Ventromedial mesencephalic tegmental (VMT) projections to ten functionally different cortical areas in the cat: topography and quantitative analysis.

The cortical projections of the ventromedial mesencephalic tegmentum (VMT) were studied in the cat by the retrograde transport of wheat germ agglutinin-conjugated HRP (WGA-HRP). The following cortical regions were injected: prefrontal, cingulate, motor, somatic sensory, auditory, primary visual, secondary visual, parasplenial, insular, and entorhinal. Labelled cells in the five component nuclei of the VMT (rostral linear, parabrachial pigmented, central linear, interfascicular, and paranigral nuclei) were counted and the numbers correlated with the total cell populations of the nuclei. In all experiments large numbers of cells were labelled in the VMT, particularly in the rostral linear and parabrachial pigmented nuclei. Following entorhinal cortical injections, significant labelling occurred in the interfascicular and central linear nucleus, although some cells of these nuclei were labelled in every experiment. The cells of the paranigral nucleus remained unlabelled in all experiments. Several hundred VMT cells were labelled in most experiments, over 1000 in the case of the parasplenial and over 2000 in the case of prefrontal injections. The most strongly labelled nucleus was the rostral linear, in which up to 36% of the total cell population could be labelled in a single experiment. These results indicate that the VMT of the cat is a major source of brainstem projections to the cerebral cortex and that all cortical areas are reached by this system. Although the projections to the prefrontal and limbic cortical areas are particularly prominent, there appears to be a consistent and significant projection to all areas. Further, the projection of the VMT to the cortex is topographically organized, indicating that the functional roles of the component nuclei of the VMT may be substantially different.

Early development of serotonin-containing neurons and pathways as seen in wholemount preparations of the fetal rat brain.

The early development of serotonin-containing neurons was studied in wholemounts of the fetal rat brain (E12-E18). The wholemounts were treated immunocytochemically according to an immunoperoxidase technique to reveal a panorama of developing serotoninergic neurons. Serotoninergic neurons were localized to two discrete groups or clusters within the brainstem. Serotonin-containing neurons were identified first at E12 forming a rostral cluster of cells just caudal to the mesencephalic flexure. The more caudal cluster of cells first appeared at E14 in the medulla. During the period from E12 to E18, the immunoreactive cells increased in number and acquired a more complex dendritic tree while migrating to their permanent position. At E16, cells of the rostral group exhibited remarkably uniform mediolateral orientation. The rostral group of immunoreactive neurons gave rise to almost all ascending fibers, whereas the caudal group gave rise to the majority of descending fibers. Growing serotoninergic fibers were tipped by prominent growth cones, which were strongly immunoreactive. The fibers demonstrated prominent orientational selectivity with an almost total separation into ascending and descending bundles. Some of the ascending immunoreactive fibers displayed acute changes in their direction of growth, suggesting that the orientation of serotininergic fibers is mediated by directional cues that are specific to particular populations of serotoninergic fibers. Serotoninergic axons within the medial forebrain bundle were demonstrated particularly well and their ascent and rate of growth toward the forebrain could be easily followed. Immunoreactive fibers entered the telencephalon at E17 two portals, one along the lateral border of the hypothalamus and one rostrally, adjacent to the olfactory tubercle. In wholemounts at E18, fibers arising from this latter location could be followed as two distinct bands within the pallium; a basal band located ventrolaterally, adjacent to the lateral olfactory tract, and a dorsal band located at the medial edge of the telencephalon. Both fascicles were directed toward the occipital pole and contained unbranched fibers. At E18, serotoninergic axons arising from these two loosely organized fascicles covered most of the frontal telencephalon. The results of the present study indicate that wholemounts of embryonic brain can provide novel spaciotemporal data on the development of neuro-transmitter systems and may in the future prove to be useful experimental preparations in developmental neurobiology.

Comparative anatomy of the ventromedial mesencephalic tegmentum in the rat, cat, monkey and human.

