Distribution of galanin-like immunoreactive elements in the brain of the adult lamprey Lampetra fluviatilis

Galanin is a brain-gut peptide present in the central nervous system of vertebrates and invertebrates. The distribution of galanin-like immunoreactive perikarya and fibers in the brain of the river lamprey Lampetra fluviatilis (Agnatha) has been studied immunocytochemically by using antisera against rat and porcine galanin. Galanin-like immunoreactive perikarya were seen in the telencephalon and mediobasal diencephalon. In the telencephalon, they were present in the nucleus olfactorius anterior, nucleus basalis, and especially, in the nucleus commissurae anterioris. The diencephalon contained most of the immunoreactive neurons. They were located in the nucleus commissurae praeinfundibularis, nucleus ventralis hypothalami, nucleus commissurae postinfundibularis, nucleus ventralis thalami, and nucleus dorsalis thalami pars medius. Most of the galanin-like immunoreactive infundibular neurons showed apical processes contacting the cerebrospinal fluid. Immunoreactive fibers and terminals were widely distributed throughout the neuraxis. In the telencephalon, the richest galaninergic innervation was found in the nucleus olfactorius anterior, lobus subhippocampalis, corpus striatum, and around the nucleus septi and the nucleus praeopticus. In the diencephalon, the highest density of galanin-like immunoreactive fibers was seen in the nucleus commissurae postopticae, nucleus commissurae praeinfundibularis, nucleus ventralis hypothalami, nucleus dorsalis hypothalami, and neurohypophysis. In the mesencephalon and rhombencephalon, the distribution of immunoreactive fibers was heterogeneous, being most pronounced in a region between the nucleus nervi oculomotorii and the nucleus interpeduncularis mesencephali, in the nucleus isthmi, and in the raphe region. A subependymal plexus of immunoreactive fibers was found throughout the ventricular system. The distribution of immunoreactive neurons and fibers was similar to that of teleosts but different to those of other vertebrate groups. The possible hypophysiotropic and neuroregulatory roles of galanin are discussed. © 1996 Wiley-Liss, Inc.     

Development of tyrosine hydroxylase-immunoreactive systems in the brain of the larval lamprey Lampetra fluviatilis

The development of the catecholaminergic system of the brain of the lamprey (Lampetra fluviatilis) was studied with immunocytochemistry in a series of larvae of different sizes by using two different antibodies directed against tyrosine hydroxylase (TH), the rate-limiting enzyme of catecholamine synthesis. In group 1 larvae (length: 29-54 mm, ages: 8 months to 1.5 years), the only TH-immunoreactive somata observed were located in the caudal wall of the recessus praeopticus (RP) and in the nucleus tuberculi posterioris (NTP). In group 2 larvae (length: 55-80 mm, ages: 1.5-2.5 years), the somata of immunolabeled cells of the NTP give rise to fibers, most of which are ascending and terminate in the corpus striatum. Additional immunoreactive cells are observed in the nucleus praeopticus (NP), which has differentiated, and in the spinal cord. In group 3 larvae (length: 81-110 mm, ages: 2.5-4 years), the spatial distribution of TH-immunoreactive elements (somata, fibers, and terminals) bears many resemblances to that seen in the adult. Immunolabeled cells may be observed in the olfactory bulb, in the nucleus commissurae postopticae (NCP), and in the nucleus dorsalis hypothalami (NDH). Nevertheless, some groups of TH-immunoreactive cells found in the adult are not observed in group 3 larvae; these may appear during the metamorphic phase. By comparative analysis, we show that, in spite of several differences, the spatiotemporal sequence of appearance of TH-immunoreactive cell bodies and fibers in the lamprey presents many similarities to that described in gnathostomes.     

