Organization of the pallium in the fire-bellied toad Bombina orientalis. I: Morphology and axonal projection pattern of neurons revealed by intracellular biocytin labeling

The cytoarchitecture and axonal projection pattern of pallial areas was studied in the fire-bellied toad Bombina orientalis by intracellular injection of biocytin into a total of 326 neurons forming 204 clusters. Five pallial regions were identified, differing in morphology and projection pattern of neurons. The rostral pallium receiving the bulk of dorsal thalamic afferents has reciprocal connections with all other pallial areas and projects to the septum, nucleus accumbens, and anterior dorsal striatum. The medial pallium projects bilaterally to the medial pallium, septum, nucleus accumbens, mediocentral amygdala, and hypothalamus and ipsilaterally to the rostral, dorsal, and lateral pallium. The ventral part of the medial pallium is distinguished by efferents to the eminentia thalami and the absence of contralateral projections. The dorsal pallium has only ipsilateral projections running to the rostral, medial, and lateral pallium; septum; nucleus accumbens; and eminentia thalami. The lateral pallium has ipsilateral projections to the olfactory bulbs and to the rostral, medial, dorsal, and ventral pallium. The ventral pallium including the striatopallial transition area (SPTA) has ipsilateral projections to the olfactory bulbs, rostral and lateral pallium, dorsal striatopallidum, vomeronasal amygdala, and hypothalamus. The medial pallium can be tentatively homologized with the mammalian hippocampal formation, the dorsal pallium with allocortical areas, the lateral pallium rostrally with the piriform and caudally with the entorhinal cortex, the ventral pallium with the accessory olfactory amygdala. The rostral pallium, with its projections to the dorsal and ventral striatopallidum, resembles the mammalian frontal cortex.     

Afferent and efferent connections of the bullfrog medial pallium.

Horseradish peroxidase or tritiated proline was unilaterally injected into the medial pallium in bullfrogs in order to determine the sources of afferent projections to the medial pallium and the targets of pallial efferent projections. Some cells in all telencephalic centers, except the corpus striatum and the pars lateralis of the amygdala, project to the ipsilateral medial pallium. The medial pallium receives projections from fewer centers in the contralateral hemisphere, which include the medial septal nucleus, the pars medialis of the amygdala, the bed nucleus of the pallial commissure and the medial pallium. The raphe nucleus and the anterior thalamic nuclei appear to be the only sources of afferents to the medial pallium from outside the telencephalon. Efferents of the medial pallium are far more extensive than reported in earlier studies. The medial pallium projects ipsilaterally to all telencephalic nuclei, with the exception of a large part of the corpus striatum, and contralaterally to the medial septal nucleus, the olfactory tubercle, amygdala, medial pallium and bed nucleus of the pallial commissure. Extensive efferent projections also terminate in preoptic and hypothalamic regions, as well as in most thalamic relay nuclei, the pretectum and, possibly, the optic tectum. Similarities to the medial pallium in other tetrapods and to that in mammals suggest that the medial pallium in anurans is homologous to the subicular and CA fields and, possibly, the dentate gyrus in mammals. However, the extensive projections of the medial pallium to the dorsal thalamus and pretectum in anurans may be primitive features of the medial pallium retained in anurans, or uniquely derived features in anurans.     

Morphology, axonal projection pattern, and responses to optic nerve stimulation of thalamic neurons in the salamander Plethodon jordani

