within the brain of a representative species of microbat. While these systems have been investigated in detail in the laboratory rat, and examined in several other mammalian species, no chiropterans, to the author's knowledge, have been examined. Using immunohistochemical stains for choline-acetyltransferase, tyrosine hydroxylase and serotonin, we were able to observe and document these systems in relation to the cytoarchitecture. The majority of cholinergic nuclei typically found in mammals were evident in the microbat, however we could not find evidence for choline-acetyltransferase immunopositive neurons in the Edinger–Westphal nucleus, parabigeminal nucleus, and the medullary tegmental field, as seen in several other mammalian species. A typically mammalian appearance of the catecholaminergic nuclei was observed, however, the anterior hypothalamic groups (A15 dorsal and ventral), the dorsal and dorsal caudal subdivisions of the ventral tegmental area (A10d and A10dc), and the ventral (pars reticulata) substantia nigra (A9v) were not present. The serotonergic nuclei were similar to that reported in all eutherian mammalian species studied to date. The overall complement of nuclei of these systems in the microbat, while different to the species examined in other orders of mammals, resembles most closely the complement seen in earlier studies of insectivore species, and is clearly distinguished from that seen in rodents, carnivores and primates. This data is discussed in terms of the phylogenetic relationships of the chiropterans.     

Ontogeny of GABA-immunoreactive neurons in the central nervous system in a teleost, Gasterosteus aculeatus L.

The inhibitory neurotransmitter γ-aminobutyric acid (GABA) is known to exert various neurotrophic actions in the developing nervous system, but little is known about its distribution in the central nervous system during early development. We have studied the development of GABA-immunoreactive (GABAir) neurons during embryogenesis of a teleost fish, the three-spined stickleback. As early as 51 h postfertilization (PF; hatching occurs 144–168 h PF, and the first monoaminergic neurons appear around 72 h PF) GABAir neurons appear in the ventral prosencephalon caudal to the optic recess, in the ventral meencephalon, and in the spinal cord. Then, there is a gradual addition of GABAir cell groups in the rostral prosencephalon and ventral rhombencephalon (66 h PF), dorsal and caudal hypothalamus and pretecturn (72 h PF), ventral hypothalamus (78 h PF), preoptic region, thalamus, and in the meencephalon and rhombencephalon (96 h PF). GABAir axons appear in the spinal cord already at 51 h PF, and then gradually appear in the various tracts of the early axonal scaffold of pathfinding fibers, so that by 96 h PF the entire axonal scaffold contains GABAir fibers. It appears likely that GABAergic axons contribute a major population to the formation of the axonal scaffold. Moreover, in the prosencephalon GABAir neurons are arranged in clusters that may reflect a neuromerec organization with six prosencephalic neuromeres.     

Distribution and morphology of cholinergic,catecholaminergic and serotonergic neurons in the brain of Schreiber's long-fingered bat,Miniopterus schreibersii

The current study describes the nuclear parcellation and neuronal morphology of the cholinergic, catecholaminergic and serotonergic systems within the brain of a representative species of microbat. While these systems have been investigated in detail in the laboratory rat, and examined in several other mammalian species, no chiropterans, to the author's knowledge, have been examined. Using immunohistochemical stains for choline-acetyltransferase, tyrosine hydroxylase and serotonin, we were able to observe and document these systems in relation to the cytoarchitecture. The majority of cholinergic nuclei typically found in mammals were evident in the microbat, however we could not find evidence for choline-acetyltransferase immunopositive neurons in the Edinger–Westphal nucleus, parabigeminal nucleus, and the medullary tegmental field, as seen in several other mammalian species. A typically mammalian appearance of the catecholaminergic nuclei was observed, however, the anterior hypothalamic groups (A15 dorsal and ventral), the dorsal and dorsal caudal subdivisions of the ventral tegmental area (A10d and A10dc), and the ventral (pars reticulata) substantia nigra (A9v) were not present. The serotonergic nuclei were similar to that reported in all eutherian mammalian species studied to date. The overall complement of nuclei of these systems in the microbat, while different to the species examined in other orders of mammals, resembles most closely the complement seen in earlier studies of insectivore species, and is clearly distinguished from that seen in rodents, carnivores and primates. This data is discussed in terms of the phylogenetic relationships of the chiropterans.     

Nuclear organization of some immunohistochemically identifiable neural systems in two species of the Euarchontoglires: A Lagomorph,Lepus capensis,and a Scandentia,Tupaia belangeri