Distribution of histaminergic neurons in the brain of the lamprey Lampetra fluviatilis as revealed by histamine-immunohistochemistry

An antiserum against conjugated histamine was used to study the distribution of histaminergic neurons in the CNS of the lamprey Lampetra fluviatilis. Numerous histamine-immunoreactive cell bodies were detected in the dorsal and ventral hypothalamic nuclei and in the adjacent postinfundibular commissural nucleus. Histamine-immunoreactive fibers of high density were present in the ventral hypothalamus, and fibers could also be traced dorsally from the hypothalamus to the corpus striatum and septal nucleus where they appeared to terminate in dense plexuses. Another, smaller group of histamine-immunoreactive perikarya was observed in the border area between mesencephalon and rhombencephalon, near the caudal pole of the mesencephalic reticular nucleus. Sparsely distributed histamine-immunoreactive fibers were present in the ventral mesencephalon. The distribution of histaminergic neurons in cyclostomes, which diverged very early from the main vertebrate line, shows similarities with the corresponding systems in the CNS of amphibians and mammals, which suggests that histaminergic neuronal systems are phylogenetically old and have been conserved during evolution.     

Calbindin-D28k immunoreactivity in the spinal cord of Xenopus laevis and its participation in ascending and descending projections

Immunohistochemistry for calbindin-D28k (CB) revealed that the spinal cord of Xenopus laevis possess a large number of CB-containing neurons widely distributed in both the dorsal and ventral horns, including areas which possess long ascending projections to supraspinal structures. In addition, the presence of CB-immunoreactive axons in the spinal funiculi suggested that descending projections containing this calcium binding protein may originate in different brainstem nuclei. Apart from mapping CB-containing elements in the spinal cord, a double labeling approach was used that combined the retrograde transport of dextran amines with CB immunohistochemistry. Thus, dextran amine injections into the lateral reticular region of the rhombencephalon, the parabrachial region, the mesencephalon and the dorsal thalamus revealed many retrogradely labeled cells in the spinal cord, a few number of which were double labeled for CB and found in the superficial dorsal horn and in the ventral medial region of the ventral horn. Their axons passed mainly via the lateral funiculus. Tracer application into the cervical spinal cord, combined with CB immunohistochemistry, resulted in retrogradely labeled cells throughout the brain, five groups of which showed CB immunoreactivity: (1) the mesencephalic trigeminal nucleus, (2) the laterodorsal tegmental nucleus, (3) the raphe nucleus, (4) the middle reticular nucleus and (5) the inferior reticular nucleus. The presence of CB in spinal pathways suggests that CB may play a role in controlling spinal cells, mainly subserving visceroceptive and nociceptive information to supraspinal levels, and might also modulate reticulospinal pathways.     

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.     

Morphology of the cells of origin of descending pathways to the spinal cord in Rana esculenta. A tracing study using cobaltic-lysine complex

The localization and morphological characteristics of neurons projecting to the spinal cord were studied with cobalt-filling technique. Cobaltic-lysine complex was iontophorized into descending pathways in the lumbar and cervical intumescence of the spinal cord and in some parts of the rhombencephalon. Well-filled cells located mainly in the rhombencephalon and telencephalon could be characterised as fusiform, triangular, multipolar and irregular neurons. Piriform and pyramidal cells predominated among projection neurons in the mesencephalon. Bilateral descending spinal pathways originate from the reticular nuclei of the rhombencephalon, nucleus vestibularis lateralis, nucl. anterodorsalis- and anteroventralis mesencephali, nucl. posterior thalami, nucl. ventralis hypothalami, area preoptica anterior and the striatum ventrale. Crossed pathways descend from the nucl. vestibularis descendens, nucl. tractus solitarii, nucl. cerebelli and the nucleus ruber. Uncrossed fibres originate from the nucl. tractus spinalis nervi trigemini, nucl. posteroventralis tegmenti, nucl. profundus mesencephali, nucl. fasciculi longitudinalis medialis and the nucleus ventrolateralis thalami. The organization of the frog's descending pathways is very similar to those in reptiles and in many ways to those in mammals. The possible synaptic connections of projection neurons have been discussed.     

Efferent connections of the vestibular nuclei in the rat: a neuromorphological study using PHA-L

The efferent connections of the superior, medial, lateral, and descending vestibular nuclei were studied with anterograde tracing methods in rats. The following areas of termination could be discerned: (1) In the diencephalon, labeled terminals were detected in the thalamus. (2) In the mesencephalon, the red nucleus and motor nuclei involved in eye movements were richly supplied by the vestibular nuclei. (3) In the rhombencephalon, extensive intrinsic connections of all vestibular nuclei were demonstrated. Strong commissural connections were found among the medial, superior, and descending vestibular nuclei. The inferior olive received labeled fibers exclusively from the lateral vestibular nuclei. Individual differences were demonstrated in the termination areas in the reticular formation. (4) In the spinal cord, most of the descending vestibular fibers were found in the ipsilateral anterior funiculus.     