In the salamander Plethodon jordani, the morphology and axonal projections of thalamic (TH) neurons and their responses to electrical optic nerve stimulation were determined by intracellular recording and biocytin labeling under in vitro, whole-brain conditions. Based on their axonal projections, labeled neurons (n = 76) were divided into the following groups: TH1 neurons, with mostly ipsilateral projections to the striatum; TH2 neurons, with ipsilateral or bilateral projections to the medial amygdala and nucleus accumbens; TH3 neurons, with bilateral projections to the medial and dorsal pallium; TH4 neurons, with mostly ipsilateral projections to the striatum and ipsilateral projections to the tectum opticum, tegmentum, and rostral medulla oblongata; and TH5 neurons, with ipsilateral projections to the tegmentum, medulla oblongata, and rostral spinal cord without (TH5.1) or with (TH5.2) additional projections to the optic tectum. TH1-TH4 neurons are found in the dorsal thalamus and around the sulcus medialis, and TH5 neurons are found in the ventral thalamus. Labeled neurons with ascending projections, i.e., the more dorsally situated TH1-TH4 neurons, are mostly inhibited by electrical stimulation of the optic nerve and have significantly longer latencies (mean +/- S.D., 42.1 +/- 11.6 msec) than neurons with exclusively descending projections, i.e., the ventrally located TH5 neurons (8.5 +/- 6.1 msec), which receive the bulk of retinal afferents and show excitation at electrical optic nerve stimulation. Neurons recorded without labeling in the dorsal thalamus likewise exhibit mostly inhibition and have significantly longer latencies (35.7 +/- 18.9 msec) than those recorded in the ventral thalamus (10.9 +/- 7.7 msec), which mostly show excitation. None of the neurons recorded in the dorsal thalamus followed repetitive stimulation of the optic nerve. Thus, neurons situated in the dorsal thalamus and projecting to pallial or subpallial telencephalic targets are unlikely to receive monosynaptic or oligosynaptic, excitatory retinal input. Accordingly, no retino-thalamo-telencephalic pathway homologous to that found in amniotes appears to exist in salamanders.     

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

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

Immunohistochemical study of the telencephalon of the spiny dogfish, Squalus acanthias

The paucity of experimental data and the differences in telencephalic organization between sharks and other jawed vertebrates have complicated telencephalic comparisons. The distribution of neuropeptides has been extremely useful in recognizing and comparing major subdivisions of the telencephalon among vertebrates. Immunohistochemical techniques were therefore used to study the distribution of substance P (SP), leucin-enkephalin (LENK), and serotonin (5HT), as well as tyrosine hydroxylase (TH), an indicator of catecholamines, in the telencephalon of the spiny dogfish. The distribution of SP and LENK provides a clear distinction between pallial and subpallial portions of the telencephalon. Two regions of the ventrolateral telencephalon, area superficialis basalis and area periventricularis ventrolateralis, exhibit histochemical similarities to the pallidal and striatal subdivisions, respectively, of the basal ganglia in amniotes. Lower densities of LENK+ and SP+ perikarya and fibers occur in the medial pallium and the pars centralis of the dorsal pallium. Similar histochemical traits characterize the sensory thalamorecipient telencephalic structures in amniotes. The lateral pallium in dogfishes is distinguished by the presence of large numbers of TH+ neurons with radially oriented processes. The presence of these distinctive cells also in the medial wall of the rostral telencephalon suggests that the lateral pallium has a medial extension that is situated ventral to the medial pallium. Neurons containing TH were widely distributed in the telencephalon of spiny dogfish and were particularly abundant in the dorsal pallium, olfactory pallium, and area superficialis basalis. It is currently unclear whether these TH+ telencephalic neurons are, in fact, catecholaminergic or merely contain a TH-like substance unrelated to catecholamine synthesis.     

Afferent connections of septal nuclei of the domestic chick (Gallus domesticus): a retrograde pathway tracing study

The afferents to the septum of the domestic chicken were studied using retrograde tracers, rhodamine conjugated latex bead or Fast Blue, placed in different septal subregions. The results were verified by anterograde tracer injections deposited to selected areas. The main telencephalic afferents to the septum arise ipsilaterally from the hippocampal formation, dorsolateral corticoid area, piriform cortex, amygdaloid pallium, and the ventral pallidum. Contralateral afferents originate from the lateral septum and the amygdaloid pallium. A massive bilateral projection arises from the lateral hypothalamus. Other hypothalamic afferents arise from the periventricular, paraventricular and anterior medial nuclei, and the premammillary and mammillary areas. The dorsal thalamic nuclei (dorsal medial anterior and posterior) and the reticular dorsal nuclei also contribute septal afferents. Brainstem afferents arise bilaterally from the ventral tegmental area, substantia nigra, central gray, A8, locus coeruleus, ventral subcoeruleus nucleus, and raphe nuclei. The main terminal fields for septal afferents lie in the lateral septal nucleus and the belt of medial septal nucleus. The core of the latter is invaded mainly by fibers from the brainstem, presumably belonging to the ascending activating system. The septal afferents of the chicken are largely similar to those of other avian and nonavian species. The most prominent differences with previous pigeon data were found in the subregional selectivity of the hippocampal formation, dorsolateral corticoid area, mammillary nuclei, some dorsal thalamic nuclei, substantia nigra, and subcoeruleus nuclei in their projections to defined septal nuclei.     