The five component nuclei of the ventromedial mesencephalic tegmentum (VMT) were studied on Nissl stained serial sections of the brain stem of rat, cat, monkey (Macaca nemestrina) and human. Models of the VMT nuclei were constructed to compare their size, shape and disposition across species. For each nucleus in each species the following were calculated: the volume, the number of neurons, the size distribution of neurons, the mean soma size and the packing density of neurons. The morphology of the cells in the different nuclei is also described. The parabrachial pigmented nucleus (PBP) forms, on average, 51% of the VMT volume and cell number. The paranigral nucleus (PN) and the central linear nucleus (LC) formed 19% and 14% of the VMT volume and cell number respectively. The relatively small, but compact interfascicular nucleus (IF) was on average 9% of the VMT volume and cell number and the rostral linear nucleus (LR) formed its remaining 7%. However, in different species the relative prominence varies between species. Thus PBP is the largest of the VMT nuclei in the monkey, PN is particularly well developed in the human, IF contains a particularly large number of cells in the rat, and LR and LC are strongly developed in the cat. This study presents a cytoarchitectonic description of the five nuclei in each species. The distinctive cytoarchitectonic appearance of each nucleus suggests that their functions may differ. This possibility, which is strengthened by evidence that the projections of the VMT nuclei are differential, may need to be considered in the interpretation of the results of experimental investigations using stimulation and/or lesion experiments in the VMT region and in the interpretation of pathological findings in the human brain.

Serotoninergic innervation of the cat cerebral cortex.

Serotoninergic axons in the cat cerebral cortex were demonstrated immunohistochemically with a monoclonal antibody to serotonin (5-HT). Three types of 5-HT axons are distinguished at the light microscopic level by differences in their morphology. Small varicose axons are fine (less than 0.5 micron) and bear fusiform varicosities that are generally less than 1 micron in diameter. These axons extend throughout the width of the cortex and branch frequently, giving rise to widely spreading collaterals. Nonvaricose axons are smooth, show a relatively large and constant caliber (about 1 micron), travel in straight, horizontal trajectories, and branch infrequently. Large varicose axons are distinguished by large round or oval varicosities (1 micron or more in diameter) borne on fine-caliber fibers. These axons often form basket-like arbors around the somata of single neurons. In the simplest basket-like arbors, several large, round varicosities from a small number of axons contact the soma. In complex baskets intertwining collaterals contact the soma and apparently climb along and outline the cell's major dendrites. The patterns revealed by the climbing axons suggest that a variety of nonpyramidal cell types selectively receive dense 5-HT innervation. Serial reconstructions of the 5-HT axons within the cortex show that the large varicose axons arise as infrequent collaterals from the nonvaricose axons. A single nonvaricose parent axon gives rise to several large varicose axon collaterals that may contribute to different basket-like arbors. Conversely, a single basket-like arbor may be formed by large varicose axon collaterals from more than one nonvaricose parent axon. The small varicose axons do not appear to be related within the cortex to either the nonvaricose or large varicose axon types. The results support the hypothesis that the 5-HT projection to the cortex is organized into two subsystems, one of which may exert widespread influence in the cortex via highly divergent branches, while the other, with a more restricted distribution, acts on specific classes of cortical neurons.

Electron microscopic analysis of the mesencephalic ventromedial tegmentum in the cat.

We have studied the normal ultrastructure of the ventral mesencephalic tegmentum (VMT) in the cat, particularly the morphology and distribution of presynaptic terminals and the types of synaptic junctions. The following subnuclei of the region were examined: n. linearis rostralis (LR), n. paranigralis (PN), and n. interfascicularis (IF). The qualitative and quantitative data revealed significant ultrastructural differences between these subnuclei. Each subnucleus had a characteristic dendritic structure. In LR the dendrites were nonspinous and cylindrical and had presynaptic terminals randomly distributed over their surface. In PN we observed varicose dendrites with spines; the presynaptic terminals formed clusters on the narrow segments of the dendrites and around the spines. Dendrodendritic synapses were also observed in this nucleus. In IF, there was an internal division regarding dendritic structure: in the rostral part of the nucleus there were cylindrical dendrites while in the caudal part irregularly shaped dendrites bearing long spines were found. In IF and LR some of the cylindrical dendrites were seen to be in direct contact with the basal lamina of blood vessels. Four types of presynaptic terminals were distinguished by the morphology of their vesicles, and the proportion of each type in the total terminal population was determined. On this basis the compositions of the presynaptic terminal population in the three subnuclei were found to be very similar. Most terminals contained clear, round vesicles (62.6%), or both clear and dense-cored vesicles (35.1%). Few terminals were seen with dense-cored vesicles only (1.4%) or with pleomorphic vesicles (0.9%). The majority of synapses in the VMT were found to have symmetrical densities. LR had twice as many asymmetrical synapses as the other two subnuclei. Eighty percent of the terminals formed synapses with dendrites, although axosomatic and axoaxonic synapses were also seen. The density of the terminals was significantly different for each subnucleus: 191/1,000 micrometers 2 in IF, 120/1,000 micrometers 2 in PN, and 81/1,000 micrometers 2 in LR. These data indicate that while the subnuclei of the VMT receive morphologically similar afferents, each has a unique way of processing the information provided by them, through a different internal circuitry.