Rostrocaudal distribution of 5-HT innervation in the lamprey spinal cord and differential effects of 5-HT on fictive locomotion

5-hydroxytryptamine (5-HT) is known to modulate the locomotion generator network in the lamprey spinal cord, but little is known about the pattern of 5-HT innervation along the spinal cord. The distribution of 5-HT-immunoreactive (5-HT-ir) cells and fibers, as well as the effects of 5-HT on the locomotor network in the rostral and caudal parts of the spinal cord were compared in two lamprey species, Lampetra fluviatilis and Petromyzon marinus. Intraspinal 5-HT cells form a very dense ventromedial plexus in which the dendrites of neurons forming the locomotor network are distributed. The number of 5-HT cells and varicosities in this plexus decreases in the fin area (segments 70–90), and then increases somewhat in the most caudal segments. The descending 5-HT fibers from the rhombencephalon are located in the lateral and ventral columns, and their numbers gradually decrease to around 50% in the tail part of the spinal cord. In contrast, the number of 5-HT-ir axons in the dorsal column remains the same along the spinal cord. Bath application of both N-methyl-D-aspartic acid (NMDA; 20–250 μM) and D-glutamate (250–1000 μM) was used to induce fictive locomotion in the isolated spinal cord. Bath application of 5-HT (1 μM) reduced the burst frequency in the presence of NMDA. The 5-HT effect was, however, significantly greater in the rostral as compared to the caudal part. With D-glutamate, the 5-HT effect was instead more pronounced in the caudal spinal cord. To account for this difference in 5-HT effects on NMDA- and D-glutamate-induced fictive locomotion, the cellular effect of D-glutamate was further investigated. It activates not only NMDA, but also alpha amino-3-hydroxy-5-methyl-4-isoxyl propionate (AMPA)/kainate and metabotropic glutamate receptors. In contrast to NMDA, D-glutamate did not elicit tetrodotoxin (TTX)-resistant membrane potential oscillations. This difference in action between NMDA (selective NMDA receptor agonist) and D-glutamate (mixed agonist) may partially account for the differences in effect of 5-HT on the locomotor pattern. © Wiley-Liss, Inc.     

Cells of origin of pathways descending to the spinal cord in two chondrichthyans,the shark Scyliorhinus canicula and the ray Raja clavata

The cells of origin of pathways descending to the spinal cord in the shark Scyliorhinus canicula and in the ray Raja clavata have been demonstrated by using the horseradish peroxidase (HRP) technique. Following HRP injections in the spinal cord of Scyliorhinus (fourth to sixth segment) and of Raja (15th to 20th segment) labeled neurons could be identified in the rhombencephalon, the mesencephalon, and in the diencephalon. Cells of origin of diencephalic nuclei, which project to the spinal cord, were observed in the nucleus periventricularis hypothalami and in the thalamus ventralis pars medialis which can in this respect be considered hypothalamic. Descending pathways from mesencephalic structures originate from the interstitial nucleus of the fasciculus longitudinalis medialis, the tectum mesencephali, the nucleus intercollicularis, the tectotegmental junction zone, and from diffusely arranged tegmental neurons. A contralateral rubrospinal pathway could be recognized in Raja, but not in Scyliorhinus. Rhombencephalic cells of origin of pathways descending to the spinal cord were found in all parts of the reticular formation, i.e., the nucleus raphes inferior, the nucleus reticularis inferior, medius, superior, and isthmi, in two vestibular nuclei, and in three nuclei, which have been tentatively indicated as nucleus B, F, and G. Furthermore cells of origin of descending pathways have been found in the nucleus tractus descendens nervi trigemini, in the nucleus funiculi lateralis, and in the nucleus tractus solitarii. The descending pathways of the two species studied have been compared with those of other vertebrates. It is concluded that the basic pattern in the organization of descending pathways to the spinal cord, as proposed by ten Donkelaar ('76) for terrestrial vertebrates, also holds for cartilaginous fishes.     