Telencephalic efferents of the tiger salamander Ambystoma tigrinurn tigrinum (Green)

The efferent projections of the telencephalon in the tiger salamander were examined by the Nauta and Fink-Heimer methods following unilateral hemispherectomies, rostral hemispheric ablations and pallial lesions. The cerebral hemisphere connects with most areas of the contralateral hemisphere via the pallial, anterior and habenular commissures. The descending fibers travel in the medial and lateral forebrain bundles and in the tracts comprising the stria medullaris. Degenerating fibers and terminals were present throughout the diencephalon but were more abundant ipsilaterally. Fibers reach the pretectum and optic tectum via dorsal and ventral pathways. There is a heavy projection to the midbrain tegmentum and a sparse projection to the tectum via the ipsilateral lateral forebrain bundle. This tract continues into the medulla oblongata and the cervical spinal cord. Rostral and dorsal hemispheric ablations revealed that the majority of fibers forming the olfacto-peduncular tract originate in the ventral, rostral one-third of the hemisphere. It was also determined that the majority of the descending efferent fibers located in the lateral forebrain bundle originate from the caudal lateral hemispheric wall, and that these fibers form connections characteristic of mammalian corticofugal and striatofugal systems. The cytoarchitecture and connections of the caudal lateral hemispheric wall suggest that it is homologous to parts of motor isocortex and amygdala of amniotes.     

Thalamo-telencephalic connections: new insights on the cortical organization in reptiles

Tracer injections into the dorsal tier of the lacertilian dorsal thalamus revealed an extensive innervation of the cerebral cortex. The medial cortex, the dorsomedial cortex, and the medial part of the dorsal cortex received a bilateral projection, whereas the lateral part of dorsal cortex and the dorsal part of the lateral cortex received only an ipsilateral thalamic projection. Thalamocortical fibers were found superficially in all cortical regions, but in the dorsal part of the lateral cortex, varicose axons within the cellular layer were also observed. The bilateral thalamocortical projection originates from a cell population located throughout the dorsolateral anterior nucleus, whereas the ipsilateral input originates mainly from a rostral neuronal subpopulation of the nucleus. This feature suggests that the dorsolateral anterior nucleus consists of various parts with different projections. The dorsal subdivision of the lateral cortex displayed hodological and topological (radial glia processes) features of a dorsal pallium derivative. After tracer injections into the dorsal cortex of lizards, we found long descending projections that reached the striatum, the diencephalic basal plate, and the mesencephalic tegmentum, which suggests that it may represent a sensorimotor cortex.     

Acoustically evoked immediate early gene expression in the pallium of female túngara frogs

In anurans, much is known about the role of the auditory midbrain in processing conspecific calls, but comparatively little is known about the role of the pallium. To address this deficiency, we investigated the induction of the immediate early gene egr-1 by natural mate chorus in the medial, dorsal, lateral, and ventral pallium of female túngara frogs. We found strong acoustically evoked egr-1 expression in the dorsal medial pallium (p < 0.01) and ventral pallium (p = 0.02), with a weaker effect in the lateral pallium (p = 0.05). In the ventral pallium, acoustically induced egr-1 expression was stronger in the anterior portion. Measures of movement and olfactory activity could not explain a significant portion of acoustically evoked pallial egr-1 expression. In contrast, egr-1 expression in the auditory midbrain covaried with egr-1 expression in the dorsal medial pallium and ventral pallium, suggesting that their activity was coupled with auditory activity. Taken together, these results suggest that the acoustically evoked egr-1 expression in the dorsal medial pallium and ventral pallium were a direct result of auditory stimulation. Furthermore, although both anatomical and electrophysiological evidence demonstrate that multiple modalities overlap in the frog pallium, our results show that a multimodal stimulus is not required to activate pallial neurons. Although the functional role of the frog pallium is not known, our results demonstrate that species-specific sounds activate spatially segregated and anatomically distinct areas of the frog pallium, inviting further investigation into the role of the frog pallium in acoustic communication.     

