Retinal projections in the caecilian Ichthyophis kohtaoensis (Amphibia,Gymnophiona)

Summary The retinal projections of the caecilian Ichthyophis kohtaoensis were investigated by anterograde transport of HRP. The optic tract forms two bundles in the diencephalon, a narrow medial bundle in the optic tectum, and a basal optic tract consisting of few fibres. Terminal fields are in the thalamus, pretectum, tectum, and as a circum-scribed basal optic neuropile in the tegmentum. Thalamic, pretectal and tectal projections are contralateral as well as ipsilateral. The reduced but existing visual projection corresponds to a reduced but existing visually guided behaviour.     

Olfactory and vomeronasal projections and the pathway of the nervus terminalis in ten species of salamanders

Summary Primary olfactory and vomeronasal projections as well as the pathway of the nervus terminalis were studied in 10 representative species of salamandrid and plethodontid salamanders by means of injections of horseradish peroxidase and examination of whole-mount preparations. Olfactory projections are very similar in the different urodeles, but vomeronasal projections differ in shape and number of termination fields. Whereas the direct-developing Plethodontini and Bolitoglossini reveal only one or two fields, the salamandrid species and the members of the plethodontid tribes Desmognathinae and Hemidactyliini, all possessing an aquatic larval stage, exhibit several vomeronasal projection fields. In all species examined centrifugal axons of the nervus terminalis leave the olfactory projection area ventrocaudally and terminate in the preoptic region and the hypothalamus.Abbreviations COM. ANT commissura anterior - DGL displaced glomeruli - HY hypophysis - HYTH hypothalamus - LF lateral fibers of the nervus terminalis - ME medulla oblongata - MF medial fibers of the nervus terminalis - Nt nervus terminalis - Npo nucleus praeopticus     

Central projections of labellar taste hairs in the blowfly,Phormia regina Meigen

Summary We have traced the central projections of the receptor neurons associated with each of the eleven largest taste hairs on the labellum of the blowfly, Phormia regina (Meigen), by staining them with cobaltous lysine. The eleven hairs fall into three groups which reflect their peripheral locations and their branching patterns in the subesophageal ganglion. Group 1, consisting of the anterior hairs (numbers 1 and 2) and Group 3, consisting of the posterior hairs (numbers 9–11) project bilaterally, while Group 2, consisting of the middle hairs (numbers 3–8) projects primarily ipsilaterally. The central projections of the hairs within a single group are similar. Each hair houses four chemoreceptors, which have differing chemical sensitivities and behavioral roles, and one mechanoreceptor. In some cases, there were indications that the different cells within a single hair have different central branching patterns. For some hairs, however, it was clear that a single central branching region and pattern was shared by more than one receptor cell. We failed to find either a continuous somatotopic representation of a hair's position on the periphery, or an anatomical segregation of receptors coding for different modalities. Behavioral experiments indicate that the fly is informed both of the identity of the hair stimulated and of the chemical nature of the stimulus. Our results suggest that this information is not represented on a gross anatomical level.     

The topological organization of primary afferents in the terminal ganglion of crayfish,Procambarus clarkii

Summary The central projections of primary afferents in the terminal ganglion of the crayfish can be seen when an axonal filling with nickel chloride with subsequent silver intensification was used for identification. We describe here the topological relationships of the projections to the landmark structures of the neuropil.The terminal ganglion has five pairs of sensory nerves associated with the mechanosensory hairs and internal proprioceptors. The projection fields of the primary sensory neurons in the nerves Rl and R2 are almost entirely restricted to the ipsilateral half of the ganglion, whereas those of the nerves R3, R4 and R5 cross the midline to form three sensory commissures, A6SCI, A7SCI and A7SCII. The projection fields are segregated from each other, although all are restricted to the ventral neuropil which lies under the ventral intermediate tract (VIT). The intersegmental projections that ascend via the connective ipsilateral to their origins could be observed. This pattern of projection correlates well with the receptive fields exhibited by several mechanosensory interneurons on the body surface of the final segment.     