Differential development of the dual serotoninergic fiber system in the cerebral cortex of the cat.

The changes in distribution and density of the serotoninergic innervation of the cerebral cortex were studied in kittens from birth (PO) to 60 days of age (P59). Three cortical areas were sampled: prefrontal, primary auditory, and primary visual areas. Two systems of serotoninergic axons were demonstrated by immunocytochemical techniques: the fine axon system characterized by small fusiform varicosities up to 1 micron in diameter, and the beaded axon system, the fibers of which have round varicosities up to 5 microns in diameter. The density of the two types of fibers across the cortical layers at different ages was measured with a semiautomatic computerized system. In all three areas, the density of fine axons increased steadily from birth, although the pattern of innervation changed from an even distribution at PO to a distinct concentration of the fibers in layers I-III by week 2 in the prefrontal cortex and by week 3 in auditory and visual cortices. By contrast, the beaded axons first appeared in the cortex at week 2 for the prefrontal cortex, at week 3 in auditory and visual areas. Initially, these fibers were distributed throughout all cortical layers and were of much lower density than the fine axons. At later ages the beaded axons became confined to layers I-III where they gradually increased in number, and from week 4, they formed pericellular arrays which were only observed in the prefrontal and auditory cortices, not in visual cortex. These findings provide further evidence for the existence of two parallel subsystems of serotoninergic axons which are different not only in their morphology and nuclear origin, but also in their development. Our finding that the two serotoninergic fiber systems arrive in the cortex in two different stages suggests that they have differential roles in development. The late formation of the pericellular arrays indicates that the formation of the specific connections made by the beaded fibers could be dependent on a certain degree of maturity of the target neurons.

Distribution of substance P-like immunoreactive neurons and terminals throughout the nucleus of the solitary tract in the human brainstem.

The anatomical distribution of substance P-like immunoreactivity across the subnuclear divisions of the nucleus of the solitary tract has been examined in the human medulla oblongata. A differential distribution of neurons, fibres, and terminals was observed throughout the ten subnuclear divisions of this nucleus. Substance P-like immunoreactive neurons were observed most frequently in the nucleus gelatinosus, with moderate numbers in the medial, intermediate subnuclei and very few in the commissural, ventral, dorsal, and dorsolateral subnuclei. The paracommissural, ventrolateral, and interstitial subnuclei did not contain substance P-like-immunoreactive neurons. These neurons were typically bipolar and moderate-sized to large, except for the neurons in the nucleus gelatinosus, which were substantially smaller. The highest densities of fibres and terminals were observed in the gelatinosus, medial, and intermediate nuclei, with moderate densities in the paracommissural and dorsal subnuclei. Sparse substance P-like-immunoreactive fibres and terminals were seen in the ventral and interstitial nuclei as well as within the solitary tract. The dorsolateral nucleus was characterized by a light distribution of fibres and terminals, except for a dense aggregation along its lateralmost border. A prominent innervation of pigmented neurons by substance P-like-immunoreactive terminals and fibres was also observed in the dorsolateral nucleus. The results reveal that the subnuclear complexity of the nucleus of the solitary tract is richly reflected by its differential pattern of substance P-like-immunoreactive structures.