Experimental study of the projections of the nucleus of the tractus solitarius and the area postrema in the cat

Projections from the area postrema and adjacent parts of the medial solitary nucleus are demonstrated with the Nauta method following lesions limited exclusively to these structures. Experiments are controlled with lesions involving adjacent bulbar regions, cerebellum, and spinal cord. Ascending pathways in the dorsal and lateral columns of the spinal cord project ipsilaterally to the area postrema and bilaterally to a para-alar nucleus in the ventral periphery of the nucleus gracilis. Neurons in the area postrema project mainly inspilaterally to the dorsal and medial regions of the medial solitary nucleus. Neurons in the posterior half of the medical solitary nucleus project ipsilaterally to the lateral solitary nucleus, dorsal vagal nucleus, ambigus, retrofacial nucleus, and dorsal and lateral bulbar reticular formation. Projections to nuclei intercalatus and prepositus hypoglossi, bilaterally, and to the ipsilateral dorsal tegmental nucleus by way of the dorsal longitudinal fasciculus are also shown. No direct projections to the diencephalon are demonstrated. Control lesions in the dorsal column nuclei reveal projections to the contralateral inferior olive and thalamic reticular and ventrobasal nuclei, but not to the projection sites of the solitary nucleus. Evidence is given to support the hypothesis that ascening visceral pathways are interruped in the bulbar reticular formation and dorsal tegmental nucleus before reaching the diencephalon. Correlations are suggested with functional aspects of the central autonomic and reticular activating systems.     

Understanding the basic circuitry of the cerebral hemispheres: the case of lizards and its implications in the evolution of the telencephalon

The organization of the cerebral hemispheres of mammals is characterized by corticostriatal glutamatergic projections and striatopallidal GABAergic ones, plus the descending projections of the pallium and subpallium to extratelencephalic targets. The present review of the available neuroanatomical data on the forebrain of lizards suggests that the telencephalon of reptiles also follows this basic pattern of connectivity. In addition, we show that this basic circuitry includes a pallido-cortical projection, therefore forming a cortico-striato-pallido-cortical circuit. The analysis of this circuitry for the medial, dorsal, lateral, and ventral pallial divisions in reptiles and mammals leads to the following conclusions: (1) The medial and dorsal cortices of lizards together appear to be equivalent to the medial pallium of mammals. (2) The projection from the lacertilian dorsal cortex to the striatum proper resembles the subiculo-striatal projection of mammals, rather than the isocortical projection to the caudatus-putamen. (3) Most of the dorsal striatum of reptiles is engaged in the corticostriatal circuit corresponding to the ventral pallium (the anterior dorsal ventricular ridge), and therefore, it is not equivalent to the mammalian caudatus-putamen, which is involved in the circuit of the dorsal pallium. (4) The main and accessory olfactory bulbs also follow this pattern of connections.     

Olfactory bulb projections in the bichir, Polypterus

The central projections of the olfactory bulb were studied in Polypterus using the Nauta and Fink-Heimer techniques. Two major target areas were identified in the subpallium: the lateral subpallial nucleus and the dorsal and ventral entopeduncular nuclei. The connections are predominantly, if not exclusively, ipsilateral. In the pallium a massive ipsilateral projection to the superficial third of the medial pallium was demonstrated while the remainder of the pallium was found to be free of degeneration. Thus it appears that the pallium of Polypterus is not uniform throughout, as has been suggested in the literature. This contention is also supported by an analysis of the pallial cytoarchitecture. Because the pallium of Polypterus is everted, rather than inverted and evaginated, the topographically medial pallium is in topological correspondence with the lateral pallium of tetrapods. On the basis of this topological correspondence and the similarity of afference from the olfactory bulb, it is argued that the “medial” pallium of Polypterus is homologous to the pyriform pallium of tetrapods. The findings of this study are compared to those of similar studies in teleosts, which also have an everted pallium. An apparent conflict appears and suggestions for resolving it are offered.     