A critical evaluation of the relationship between the presynaptic network,synaptic vesicles and dense projections in central synapses

Summary Synapses of the oculomotor nucleus of Echidna have been examined ultrastructurally with the aim of integrating data obtained from osmicated and nonosmicated PTA stained material. Particular emphasis has been laid on the relationship between the synaptic vesicles of the osmicated material and the presynaptic network and vesicular grid of the PTA material. This relationship has been explored qualitatively by examining osmicated material of varying qualities of fixation. Such material contains dense projections in addition to synaptic vesicles, and various vesicular network appearances. A variety of measurement techniques have shown that the PTA network is characterised by reticular strands, spaces, and regular hexagonal units smaller than vesicles, these observations prompting the formulation of a vesicle-network coincidence model of the presynaptic terminal. This model has been tested by tracing the profiles of vesicles within the PTA network and comparing their size and shape frequency distributions with those of osmicated synaptic vesicles. The distributions have been found to be essentially similar, suggesting that vesicles can be located within the network, and that the hexagonal network units are formed only in the presence of an underlying vesicular matrix.Additionally, the following points have emerged: 1) the dense projections in the two types of material appear to be equivalent; 2) a loose correlation exists between dense projections and vesicles in osmicated terminals, increase in the area of the dense projections being associated with a decrease in the area of the vesicles; 3) network and dense projection units are similar. In view of the similarity between network and dense projection units, the demonstrated vesicular basis of the network raises the question of whether dense projections are entirely independent structures, or whether they depend in part for their existence on the nearby presence of synaptic vesicles.This work was supported by the Arnold Yeldham and Mary Raine Research Foundation of the University of Western Australia and by the Australian Research Grants Committee and the Nuffield Foundation     

The hypothalamo-hypophysial system of the frog Rana temporaria

Summary The median eminence (ME) of the adult frog, Rana temporaria, was studied by means of electron microscopy including quantitative electron-microscopic autoradiography. In frogs captured in May and June numerous peptidergic neurosecretory fibres extending via the internal zone to the pars nervosa display large swellings containing few granules, mitochondria, neurotubules and cisternae of the smooth endoplasmic reticulum. In addition, few secretory globules up to 1.5 m in diameter occur in these varicosities. In animals collected during the autumn period many of these neurosecretory swellings filled with neurosecretory granules and polymorphic inclusions resemble Herring bodies. Three types of granule-containing neurosecretory fibres were observed in the external zone (EZ) of the ME of adult R. temporaria. Peptidergic A₁- and A₂-type fibres are characterized by granules 150–220 nm and 100–160 nm in diameter, respectively. Monoaminergic fibres of type B with granules approximately 100 nm in diameter represent 50% of all neurosecretory elements in the EZ of the frog ME; 12% of the total number of granule-bearing axons in the EZ actively taking up radiolabelled 5-hydroxytryptophan are thought to be serotoninergic terminals. Neurosecretory terminals of all types and glial vascular endfeet establish direct contacts with the perivascular space of the primary portal capillaries. Some neurosecretory terminals are separated from the lumen of the third ventricle by a thin cytoplasmic lamella of tanycytes. The possible physiological significance of this structural pattern is discussed.     

Nervous connections of the parietal eye in adult Lacerta s. sicula Rafinesque as demonstrated by anterograde and retrograde transport of horseradish peroxidase