The afferent ganglion cells and cortical projections of the retinal recipient zone (RRZ) of the cat's pulvinar complex'.

A retino-pulvinar projection in the cat was confirmed using anterograde (autoradiography) and retrograde (horseradish peroxidase (HRP)) tracing techniques. The part of the "pulvinar complex" receiving retinal afferents is referred to as the retinal recipient zone (RRZ). The cortical projections of the RRZ were studied by injecting HRP into different cortical areas. The retrograde labeling of the cell bodies and dendritic fields of the retinal ganglion cells projecting to the RRZ was accomplished by injecting HRP electrophoretically into the RRZ. Our results indicate that the RRZ projects to areas 19 and the lateral suprasylvian area (LS) but not to areas 17 or 18. Virtually all RRZ cells, including those that project to the cortex, are small (10-20 micron in diameter); they are the same size as the relay cells of the parvocellular C laminae of the lateral geniculate nuclear complex (LGNd) that project to areas 19 and LS. The majority of the ganglion cells projecting to the RRZ had medium-sized somas (15-25 micron in diameter), large (up to 800 micron in diameter), diffuse dendritic fields with a characteristic morphology, and appeared different from the alpha, beta, gama, and delta cells of Boycott and Wässle ('74). These cells provide evidence for another morphological class of ganglion cells, termed epsilon cells. Our results suggest that the RRZ relays the activity of specific types of retinal ganglion cells to extrastriate visual cortex and, thus, functions in parallel with the different subdivisions of the LGNd.

Dorsal motor nucleus of the vagus nerve: a cyto- and chemoarchitectonic study in the human.

In order to investigate the topography and subdivisions of the human dorsal motor nucleus of the vagus nerve (10), studies were conducted using cyto- and chemoarchitectonic (acetylcholinesterase and substance P-like immunoreactivity) and computer reconstruction techniques. The six brainstems examined were obtained within 17 hours postmortem from adults with no known neurological disorders. Serial sections cut in transverse, sagittal, and coronal planes were stained with cresyl violet, or tested for acetylcholinesterase or substance P. The neurons of the 10 (16,826 +/- 967) displayed cyto- and chemoarchitectonic heterogeneity and could be classified into six types. Types I-V consist of presumed vagal motor neurons (13,802 +/- 844), while the remaining type (Type VI) consisted of presumed interneurons (3,024 +/- 769). The 10 was subdivided into nine subnuclei grouped regionally into rostral, intermediate, and caudal divisions on the basis of neuronal morphology, cell density, and differential AChE and substance P reactivities. The rostral division contains the dorsorostral (DoR) and the ventrorostral (VeR) subnuclei; the intermediate division contains the rostrointermediate (RoI), dorsointermediate (DoI), centrointermediate (CeI), ventrointermediate (VeI), and caudointermediate (CaI) subnuclei; the caudal division (Ca) is not subdivided. Morphologically, small round or oval cells populate the VeR and VeI. Medium-sized oval cells occur in the DoR, CeI, and Ca, while medium-sized fusiform and multipolar cells are the main features of CaI. Large triangular cells appear mainly in DoI. Glial cells show the highest predilection for CeI, lowest densities in DoI and medial fringe subnucleus (MeF), and intermediate densities in the remaining six subnuclei. VeI showed the strongest AChE reactivity. Although the cell bodies of VeR and DoI are AChE positive, the neuropil (background) is weakly stained. Densely distributed fine granular substance P-like immunoreactivity occurs throughout the entire nucleus, while the intermediate and caudal divisions contain substance P-like-immunoreactive neurons. Three-dimensional computer reconstructions afforded an appreciation of the distinctiveness of the intermediate division (a division that contains the majority of cells) and the caudal division, which displays the lowest density of presumed vagal motoneurons. It is possible that the subnuclei identified herein form functional units innervating specific organs.

Cytoarchitecture of the human dorsal raphe nucleus.