The present study describes the organization of the nuclei of the cholinergic, catecholaminergic, serotonergic and orexinergic systems in the brains of two members of Euarchontoglires, Lepus capensis and Tupaia belangeri. The aim of the present study was to investigate the nuclear complement of these neural systems in comparison to previous studies on Euarchontoglires and generally with other mammalian species. Brains were coronally sectioned and immunohistochemically stained with antibodies against choline acetyltransferase, tyrosine hydroxylase, serotonin and orexin-A. The majority of nuclei revealed in the current study were similar between the species investigated and to mammals generally, but certain differences in the nuclear complement highlight potential phylogenetic interrelationships within the Euarchontoglires and across mammals. In the northern tree shrew the nucleus of the trapezoid body contained neurons immunoreactive to the choline acetyltransferase antibody with some of these neurons extending into the lamellae within the superior olivary nuclear complex (SON). The cholinergic nature of the neurons of this nucleus, and the extension of cholinergic neurons into the SON, has not been noted in any mammal studied to date. In addition, cholinergic neurons forming the medullary tegmental field were also present in the northern tree shrew. Regarding the catecholaminergic system, the cape hare presented with the rodent specific rostral dorsal midline medullary nucleus (C3), and the northern tree shrew lacked both the ventral and dorsal divisions of the anterior hypothalamic group (A15v and A15d). Both species were lacking the primate/megachiropteran specific compact portion of the locus coeruleus complex (A6c). The nuclei of the serotonergic and orexinergic systems of both species were similar to those seen across most Eutherian mammals. Our results lend support to the monophyly of the Glires, and more broadly suggest that the megachiropterans are more closely related to the primates than are any other members of Euarchontoglires studied to date.     

Hypophysiotrophic systems in the brain of the Atlantic salmon. Neuronal innervation of the pituitary and the origin of pituitary dopamine and nonapeptides identified by means of combined carbocyanine tract tracing and immunocytochemistry

The neuroanatomical organization of neurons projecting to the pituitary and the origin of pituitary dopamine and nonapeptides were investigated in the brain of the Atlantic salmon (Salmo salar). Carbocyanine tract tracing in combination with tyrosine hydroxylase, arginine vasotocin and isotocin immunocytochemistry for double labelling revealed a previously unknown organization of hypophysiotrophic cell groups and their extrahypothalamic projections, and provide the first direct identification in a teleost fish of the origin of the dopaminergic and nonapeptidergic innervation of the pituitary. The present data include identification of (1) hypophysiotrophic neurons in the ventral telencephalon and in the periventricular preoptic nucleus, (2) large (magnocellular) vasotocinergic hypophysiotrophic neurons in the most rostral extension of the preoptic area, (3) a distinct neuronal group located in a supraoptic/suprachiasmatic position in the anterior periventricular nucleus, that seems to be the major source of dopaminergic innervation of the pituitary, (4) the nonapeptidergic hypophysiotrophic neurons in the preoptic nucleus, (5) hypophysiotrophic neurons in the ventral and posterior hypothalamus of which some are of liquor-contacting type, (6) projections from hypophysiotrophic and non-hypophysiotrophic neurons in the preoptic nucleus to extrahypothalamic areas such as thalamic and periventricular pretectal nuclei, and (7) subdivisions within the preoptic nucleus that exhibit different combinations of hypophysiotrophic and extrahypothalamic efferent connections. Together with previous studies of retinohypothalamic projections and neurochemical organization of hypothalamic/preoptic areas, the present data suggest that the preoptic nucleus and the anterior periventricular nucleus in teleosts possess functional subdivisions with features that resemble those of the paraventricular, supraoptic and suprachiasmatic nuclei of other vertebrates. In the Atlantic salmon, specific dopaminergic and nonapeptidergic neuronal subdivisions are proposed to play a role for photoperiod control of endocrine activity.     

The differentiation of dopaminergic neurons in situ, in vivo, and in transplants

This article summarizes results obtained from studies on the differentiation of dopaminergic neurons in animal hypothalamus and human substantia nigra in situ, in vitro, and in transplants, as well as the role of the microenvironment in regulating this process. Four stages were identified in the differentiation of dopaminergic neurons from rat hypothalamus: a) formation of neurons from neuroepithelial precursor cells, b) expression of specific synthetic products (enzymes and dopamine itself) and mechanisms for transmembrane dopamine transport (reuptake and secretion in response to membrane depolarization), c) formation of permanent and transient efferent connections, and d) formation of afferent innervation and synaptogenesis. Along with dopaminergic neurons, rat fetuses contained neurons expressing only one of the dopamine-synthesizing enzymes and probably taking part in in situ dopamine synthesis. Differentiation of dopaminergic neurons was sexually dimorphic in terms of the dynamics of neuron formation and expression of enzymes involved in dopamine synthesis. A neurotransplantation model showed that humoral factors of placental and maternal origin had no significant effect on the differentiation of the dopaminergic neurons of the hypothalamus. As regards the dopaminergic neurons of the substantia nigra, expression of their specific phenotype in human fetuses started with the synthesis of tyrosine hydroxylase and co-maturation of the specific dopamine reuptake mechanism during the sixth week of development. During the next four weeks, specific uptake increased, and this appears to be a measure of the number of neurons and the growth of their processes. These data provide the basis for regarding the period from week 6 to week 10 as optimal for transplantation of dopaminergic neurons into the striatum of patients with Parkinson's disease. Suspensions of fetal substantia nigra cells enriched with dopaminergic neurons were introduced stereotaxically into a patient's striatum through a cannula. Positron emission tomography studies showed that the transplanted neurons survived within the host brain, underwent differentiation, and started to synthesize dopamine. The results of clinical assessment performed in parallel with these studies suggested that the transplanted dopaminergic neurons were involved in regulating striatal target neurons. Translated from Rossiiskii Fiziologicheskii Zhurmal imeni I. M. Sechenova, Vol. 84, No. 10, pp. 1019–1028, October, 1998.