Summary Subsequent to the injection of horseradish peroxidase into the parietal eye of adult Lacerta sicula, the course of the parietal nerve and its projections were determined.The parietal nerve enters the left habenular ganglion where it branches into a medial and a lateral route. Some nerve fibers decussate within the habenular commissure. Whereas this pathway exhibits a striking asymmetry at the level of the habenular ganglia, its projections to the dorsolateral nucleus of the thalamus, the periventricular hypothalamic area, the preoptic hypothalamic and telencephalic regions, and the pretectal area are arranged in a strictly symmetric manner. A possible innervation of tegmental areas could not be proven due to the presence of endogenous peroxidase within these regions. No parietal nerve fibers were observed in the optic tectum.In a few animals investigated, scattered labeled perikarya were located in the periventricular hypothalamic gray indicating a parietopetal innervation in Lacerta sicula. The injection of horseradish peroxidase into one of the lateral eyes revealed terminal areas of the optic nerve within the preoptic region, and the thalamic and pretectal nuclei, displaying partial overlapping with the projections of the parietal nerve to these areas.From the present investigation further evidence is obtained that the pineal complex of lower vertebrates is a component of the photoneuroendocrine system. Particular emphasis is placed upon the nervous connections between the parietal eye and the hypothalamus, described for the first time in the present study.Supported by the Deutsche Forschungsgemeinschaft (Grant Ko 758/1)In partial fulfillment of the requirements of the degree of Dr. med., Faculty of Medicine, Justus Liebig University of Giessen     

Photoreceptor cells and neural elements with long axonal processes in the pineal organ of the lamprey,Lampetra japonica,identified by use of the horseradish peroxidase method

Summary Horseradish peroxidase (HRP) was applied to the transected end of the pineal tract of the lamprey, Lampetra japonica. Distinct reaction products of HRP were observed in 2 types of cell other than ganglion cells. The first type of cell protrudes a knob-like process into the pineal lumen. This type of cell was clearly identified by electron microscopy as a photoreceptor cell; its outer segment was connected to the ellipsoid through a sensory cilium. The other type of cell was located among photoreceptor and supporting cells. The processes of these cells were thin and slender, and they obviously did not represent photoreceptor, supporting, or conventional ganglion cells. The present results indicate that, in the lamprey, some of the photoreceptor cells of the pineal organ project their axon-like processes toward the posterior commissure, but that there is also another type of cell displaying long axonal projections. HRP-containing cells were distributed randomly over the pineal organ and were occasionally also observed in the parapineal organ.     

On the Cytology and Fine Structure of the Neogregarine Syncystis aeshnae Tuzet and Manier, 1953 (Apicomplexa, Syncystidae)

The neogregarine Syncystis aeshnae Tuzet and Manier, 1953, was obtained from two species of dragonfly larvae, Aeshna grandis and Libellula quadrimaculata , collected in southern Sweden. Merogony, gamogony, and sporogony were observed in the adipose tissue, which was destroyed, and the haemocoele. Merozoites had a traditional apical complex. Gamonts associated in pairs did not unite conoid-to-conoid, and the border between gamonts was not found to break down. Each association produced 32 spores in bilobed gametocysts. Spores were elongate, oval, 4.0–5.8 × 8.0–10.0 μm in fixed and stained condition, each with 4+4 terminal and four equatorial thread-like projections. The thick-walled spore is considered the sporocyst. Species identification, ultrastructural cytology, and problems associated with using the terms "oocyst" and "sporocyst" are discussed.     

Central projections of sensory cells of the midleg of the locust,Schistocerca gregaria

The central projections of trichoid hairs and of some scolopidial organs of the mesothoracic leg of the locust Schistocerca gregaria were studied by using nickel chloride backfilling and single cell recording. Trichoid hair sensilla on different parts of the legs project somatotopically in the ventral part of the ipsilateral neuropile of the mesothoracic ganglion. Generally, distally located receptors have their terminal arborizations in ventro-lateral areas of the neuropile, and proximally located receptors in ventro-medial areas. The axons of the subgenual organ and tarsal chordotonal organs project into the intermediate neuropile.     