Serial 50 microns Nissl-stained sections through the midbrain and pontine central gray of four adult humans (mean age 56 years, mean postmortem delay 3 hours) were analysed and the subnuclei of the dorsal raphe nucleus (DR) delineated on the basis of neuronal morphology and density. Five subnuclei were apparent: the interfascicular, ventral, ventrolateral, dorsal, and caudal. The area of each subnucleus was measured in sections selected at regular intervals throughout the length of the DR. The number of neurons was counted and their density within each subnucleus calculated. The dorsal subnucleus was the largest and contained the majority of neurons but had the lowest neuronal density. The ventrolateral subnucleus had the highest density of neurons. A total of 235,000 +/- 15,000 neurons (average of 1,200 +/- 200 neurons per section) were found within a volume of 71.3 +/- 4.5 mm3 of DR with a mean neuronal density of 3,300 +/- 200 neurons/mm3. Morphometric and morphological analysis of DR neurons revealed four distinct neuron types: round, ovoid, fusiform, and triangular. These types of neurons characterized particular subnuclei. The location and boundaries of the subnuclei of the human dorsal raphe are presented in the form of an atlas. The subdivisions described are similar to that described in other mammals. On the basis of this information the location of particular projection neurons within the human dorsal raphe can be predicted and the effects of disease on this nucleus may be forecast.

Distribution of two morphologically distinct subsets of serotoninergic axons in the cerebral cortex of the marmoset.

The serotoninergic innervation of the marmoset (New World monkey, Callithrix jacchus) cerebral cortex has been analyzed by using immunocytochemistry. The use of a sensitive monoclonal antibody against serotonin allowed the visualization of the fine morphology of individual axons. Two types of terminal axons were demonstrated: one has sparse, small, ovoid varicosities (dia. less than 1 micron), and the other has large, spheroidal varicosities (up to 5 microns in dia.), which are more densely clustered. The first type of axon is distributed through all cortical layers, with a characteristic laminar distribution that varies from area to area. The second type of axons was distributed sparsely in all regions but was markedly denser in the frontal and anterior parietal lobes, and in the hippocampal formation. Axons with large varicosities typically surrounded certain cell bodies and proximal dendrites, forming pericellular arrays, or baskets. These morphological specializations were most frequent in the frontal and anterior parietal cortex, where they were found around stellate and horizontal cells in layer I and around stellate and bipolar cells in layer II and III. Similar baskets were also found in the hippocampal formation, mainly along the border between the hilus and the granule cell layer of the dentate gyrus, across the CA4 field, and at each side of the pyramidal cell layer of the CA3 regions. The distribution and cellular morphology of the cell surrounded by the 5-HT basket fibres were suggestive of a subpopulation of interneurons, possibly GABAergic and/or peptidergic. In agreement with previous reports on the innervation of the cerebral cortex of other mammalian species, the marmoset cerebral cortex is innervated by two separate subsystems of serotoninergic axons. One of these may have a strong and specific influence on the cortical inhibitory circuitry, via relay through cortical interneurons.

Substance P- and tyrosine hydroxylase-containing neurons in the human dorsal motor nucleus of the vagus nerve.

The aim of this study was to provide a comprehensive account of the topography, morphology, and frequencies of the substance P- and tyrosine hydroxylase-containing neurons in the human dorsal motor nucleus of the vagus nerve. The morphology of immunoreactive neurons was studied and the variations of the cell distributions were presented by three-dimensional computer reconstructions. Three types of substance P-like immunoreactive neurons were identified. They were predominantly located in the dorsointermediate, centrointermediate, caudointermediate, and caudal division of the dorsal motor nucleus of the vagus nerve. The morphology of substance P-like immunoreactive neurons varied according to the subnuclei in which they were found. Three types of tyrosine hydroxylase-like immunoreactive neurons were identified, mainly in the periphery of the dorsal motor nucleus of the vagus nerve, including the medial fringe, ventrointermediate, and dorsointermediate subnuclei of the 10. Many cells throughout the ventrointermediate subnucleus of the dorsal motor nucleus of the vagus nerve are seen ventrally to intermingle with the tyrosine hydroxylase neurons of the intermediate reticular zone. Computer reconstructions provided a three-dimensional view of the positions of substance P- and tyrosine hydroxylase-like immunoreactive neurons within the subdivisions of the dorsal motor nucleus of the vagus nerve. The uneven distribution of substance P- and tyrosine hydroxylase-like immunoreactive neurons within the subdivisions suggests an involvement of these substances in some, but not all, autonomic functions of the dorsal motor nucleus of the vagus nerve.