Organization of the catecholaminergic systems in two basal actinopterygian fishes,Polypterus senegalus and Erpetoichthys calabaricus (Actinopterygii: Cladistia)

Cladistians are a group of basal nonteleost actinopterygian fishes that represent an interesting group for the study of primitive brain features, most likely present in the ancestral Osteichthyes. We have investigated the catecholaminergic (CA) systems in the brain of two representative cladistian species, the bichir Polypterus senegalus and the reedfish Erpetoichthys calabaricus, by means of antibodies against tyrosine hydroxylase (TH; the first enzyme in the synthesis of catecholamines) and dopamine (DA). Double immunohistofluorescence was performed for simultaneous detection of TH with nitric oxide synthase, choline acetyltransferase, calbindin, calretinin, and serotonin, aiming to accurately establish the localization of the CA neurons and to assess possible interactions between these neuroactive substances. All forebrain CA groups of cladistians are dopaminergic, whereas noradrenergic cells are located within the rhombencephalon. Distinct groups of DA immunoreactive (DA-ir) cells were observed in the olfactory bulb, subpallium, and preoptic area of the telencephalon. Hypothalamic groups were detected in the suprachiasmatic nucleus, retrotuberal and retromamillary areas and, in particular, the paraventricular organ showed immunoreactivity to dopamine but not to TH. Diencephalic DA-ir groups were detected in the prethalamus, posterior tubercle, and pretectum. A small DA-ir cell population was observed in the midbrain tegmentum only in Polypterus. CA cell groups were also located in the locus coeruleus, solitary tract nucleus, and area postrema within the rhombencephalon, the spinal cord, and the retina. The comparison of these results with other vertebrates, using a neuromeric analysis, shows highly conserved traits in all vertebrates studied but also evidences particular characteristics of actinopterygian fishes.     

Distribution of substance P-like immunoreactivity in the brain of the newt (Triturus cristatus)

The distribution of immunoreactive substance P (sP)-containing structures in the newt brain and spinal cord was explored with an indirect im-munofluorescence method. Five sP-positive elements were detected: perikarya, dots, fibers, pericellular appositions, and pipe-shaped structures. Perikarya were seen at the levels of the spinal ganglia, spinal cord, raphe nucleus, interpeduncular nucleus, mesencephalon, preoptic area, infundibulum, dorsocaudal part of the ventral hypothalamus, habenula, and corpus striatum. Pericellular terminals were observed in periventricular areas, known to be rich in catecholaminergic cells; pipe-shaped structures were ob-served from the corpus striatum to diencephalon, and in mesencephalon. The olfactory nerve and nuclei were devoid of sP-positive elements. Six sP-immunofluorescent pathways were detected. One of them is composed of axons with huge varicosities and extends from the lateral spinal cord area to the mesencephalon. This pathway has not been described as yet in other animals and could be peculiar to the newt.     

The mormyrid brainstem. I. Distribution of brainstem neurones projecting to the spinal cord in Gnathonemus petersii. An HRP study

The sources of the descending spinal tracts were identified in the teleost fish Gnathonemus petersii by retrograde HRP transport. HRP injections were made at two spinal levels, either at level of the caudal end of the dorsal fin, anterior to the electric organ, or at the pectoral fin. In both cases all labeled cells were found in the rhombencephalon and the mesencephalic tegmentum. No labeled cells were observed either in the cerebellum and lateral line lobes or in the dorsal mesencephalon i.e. torus semicircularis and mesencephalic tectum or in the telencephalon. Following caudal spinal injections, the majority of the labeled cells were grouped in a median and a ventrolateral column of the rhombencephalic reticular formation. The latter is composed of three parts corresponding to the nucleus reticularis inferior, medius and superior. Both Mauthner cells, all the cells in the medullary relay nucleus controlling the electric organ discharge and a few cells in the posterior part of the magnocellular octaval nucleus were labeled. In the mesencephalon, four nuclei were identified by HRP labeling: the nucleus of the medial longitudinal fasciculus, the nucleus reticularis mesencephali and the anterior and posterior tegmental mesodiencephalic nuclei. The rostral injections revealed several additional spinal projections from the descending vestibular and tangential nuclei, from the medial part of the magnocellular nucleus and, finally, from the rostral periventricular gray of the mesencephalon. Also, after such injections, a greater number of cells were labeled in the reticular formation, especially in the median column and in the inferior reticular nucleus. The results suggest that the rostral spinal cord has a larger connection with the acoustico-vestibular area and the medullary reticular formation than the caudal spinal cord. In contrast, the mesencephalic nuclei, probably linked to the mesencephalic tectum and the pretectal area, appears to be a coordinating apparatus between the visual system and the trunk/tail musculature. Thus, it appears that teleost fish possess the same basic equipment of descending spinal pathways as higher vertebrates.     