Projection areas and branching patterns of the tympanal receptor cells in migratory locusts,Locusta migratoria and Schistocerca gregaria

Summary In Locusta migratoria and Schistocerca gregaria, the projection areas and branching patterns of the tympanal receptor cells in the thoracic ganglia were revealed. Four auditory neuropiles can be distinguished on each side of the ventral cord, always located in the anterior part of the ring tract in each neuromere (two in the meta-, one in the meso-, and one in the prothoracic ganglion). Some of the receptor fibres ascend to the suboesophageal ganglion. There are distinct subdivisions within the auditory, frontal metathoracic and mesothoracic neuropiles. The arrangement of the terminal arborisations of the four types of tympanal receptor cells according to their different frequency-intensity responses is somatotopic and similar in the two ganglia. Here the receptor cells of type-1 form a restricted lateroventral arborisation. Cells of type-4 occupy the caudal part with a dorsorostral extension. Cells of type-2 and -3 arborise in a subdivision between both. Most of the stained low-frequency receptors (type-1, -2, and -3) terminate either in the metathoracic or, predominantly, in the mesothoracic ganglion. In contrast, the high-frequency cells (type-4) ascend to the prothoracic ganglion. The receptor fibres of the different types of receptor cells differ in diameter.Abbreviations aRT anterior part of the ring tract - cf characteristic frequency - MVT median ventral tract - SEG suboesophageal ganglion - SMC supramedian commissure - VMT ventral median tract - VIT ventral intermediate tract Supported by the Deutsche Forschungsgemeinschaft; part of program A7 in Sonderforschungsbereich 305 (Ecophysiology)     

Identification of serotonin- and vasopressin immunoreactivities in the suprachiasmatic nucleus of four mammalian species

Summary Cobalt fills from small, defined regions of the antenna in D. melanogaster show that the three types of sensilla on the third segment, the flagellum, and a fourth sensillum located in the arista, project into the glomeruli of the antennal lobe. We have identified 19 glomeruli in each lobe, according to their location, shape, and size. At least ten of these represent major projection areas of flagellar or aristal sensilla. The large majority of glomeruli is innervated from both antennae, but a small group of five receive exclusively ipsilateral input. A particular sensory fiber appears to terminate only in one specific glomerulus, either in the ipsilateral or in both lobes. Fills from flagellar regions bearing a single type of sensillum, yield a specific pattern of glomeruli containing stained terminals. Aristal projections remain strictly ipsilateral, whereas those from the other sensilla consist of an ipsilateral and a bilateral component. When filling from different points in an area bearing one type of sensillum, similar projections are produced, suggesting that projection patterns observed reflect predominantly the type of sensillum rather than its location on the flagellum. Accordingly, individual glomeruli might represent functional units, each receiving antennal input in a characteristic combination.We are indebted to Dr. H. Tobler for critical comments. R.F.S. was supported by the Swiss National Foundation (Grant No. 3.541-0.79) as well as a Travel Aid by the Swiss Academy of Sciences     

Specific Neurochemical Derangements of Brain Projecting Neurons in Apolipoprotein E-Deficient Mice

Abstract: Apolipoprotein E (apoE)-deficient mice provide a useful system for studying the role of apoE in neuronal maintenance and repair. Previous studies revealed specific memory impairments in these mice that are associated with presynaptic derangements in projecting forebrain cholinergic neurons. In the present study we examined whether dopaminergic, noradrenergic, and serotonergic projecting pathways of apoE-deficient mice are also affected and investigated the mechanisms that render them susceptible. The densities of nerve terminals of forebrain cholinergic projections were monitored histochemically by measurements of acetylcholinesterase activity, whereas those of the dopaminergic nigrostriatal pathway, the noradrenergic locus coeruleus cortical projection, and the raphe-cortical serotonergic tract were measured autoradiographically using radioligands that bind specifically to the respective presynaptic transporters of these neuronal tracts. The results obtained revealed that synaptic densities of cholinergic, noradrenergic, and serotonergic projections in specific brain regions of apoE-deficient mice are markedly lower than those of controls. Furthermore, the extent of presynaptic derangement within each of these tracts was found to be more pronounced the further away the nerve terminal is from its cell body. In contrast, the nerve terminal density of the dopaminergic neurons that project from the substantia nigra to the striatum was unaffected and was similar to that of the controls. The rank order of these presynaptic derangements at comparable distances from the respective cell bodies was found to be septohippocampal cholinergic > nucleus basalis cholinergic > locus coeruleus adrenergic > raphe serotonergic ? nigrostriatal dopaminergic, which interestingly is similar to that observed in Alzheimer's disease. These results suggest that two complementary factors determine the susceptibility of brain projecting neurons to apoE deficiency: pathway-specific differences and the distance of the nerve terminals from their cell body.     