The dorsal, posterodorsal, and ventral tegmental nuclei: a cyto- and chemoarchitectonic study in the human.

In order to verify the existence of the ventral and posterodorsal tegmental nuclei and to extend previous findings regarding the dorsal tegmental nucleus in the human brainstem, studies were conducted using cyto- and chemoarchitectonics, and computer reconstruction techniques. Serial sections of five brainstems from adults with no known neurological disorders were stained for Nissl substance, acetylcholinesterase, and substance P. The topography, cytoarchitecture, and acetylcholinesterase reactivity of the tegmental nuclei were presented in a mini-atlas depicting sections cut in transverse and sagittal planes. The dorsal and posterodorsal tegmental nuclei were identified fully within the central grey matter while the ventral tegmental nucleus extended across the medial longitudinal fasciculus into the pontine reticular formation. The dorsal tegmental nucleus featured a cell-poor pericentral part, strongly positive for acetylcholinesterase, and a central part comprised of densely packed small neurons that displayed moderate acetylcholinesterase reactivity and strong substance P-like immunoreactivity. The posterodorsal tegmental nucleus, located in the same transverse plane as the rostral part of the motor nucleus of the trigeminal nerve, was composed of diffusely arranged small to medium neurons with its neuropil displaying moderate acetylcholinesterase reactivity and strong substance P-like immunoreactivity. The ventral tegmental nucleus, identified as a prominent structure in the pontine tegmentum immediately rostral to the genu of the facial nerve, contained predominantly large neurons and displayed intensive acetylcholinesterase reactivity and substance P-like immunoreactivity. These studies showed that the tegmental nuclei, which displayed distinctive cyto- and chemoarchitectonic features, were fully present in adult human brainstem.

Morphology and distribution of neuropeptide-containing neurons in human cerebral cortex.

Biopsies of human cerebral cortex were fixed by immersion and immunostained for the detection of neuropeptides in neuronal cell bodies and axons. Four neuropeptides (neuropeptide Y, somatostatin, , substance P and cholecystokinin) were visualized in a series of adjacent sections. All populations of immunoreactive neurons had a morphology characteristic of interneurons, with variations in dendritic arborizations and laminar distribution. The cholecystokinin-immunoreactive neurons were most numerous in the supragranular layers, whereas neurons containing the other three peptides occurred mainly in infragranular layers, or even in neurons populating the subcortical white matter. Quantitatively, each population of neuropeptide-containing neurons accounted for 1.4-2.5% of the total neuronal population. The distribution of these neurons varied slightly between cytoarchitectonic divisions, with substance P- and somatostatin-immunoreactive neurons dominating in the temporal lobe and cholecystokinin-immunoreactive neurons in the frontal lobe. Neuropeptide Y-immunoreactive neurons dominated in the gray matter of the frontal half of the hemisphere and in the subcortical white matter of the caudal half of the hemisphere. Furthermore, co-existence of neuropeptide Y or substance P immunoreactivity within somatostatin-immunoreactive neurons could be demonstrated using double labeling immunofluorescence techniques. The axonal plexuses immunoreactive for neuropeptide Y, somatostatin, or substance P were distributed in all layers, with a strong predominance of horizontally oriented fibers in layer I, a moderate plexus of randomly oriented fibers in the supra- and infragranular layers, and a slightly weaker innervation of layer IV. Immunoreactive axons formed, in addition, complex terminal arbors, mostly in older subjects, suggesting that they resulted from an as yet undefined aging process. The present study underlines several aspects of the organization of the neuropeptide-containing neurons of the human cerebral cortex, which are of particular interest in the light of the involvement of these neurons in several neurodegenerative diseases.

Distribution of substance P-like immunoreactive fibres and terminals in the medulla oblongata of the human infant.