Distribution of melanin-concentrating hormone-like immunoreactivity in the central nervous system of Rana esculenta

The distribution of salmon and rat melanin-concentrating hormone (MCH)-like and neuropeptide glutamate-isoleucine (NEI)-like immunoreactivity in the brain and spinal cord of the frog Rana esculenta was studied with immunohistochemistry. In the telencephalon, only fibers showed immunoreactivity in the olfactory bulb, lateral pallium, diagonal band, septum, and the amygdala. Immunoreactive fibers were abundant in all diencephalic structures, except the optic tract, the visual neuropils, and the habenula. Several cells in the central thalamic nucleus and a few in the suprachiasmatic nucleus were stained with the MCH antisera. Cells and their processes were intensely stained in the dorsal hypothalamus with the MCH and NEI antisera. Immunoreactive fibers were found in all tegmental nuclei and the white matter of the mesencephalon. They formed terminal plexuses in the deep layers of the optic tectum and the laminar nucleus of the torus semicircularis. Immunoreactive fibers were sparse in the rhombencephalon and the spinal cord.     

Spinal cord development in anuran larvae: II. Ascending and descending pathways

The ontogeny of ascending and descending spinal pathways was examined in bullfrog (Rana catesbeiana) tadpoles using the transported histochemical marker, horseradish peroxidase (HRP). The adult pattern of brainstem projections to lumbar spinal cord is evident as early as larval stage I (Taylor and Kollros, Anat. Rec., 94:7–24, 1946), although the number and size of projecting cells increases as the animal matures. These projections arise from presumptive hypothalamic neurons at the diencephalicmesencephalic border as well as from neurons of the vestibular nucleus, oculomotor nucleus, and reticular formation. In contrast to the stability of the pattern of descending projections, the sources of fibers ascending to the brainstem change during larval life. In early larval stages, brainstem projections from lumbar spinal cord arise primarily from Rohon-Beard cells and neurons of the superficial dorsal horn. In later stages, neurons in the intermediate and ventral areas of the spinal gray can also be retrogradely labeled by HRP application to the brainstem at the level of the VIIIth nerve. Evidence of the existence of dorsal column and lateral cervical nuclei in adult frog and tadpoles older than stage VIII is presented. The ascending projections of embryonically born primary neurons were also investigated. Rohon-Beard cells, which are sensory neurons with their cell bodies in the spinal cord, were found to send ascending processes as least as far rostral as the level of the VIIIth nerve entry zone. Anterolateral and dorsal marginal cells, probable homologs, respectively, of mammalian spinal border cells and cells of Waldeyer (1888), were also found to project rostrally at least to the rhombencephalon. These marginal cells persisted through metamorphosis into adulthood.     

Immunohistochemical studies of cholecystokininlike peptides and their relation to 5-HT, CGRP, and bombesin immunoreactivities in the brainstem and spinal cord of lampreys