The distribution of and interactions between GABA-immunoreactive and non-immunoreactive processes presynaptic to afferents from campaniform sensilla on the trochanter of the locust leg

Summary Campaniform sensilla on the trochanter of the mesothoracic legs of the locust were backfilled with cobalt salts or horseradish peroxidase for light and electron microscopy. The distribution of the terminal branches of afferent neurones in the thoracic ganglia were described from wholemount preparations and from thick slices through the ganglia. Ultrathin sections of identified branches were processed with GABA antibodies using a post-embedding immunogold technique and examined in the electron microscope. Input synapses were observed on fine varicose branches in all regions of the terminal arborisations close to the sites of afferent output. The major branches neither make nor receive synapses. Seventy-two percent of the input synapses are made by processes immunoreactive for GABA. Immunoreactive and non-immunoreactive processes synapse onto afferent terminals in close proximity. In some instances GABA-immunoreactive processes presynaptic to an afferent are also presynaptic to a non-immunoreactive presynaptic processes strongly suggesting that different presynaptic influences can interact directly with each other.     

Retinohypothalamic projections and immunocytochemical analysis of the suprachiasmatic region of the desert iguana Dipsosaurus dorsalis

Two separate and distinct retinal projections to the hypothalamus in the iguanid lizard Dipsosaurus dorsalis were described using horseradish peroxidase and cobalt-filling techniques. Both of the projections were unilateral and completely crossed; one terminated in the supraoptic nucleus and the other in the suprachiasmatic nucleus. Immunocytochemical analysis showed that the supraoptic nucleus contained cell bodies and fibers that cross-react with antibodies raised against arginine vasopressin, while the suprachiasmatic nucleus contained arginine vasopressin-like immunoreactive fibers emanating from cells in the nearby paraventricular nucleus. The suprachasmatic nucleus contained a dense plexus of fibers that cross-reacted with neuropeptide-Y antibody. Antiserum against vasoactive intestinal polypeptide showed no reactivity in any part of the forebrain, while antiserum against serotonin showed sparse and uniform reactivity throughout the forebrain, including the suprachiasmatic nucleus. These results, together with other data, indicate that the suprachiasmatic nucleus of D. dorsalis is homologous to the suprachiasmatic nuclei of rodents, structures known to contain circadian pacemakers. We suggest that the suprachiasmatic nucleus may play a similar role in the circadian system of D. dorsalis.     

Anterograde labelling from the optic nerve reveals multiple central targets in the teleost,Lethrinus chrysostomus (Perciformes)

Summary Cobaltous-lysine is transported anterogradely from the optic nerve of the teleost, Lethrinus chrysostomus (Lethrinidae, Perciformes). The marginal optic tract is labelled in longtitudinal bands of light and dark staining fibres which persists caudally within the ventral division but not in the dorsal division. This species possesses multiple central targets in the contralateral preoptic, diencephalic, pretectal, periventricular and tectal regions of the brain. In addition, a greater subdivision of the marginal optic tract is found to project to various nuclei. Ipsilateral projections are found in the suprachiasmatic nucleus and in the region of the horizontal commissure. Projections are also found in the telencephalic region of the nucleus olfactoretinalis and the thalamic region of the nucleus thalamoretinalis. The retinotopicity of some of these nuclei, found in previous studies, is discussed in relation to the possibility of specific sub-populations of retinal ganglion cells having different central targets.Abbreviations used in the Text and Figures A nucleus anteriorthalami - AO accessory optic nucleus - AOT accessory optic tract - AxOT axial optic tract - BO nucleus of the basal optic root - C cerebellum - HCv ventral division of horizontal commissure - I nucleus intermedius thalami - IL inferior lobe - MdOT medial optic tract - MO medulla oblongata - MOTd dorsal division of the marginal optic tract - MOTi intermediate division of the marginal optic tract - MOtv ventral division of the marginal optic tract - O olfactory bulb - OT optic tract - PC nucleus pretectalis centralis - PCo posterior commissure - Pd nucleus pretectalis dorsalis - PG preglomerular complex - PPd nucleus pretectalis periventricularis, pars dorsalis - PPv nucleus pretectalis periventricularis, pars ventralis - PSm nucleus pretectalis superficial pars magnocellularis - PSp nucleus pretectalis superficialis, pars parvocellularis - Sn suprachiasmatic nucleus - TEL telencephalon - TeO optic tectum - TL torus longtitudinalis - TrOlfR tractus olfactoretinalis - VCg granular layer of the valvula cerebelli - VCm molecular layer of the valvula cerebelli - VM nucleus medialis thalami - VL nucleus ventrolateralis thalami - VMdOT ventro-medial optic tract     