This study provides a detailed report of the distribution and density of substance P-like immunoreactive fibres and terminals within the human infant medulla. Seven brains with no signs of macroscopic alteration fixed usually within 24-48 h after death were used. Free floating transverse sections (50-60 microns) were then immunostained with a monoclonal antibody against substance P using the avidin-biotin-peroxidase technique. Morphologically three types of substance P-like immunoreactive fibres were observed: those with small varicosities of less than 1 micron, those with medium varicosities of 1-2 microns and those with large varicosities of 2-4 microns. Very dense substance P-like immunoreactivity was present within the spinal trigeminal nucleus, parts of the gracile and cuneate fasciculi and the paracommissural subnucleus of the solitary nucleus; dense substance P-like immunoreactivity was present within the dorsal nucleus of the vagus nerve, commissural, medial, dorsal, dorsolateral ventral and ventrolateral subnuclei of the solitary nuclear complex, parasolitary nucleus, raphe obscurus, inferior olivary complex (medial, dorsal, dorsomedial nuclei) and ventrolateral part of the dorsal reticular nucleus; moderate substance P-like immunoreactivity was present within the gelatinosus nucleus of the solitary tract, lateral reticular nucleus proper, intermediate reticular zone and parvocellular reticular nucleus; sparse substance P-like immunoreactivity was present within the hypoglossal nucleus, retroambigual nucleus and much of the reticular formation (dorsal, parvocellular, ventral gigantocellular, dorsal paragigantocellular nuclei): and very sparse substance P-like immunoreactivity was present within the nucleus ambiguus, medial vestibular nucleus and parts of the reticular formation (ventral, medial, gigantocellular, ventral gigantocellular, dorsal paragigantocellular nuclei). Substance P-like immunoreactivity was absent in the area postrema, intercalated nucleus, gracile and cuneate nuclei, in the epiolivary, subtrigeminal and parvocellular divisions of the lateral reticular nucleus, spinal vestibular nucleus, and in the solitary and pyramidal tracts. In several regions the substance P-like immunoreactive fibres formed distinct pericellular arrays around the somata and dendrites of neurons (hypoglossal nucleus, dorsal nucleus of the vagus nerve, retroambigual nucleus, intermediate reticular zone). The results indicate the high specificity of the localization of substance P in various structures of the brainstem and underline the presumed significance of this peptide in autonomic and sensorimotor functions of the brain.

Distribution, morphology and number of monoamine-synthesizing and substance P-containing neurons in the human dorsal raphe nucleus.

The distribution, morphology and number of serotonin-, catecholamine- and substance P-containing neurons in the human dorsal raphe nucleus were studied. Parallel series of sections were prepared from 10 human brainstems obtained at autopsy from patients without neurological disease aged between 42 and 88 years. The neurons were identified using immunohistochemistry with antibodies raised against phenylalanine hydroxylase (tryptophan hydroxylase-containing, serotonin neurons), tyrosine hydroxylase (catecholamine neurons) and substance P. A reference series of Nissl-stained sections was also prepared and data published separately were used to delineate the subnuclear divisions of the dorsal raphe nucleus and to establish the total number of neurons in each subnucleus. The following principal findings emerged. (1) Serotonin-synthesizing neurons are present in all regions of the dorsal raphe nucleus and their total number is 165,000 +/- 34,000. The same types of neurons as those seen in Nissl material characterize each of the five subnuclei (caudal, dorsal, ventral, ventrolateral and interfascicular). (2) Substance P-containing neurons mostly occupy the rostral part of the nucleus and their number is 74,600 +/- 17,600. (3) Catecholamine cells are only found in the rostral part of the dorsal raphe nucleus and their number is 5600 +/- 3400. (4) In the ventral and interfascicular subnuclei the combined number of serotonin-synthesizing and substance P-containing neurons exceeds the total number of Nissl-stained neurons suggesting that serotonin and substance P co-exist in a substantial part of the cell population of the dorsal raphe nucleus. This is further supported by the highly similar morphology and size of these neurons. It is concluded that there are demonstrable chemical differences between the various subregions of the human dorsal raphe nucleus. These differences are in harmony with the results of hodological studies in animals, which have demonstrated differential projection pathways emerging from this nucleus.

Morphology of catecholamine-containing amacrine cells in the cat's retina, as seen in retinal whole mounts.