Connections of the lateral and medial divisions of the goldfish telencephalic pallium

Biotinylated dextran amine and fluorescent carbocyanine dye (DiI) were used to examine connections of the lateral (Dl) and medial (Dm) divisions of the goldfish pallium. Besides numerous intrinsic telencephalic connections to Dl and Dm, major ascending projections to these pallial divisions arise in the preglomerular complex of the posterior tuberculum, rather than in the dorsal thalamus. The rostral subnucleus of the lateral preglomerular nucleus receives auditory input via the medial pretoral nucleus, lateral line input via the ventrolateral toral nucleus, and visual input via the optic tectum, and it projects to both Dl and Dm. The anterior preglomerular nucleus and caudal subnucleus of the lateral preglomerular nucleus receive auditory input via the central toral nucleus and project to Dm. This pallial division also receives chemosensory information via the medial preglomerular nucleus. The central posterior (CP) nucleus, which receives both auditory and visual inputs, also projects to Dm and is the only dorsal thalamic nucleus projecting to the pallium. Thus, both Dl and Dm clearly receive multisensory inputs. Major projections of CP and projections of all other dorsal thalamic nuclei are to the subpallium, however. Descending projections of Dl are primarily to the preoptic area and the caudal hypothalamus, whereas descending projections of Dm are more extensive and particularly heavy to the anterior tuber and nucleus diffusus of the hypothalamus. The topography and connections of Dl are remarkably similar to those of the hippocampus of tetrapods, whereas the topography and connections of Dm are similar to those of the amygdala.     

the connections of the mouse olfactory bulb: A study using orthograde and retrograde transport of wheat germ agglutinin conjugated to horseradish peroxidase

The efferent and centrifugal afferent connections of the main olfactory bulb (MOB) of the mouse were studied by orthograde and retrograde transport of wheat germ agglutinin conjugated to horseradish peroxidase (WGA-HRP). MOB projects ipsilaterally to the anterior olfactory nucleus, taenia tecta, anterior hippocampal continuation, indusium grisium, olfactory tubercle, and the lateral and medial divisions of the entorhinal area. In the region of the anterior one-half to two-thirds of the posterior division of the insular cortex the projection from MOB extends into the insular cortex. The only efferent projection of MOB to the contralateral half of the brain was to the anterior olfactory nucleus. All efferent projections of MOB, thus, are to telencephalic structures. By contrast the centrifugal afferents to MOB originate from every major division of the neuraxis. Neurons projecting to the bulb were found ipsilaterally in all divisions of the anterior olfactory nucleus (AON). In some cases, labeling in the external division of AON was weak or absent. In the contralateral AON, pars externa was the most intensively labeled sub-division. Retrogradely labeled neurons were also present in all other subdivisions of the contralateral AON but were fewer in number and less heavily labeled than in the ipsilateral AON. Ipsilaterally, positive neurons were also present in taenia tecta, and the anterior hippocampal continuation. There was profuse retrograde labeling of neurons in the entire extent of the ipsilateral piriform cortex (PC). There was a rostral to caudal gradient of labeling in PC with more positive neurons in rostral than caudal parts. Labeled neurons were present in the lateral entorhinal cortex LEC and in the transitional cortex between LEC and PC. Very heavy retrograde labeling was present in the nuclei of the horizontal and vertical limbs of the diagonal band (HDB and VDB). More cells were labeled in HDB than in VDB. Neurons were labeled in the ipsilateral nucleus of the lateral olfactory tract (NLOT) and, when the injection spread into the accessory olfactory bulb, labeled neurons were present ventral to NLOT in accessory NLOT. A few lightly labeled neurons were always present in the posterolateral and medial cortical amygdaloid areas. Neurons were labeled in the zona inserta and scattered throughout several hypothalamic nuclei. There was massive retrograde labeling of neurons in the locus coeruleus and neurons were abundantly labeled in the dorsal and medial raphe nuclei and nucleus raphe pontis. In general, the labeling of MOB connections was more extensive than that which has been reported in closely related species.(ABSTRACT TRUNCATED AT 400 WORDS)     

Olfactory projections in the white sturgeon, Acipenser transmontanus: an experimental study