The distribution of cholecystokinin (CCK)-like immunoreactivity in the brainstem and spinal cord of lampreys was studied by using CCK antisera with different properties. In the spinal cord, three separate systems reacted with CCK antisera: (1) A ventral and lateral fiber system descending from a group of neurons in the posterior reticular nucleus of the rhombencephalon was labeled by both a C-terminal-directed CCK antiserum and a monoclonal CCK antibody. (2) A dorsal root-dorsal column system of fibers originating from cell bodies in the dorsal root ganglia was labeled only by the C-terminal CCK antiserum. This CCK immunoreactivity could be abolished by preabsorption with calcitonin-gene-related peptide (CGRP), suggesting that it was due to cross-reactivity with a CGRP-like peptide. This system also contained 5-hydroxytryptamine (5-HT)-, bombesin-, and CGRP-like immunoreactivities. (3) An intraspinal system of 5-HT neurons was labeled with an antiserum to the midportion of CCK-33 but not by the other CCK antisera. The CCK labeling of this system was difficult to reduce by preabsorption with CCK peptide and thus appeared to be nonspecific. Groups of cell bodies in the middle reticular nucleus of the rhombencephalon, the reticular nucleus of the mesencephalon, and the hypothalamus were labeled by both the C-terminal and the monoclonal CCK antisera. The gut contained two types of CCK-like immunoreactivity, one of which appeared to be due to cross-reactivity with CGRP. A biochemical analysis showed that the content of CCK was low in the spinal cord compared to the brain, and these results agreed with the immunohistochemical findings.     

Distribution of thyrotropin-releasing hormone (TRH) immunoreactivity in the brain of the zebrafish (Danio rerio)

The distribution of thyrotropin-releasing hormone (TRH) in the brain of the adult zebrafish was studied with immunohistochemical techniques. In the telencephalon, abundant TRH-immunoreactive (TRHir) neurons were observed in the central, ventral, and supra- and postcommissural regions of the ventral telencephalic area. In the diencephalon, TRHir neurons were observed in the anterior parvocellular preoptic nucleus, the suprachiasmatic nucleus, the lateral hypothalamic nucleus, the rostral parts of the anterior tuberal nucleus and torus lateralis, and the posterior tuberal nucleus. Some TRHir neurons were also observed in the central posterior thalamic nucleus and in the habenula. The mesencephalon contained TRHir cells in the rostrodorsal tegmentum, the Edinger-Westphal nucleus, the torus semicircularis, and the nucleus of the lateral lemniscus. Further TRHir neurons were observed in the interpeduncular nucleus. In the rhombencephalon, TRHir cells were observed in the nucleus isthmi and the locus coeruleus, rostrally, and in the vagal lobe and vagal motor nucleus, caudally. In the forebrain, TRHir fibers were abundant in several regions, including the medial and caudodorsal parts of the dorsal telencephalic area, the ventral and commissural parts of the ventral telencephalic area, the preoptic area, the posterior tubercle, the anterior tuberal nucleus, and the posterior hypothalamic lobe. The dorsal thalamus exhibited moderate TRHir innervation. In the mesencephalon, the optic tectum received a rich TRHir innervation between the periventricular gray zone and the stratum griseum centrale. A conspicuous TRHir longitudinal tract traversed the tegmentum and extended to the rhombencephalon. The medial and lateral mesencephalic reticular areas and the interpeduncular nucleus were richly innervated by TRHir fibers. In the rhombencephalon, the secondary gustatory nucleus received abundant TRHir fibers. TRHir fibers moderately innervated the ventrolateral and ventromedial reticular area and richly innervated the vagal lobe and Cajal's commissural nucleus. Some TRHir fibers coursed in the lateral funiculus of the spinal cord. Some TRHir amacrine cells were observed in the retina. The wide distribution of TRHir neurons and fibers observed in the zebrafish brain suggests that TRH plays different roles. These results in the adult zebrafish reveal a number of differences with respect to the TRHir systems reported in other adult teleosts but were similar to those found during late developmental stages of trout (Díaz et al., 2001).     

Brainstem neurons projecting to different levels of the spinal cord of the dogfish Scyliorhinus canicula.

Neurons in the brain that project to different levels of the spinal cord in the dogfish Scyliorhinus canicula have been identified by retrograde labelling with horseradish peroxidase. Injections have been made at four different levels, namely, cervical (segments 3-6), pectoral (segments 16-18), pelvic (segments 34-36) and caudal (segments 60-80). Labelled neurons were located in the diencephalon, mesencephalon and rhombencephalon. Twenty nuclei contained labelled cells following cervical injections. Fourteen of these contained labelled cells following pectoral injections as did nine following pelvic injections and only seven after caudal injections. Of the three diencephalic nuclei projecting to the cord only nucleus thalamus ventralis pars medialis projected further than cervical regions. From the mesencephalon, the tectospinal projection, which arises from tectal and tectotegmental regions, reaches only as far as the cervical cord. The reticulospinal system, arising from cell groups in both the mesencephalon and rhombencephalon, provides the major descending pathway and reaches the most caudal levels. From the octaval region a vestibulospinal pathway arises from nucleus octavus magnocellularis, to reach all levels of the cord, and from nucleus octavus descendens, to project only as far as the pectoral cord. Other rhombencephalic cell groups--the trigeminal nuclei and nuclei B, F and G--project at least as far as the pectoral cord. We conclude that few brainstem nuclei can directly affect the whole spinal cord, whereas many nuclei may have an impact on its most rostral regions; here we have found labelled spinal interneurons with long descending axons that reach the most caudal cord levels.     