The morphology of catecholaminergic cells of the cat's retina was studied by fluorescence microscopy of retinal whole mounts. Although varied in soma shape, these cells seem to represent a single group of cells with an average soma diameter of 14.5 micrometer in freeze-dried material. The fluorescent terminals of these cells formed a striking two-dimensional pattern: a significant portion of them appeared to be arranged in rings located at the boundary of the inner plexiform and inner nuclear layers. The mean diameter of these rings was 9.7 micrometer and their pattern appeared to extend unbroken across the retina. It was, in general, not possible to observe the connection between these rings and fluorescent somas, except when a ring was in a juxtasomatic position. It is suggested that the postsynaptic somas within these rings are either of other amacrines or of interplexiform cells. Using the Golgi-Colonnier technique on retinal whole mounts, an attempt was made to identify the cell type which resembles the catecholaminergic cell in its size, location and morphology. The suggested cell type appears to be among the largest amacrine cells, (mean soma size of 11.5 micrometer in Golgi material) with a considerable dendritic network at the border of the inner plexiform and inner nuclear layers, although there are branches reaching as far as the middle of the inner plexiform layer.

Distribution of corticospinal motor fibres within the cervical spinal cord with special reference to the phrenic nucleus: a WGA-HRP anterograde transport study in the cat.

The anterograde transport of wheat germ agglutinin conjugated with horseradish peroxidase (WGA-HRP) was used to demonstrate the corticospinal fibres which originate in the motor cortex and project to the cervical spinal cord, specifically to the phrenic nucleus, in the cat. Following injections of WGA-HRP into the pericruciate cortex large numbers of fibres were labelled in the contralateral lateral and ventral funiculi and fewer fibres were labelled in the ipsilateral lateral and ventral funiculi. Labelled corticospinal motor fibres entered the gray matter laterally in laminae V and VI and terminated within these two laminae as well as throughout the entire extent of lamina VII. A few labelled fibres were present in medial lamina VIII and also in lamina IX where they were in close association with the phrenic motoneuron pool. Labelling was present in the gray matter at both sides, with a stronger labelling contralaterally. Labelled axons were also seen crossing from each side of the gray matter to the other side. The results suggest that in the cat the corticospinal motor fibres have a wider distribution in the spinal gray matter than has been previously shown, and that corticospinal motor axons may be in direct contact with phrenic motoneurons.

Histochemical evidence for a catecholaminergic (presumably dopaminergic) projection from the ventral mesencephalic tegmentum to visual cortex in the cat.

An histochemical technique combining the retrograde transport of horseradish peroxidase (HRP) and catecholamine histofluorescence was used to identify those neurons of the cat's mesencephalic ventromedial tegmentum (VMT) which project to visual cortex. In a region corresponding to the ventral tegmental area-A10 nucleus we identified a group of fluorescent cells which were retrogradely labeled by HRP injections into visual cortex. It is suggested that these cells belong to a hitherto unknown, posterior part of the mesocortical dopaminergic system.

Midbrain periaqueductal grey region in the cat has afferent and efferent connections with solitary tract nuclei.

Wheat germ agglutinin-horseradish peroxidase (WGA-HRP) injections were made at the same sites within the midbrain periaqueductal grey region (PAG) of the cat at which microinjections of excitatory amino acids had previously elicited the set of autonomic and somatic reactions (i.e. pupil dilatation, piloerection, retraction of the ears, arching of the back, hissing, howling and growling) known as the 'defence reaction'. The WGA-HRP injections revealed that this PAG region has an extensive set of afferent and efferent connections with solitary tract nuclei (NTS). Within the NTS the majority of labelled neurons were distributed, in approximately equal numbers, in the ipsilateral medial solitary nucleus (SM) and the ipsilateral and the contralateral ventrolateral solitary nuclei (SVL). The densest anterograde labelling was found in the ipsilateral SM, with lighter anterograde labelling in the contralateral SM and bilaterally in the commissural solitary nucleus and SVL. The described connections between the defence region of the PAG and the NTS not only provide a new anatomical basis for cardiovascular and respiratory components of the reaction, but also add to the evidence that, in addition to the hypothalamus, the PAG is an important integrating center for the autonomic and somatic elements of the defence reaction.