Telencephalic evolution in ray-finned fishes shows increasing complexity from polypteriform fishes through sturgeons to teleosts. Telencephalic organization in sturgeons is thus critical to our understanding of ray-finned fish evolution, but it is poorly understood, particularly as regards the roof or pallium. Two major hypotheses exist regarding the medial part of area dorsalis (Dm): that Dm is extended; and that Dm is restricted. The extent and topography of secondary olfactory projections to the pallium are critical in evaluating these hypotheses, but there is little agreement regarding these projections. Olfactory projections in the white sturgeon were therefore examined by using the carbocyanine probe DiI, biocytin, and biotinylated dextrin amine (BDA). Both DiI and BDA revealed primary olfactory projections to the olfactory bulb and primary extrabulbar projections widely in the telencephalon and to more restricted regions of the diencephalon. Myelinated secondary olfactory fibers caused DiI to be less effective in labeling secondary olfactory projections, which terminate in all subpallial nuclei and in the pallium: sparsely in the medial pallial division (Dm); heavily in the posterior pallial division (Dp); and more lightly in the lateral pallial division (Dl). In the diencephalon, substantial secondary olfactory projections were seen to the habenular nuclei, the rostral pole of the inferior lobe, and several nuclei of the posterior tubercle. All secondary olfactory projections were bilateral but heavier ipsilaterally. Bulbopetal neurons were located in both pallial and subpallial centers and were more numerous ipsilaterally. These results corroborate an earlier experimental study on the shovelnose sturgeon and indicate a restricted Dm in sturgeons.     

Connections of the terminal nerve and the olfactory system in two galeomorph sharks: an experimental study using a carbocyanine dye

In elasmobranchs the terminal nerve courses separately from the olfactory nerve. This characteristic makes elasmobranchs excellent models to study the anatomy and function of these two systems. Here we study the neural connections of the terminal nerve and olfactory system in two sharks by experimental tracing methods using carbocyanine dyes. The main projections from the terminal nerve system (consisting of three ganglia in Scyliorhinus canicula) course ipsilaterally to the medial septal nucleus and bilaterally to the ventromedial telencephalic pallial region. Minor terminal nerve projections were also traced ipsilaterally to diencephalic and mesencephalic levels. With regard to the olfactory connections, our results show that in sharks, unlike ray-finned fishes, the primary olfactory projections are mainly restricted to the olfactory bulb. We also performed tracer application to the olfactory bulb in order to analyze the possible central neuroanatomical relationship between the projections of the terminal nerve and the olfactory bulb. In these experiments labeled neurons and fibers were observed from telencephalic to caudal mesencephalic regions. However, we observe almost no overlap between the two systems at central levels. The afferent and the putatively efferent connections of the dogfish olfactory bulb are compared with those previously reported in other elasmobranchs. The significance of the extratelencephalic secondary olfactory projections is also discussed in a comparative context.     

Olfactory projections in the lepidosirenid lungfishes

Olfactory nerve and olfactory bulb projections in lepidosirenid lungfishes were experimentally determined with neural tracers. Unilateral injections of DiI into the olfactory nerve labeled the accessory and main olfactory bulbs as well as fibers of the anterior root of the terminal nerve, which terminates extensively in cell groups of the medial hemispheric wall, the dorsal and lateral pallia, and the preoptic nuclei and posterior tubercle. Lepidosirenid lungfishes do not exhibit separate vomeronasal nerves, but previous data indicate that calbindin-positive receptors within basal crypts of the olfactory epithelium are homologous to the vomeronasal organ of tetrapods. Unilateral injections of DiI into the accessory olfactory bulb reveal an accessory olfactory tract which terminates primarily if not solely in the ipsilateral medial amygdalar nucleus as in amphibians. Unilateral injections of tracers into the main olfactory bulb reveal extensive projections to all cell groups in the ipsilateral telencephalic hemisphere, except for the medial amygdalar nucleus, as well as secondary olfactory projections (decussating in the habenular commissure) to the contralateral dorsal pallium and main olfactory bulb. Secondary olfactory projections also terminate bilaterally in diencephalic and midbrain centers after partial decussation in the anterior and postoptic commissures, as well as in the ventral hypothalamus and posterior tubercle. Cladistic analysis of the extensive secondary olfactory projections indicates that this pattern is primitive for all bony fishes whereas the reduction in secondary olfactory projections in amphibians, particularly anurans, is a derived, simplified pattern.     