Organization of the corticotropin-releasing hormone and corticotropin-releasing hormone-binding protein systems in the central nervous system of the sea lamprey Petromyzon marinus

The expression of the corticotropin-releasing hormone (PmCRH) and the CRH-binding protein (PmCRHBP) mRNAs was studied by in situ hybridization in the brain of prolarvae, larvae, and adults of the sea lamprey Petromyzon marinus. We also generated an antibody against the PmCRH mature peptide to study the distribution of PmCRH-immunoreactive cells and fibers. PmCRH immunohistochemistry was combined with antityrosine hydroxylase immunohistochemistry, PmCRHBP in situ hybridization, or neurobiotin transport from the spinal cord. The most numerous PmCRH-expressing cells were observed in the magnocellular preoptic nucleus-paraventricular nucleus and in the superior and medial rhombencephalic reticular formation. PmCRH expression was more extended in adults than in larvae, and some cell populations were mainly (olfactory bulb) or only (striatum, ventral hypothalamus, prethalamus) observed in adults. The preopto-paraventricular fibers form conspicuous tracts coursing toward the neurohypophysis, but many immunoreactive fibers were also observed coursing in many other brain regions. Brain descending fibers in the spinal cord mainly come from cells located in the isthmus and in the medial rhombencephalic reticular nucleus. The distribution of PmCRHBP-expressing neurons was different from that of PmCRH cells, with cells mainly present in the septum, striatum, preoptic region, tuberal hypothalamus, pretectum, pineal complex, isthmus, reticular formation, and spinal cord. Again, expression in adults was more extended than in larvae. PmCRH- and PmCRHBP-expressing cells are different, excluding colocalization of these substances in the same neuron. Present findings reveal a complex CRH/CRHBP system in the brain of the oldest extant vertebrate group, the agnathans, which shows similarities but important divergences with that of mammals.     

Alpha-melanocyte stimulating hormone immunoreactivity in the brain of the cichlid fish Oreochromis mossambicus

This study reports the distribution of a pro-opiomelanocortin-derived neuropeptide α–MSH in the brain of the cichlid fish Oreochromis mossambicus. α–MSH-ir fibres were found in the granule cell layer of the olfactory bulb, the medial olfactory tract, the pallium and the subpallium, whereas in the preoptic area of the telencephalon, few large α–MSH-ir perikarya along with extensively labeled fibres were observed close to the ventricular border. Dense network of α–MSH-ir fibres were seen in the hypothalamic areas such as the nucleus preopticus pars magnocellularis, the nucleus preopticus pars parvocellularis, the suprachiasmatic nucleus, the nucleus anterior tuberis, the paraventricular organ, the subdivisions of the nucleus recessus lateralis and the nucleus recessus posterioris. In the nucleus lateralis pars medialis, some α–MSH-ir perikarya and fibres were found along the ventricular margin. In the diencephalon, numerous α–MSH-ir fibres were detected in the nucleus posterior tuberis, the nucleus of the fasciculus longitudinalis medialis and the nucleus preglomerulosus medialis, whereas in the mesencephalon, α–MSH-ir fibres were located in the optic tectum, the torus semicircularis and the tegmentum. In the rhombencephalon, α–MSH-ir fibres were confined to the medial octavolateralis nucleus and the descending octaval nucleus. In the pituitary gland, densely packed α–MSH-ir cells were observed in the pars intermedia region. The widespread distribution of α–MSH-immunoreactivity throughout the brain and the pituitary gland suggests a role for α–MSH peptide in regulation of several neuroendocrine and sensorimotor functions as well as darkening of pigmentation in the tilapia.