Dorsomedial telencephalon of lungfishes: A pallial or subpallial structure? criteria based on histology,connectivity, and histochemistry

The dorsomedial telencephalon of lepidosirenid lungfishes has been interpreted in two divergent ways: earlier investigators regarded it as a subpallial (septal) structure; more recently, it has been reinterpreted as the medial pallium (hippocampus). To resolve this question, we identified parameters that are conclusive in their association with either the medial pallium or the septum in anamniotes. The present study examines the position of ependymal thickenings and the distribution of acetylcholinesterase (AchE) in the cerebral hemispheres of the African lungfish Protopterus, the Australian lungfish Neoceratodus, and the amphibian species Xenopus and Ambystoma. In addition, projections from the hypothalamus (paraventricular organ) to the telencephalon are investigated in Protopterus. Ependymal specializations are located dorsally and ventrally in the lateral ventricles of amphibians, but laterally and medially in lungfishes. In Protopterus, the paraventricular organ projects to the medial telencephalic hemisphere, but not to the dorsal roof. High levels of AchE are present in restricted neuropil regions of the medial hemisphere and in the ventral and ventrolateral telencephalon, but they are lacking in the dorsal roof. Intensely AchE-stained neuronal cell bodies are located in the ventral telencephalon (rostrally) and the dorsomedial telencephalon (at mid-level). In Neoceratodus, AchE staining is pronounced in the septal area, but absent in the pallium. The terminal nerve proper lacks AchE stain in Protopterus; nerve fibres of the preoptic nerve are AchE-positive in both lungfish species. In Xenopus, AchE staining of fibers and terminals is restricted to the subpallium (medial septum, tuberculum olfactorium, striatum, nucleus accumbens, and medial amygdala); cell bodies are AchE positive in parts of the subpallium and rostral pallium. Comparison of cytological, histochemical, and "connectional" parameters substantiates the interpretation that the dorsomedial telencephalon of lungfishes represents a subpallial, but not a "medial pallial" structure. The dorsomedial part of the lepidosirenid telencephalon corresponds to the septum in the most plesiomorphic living lungfish, Neoceratodus forsteri, but it differs considerably from the dorsomedial telencephalon (medial pallium) in amphibians.     

Neuropeptides and catecholamines in efferent projections of the nuclei of the solitary tract in the rat

This study focuses on the involvement of catecholamines and nine different peptides in efferents of the nucleus of the solitary tract to the central nucleus of the amygdala, the bed nucleus of the stria terminalis, and different parabrachial and hypothalamic nuclei in the rat. A double-labeling technique was used that combines a protein-gold complex as the retrograde tracer with immunohistochemistry. Catecholaminergic projection neurons were the most numerous type observed and projected mainly ipsilaterally to all targets studied. Most projections arose from areas overlying the dorsal motor nucleus, mainly the medial nucleus. Neurons synthesizing somatostatin, met-enkephalin-Arg-Gly-Leu, dynorphin B, neuropeptide Y, and neurotensin projected to all structures examined. Somatostatin and enkephalin immunoreactive projection cells were the most numerous. They were located in close proximity to each other, including all subnuclei immediately surrounding the solitary tract, bilaterally. Most dynorphin and neuropeptide Y immunoreactive projection cells were found rostral to that of enkephalinergic and somatostatinergic projections, and mainly in the ipsilateral medial nucleus. Neurotensinergic projections were sparse and from dorsal and dorsolateral nuclei. Substance P and cholecystokinin contribute to parabrachial afferents. The location of substance P immunoreactive projection cells closely resembled that of enkephalinergic and somatostatinergic projections. Projecting cholecystokinin immunoreactive cells were observed in dorsolateral nucleus. Bombesin immunoreactive cells in dorsal nucleus projected to either the parabrachial or hypothalamic nuclei. No vasoactive intestinal polypeptide-containing cells were detected. Thus, most catecholaminergic and neuropeptidergic efferents originated from different populations of cells. It is proposed that catecholaminergic neurons constitute the bulk of solitary efferents and that they may contribute to autonomic neurotransmission. Peptidergic neurons mainly form other subgroups of projections and may play a role in modulating the physiological state of the target nuclei.