Neurotransmitters alter the numbers of synapses and organelles in photoreceptor terminals in the lamina of the housefly, Musca domestica

Various organelles in the lamina terminals of housefly photoreceptors exhibit daily rhythms having a circadian basis. These include changes in the numbers of photoreceptor tetrad and L2 feedback synapses, and longitudinal movements of screening pigment. Circadian information has previously been suggested to spread from the clock to the lamina via widefield cells expressing either 5-hydroxytryptamine or pigment-dispersing hormone-like immunoreactivity. We examined the action of these neuromodulators, and other candidate neurotransmitters, 4 h after injecting either the transmitter or a control into the medulla. We counted electron microscope profiles of organelles that normally exhibit circadian changes, and two types of invagination into photoreceptor terminals, capitate projections and inter-receptor invaginations. No single substance mediated the changes observed. Injected pigment-dispersing hormone peptide decreased the number of pigment granules, implicating this peptide in screening pigment migration, but produced no changes in synapse-related organelles. α-Aminobutyric acid exclusively decreased the number of L2 feedback synapses. Responses to other transmitters were specific, and often large, but generally not statistically significant. Histamine, for example, may decrease the number of tetrads, possibly by direct autoregulation. The results suggest that there is likely to be more than one effector in the circadian pathways to the lamina. Accepted: 11 August 1998     

Hygro- and thermoreceptors in tip-pore sensilla of the tarsal organ of the spider Cupiennius salei: innervation and central projection

The hygro- and thermoreceptive tarsal organ in the wandering spider Cupiennius salei is located on the tarsus of each walking leg and pedipalp, and consists of a tiny air-filled capsule in the cuticle. This capsule communicates with the outside world through a small aperture and contains seven nipple-shaped sensilla, each with a pore at its tip. In both their external morphology and internal structure, the sensilla are indistinguishable, although one sensillum is innervated by only two sensory cells, whereas the other six sensilla contain three sensory cells. Their dendrites are unbranched and terminate at the tip-pore, where they are enveloped by amorphous material that appears to limit their exposure to the atmosphere. Cobalt fillings reveal that each tarsal organ projects to three different areas within the suboesophageal ganglionic mass: (1) the sensory longitudinal tract 3 and 4; (2) the corresponding pedipalpal or leg ganglion; (3) a structured neuropil (here termed the Blumenthal neuropil) beneath the oesophagus. The multiple representation of sensory afferents from each tarsal organ in different regions of the suboesophageal ganglionic mass suggests parallel processing of hygro-/thermoreceptive information.     

Nuclear envelope projections from pronuclei in karyogamy ofSpirogyra

Karyogamy ofSpirogyra (S. verruculosa andSpirogyra sp.) was investigated by electron microscopy. After conjugation both male and female pronuclei migrated to the center of the zygote and adjoined. Many regular finger-like projections arose from the nuclear envelopes of the pronuclei; each involved both the outer and inner nuclear membranes and was ca. 0.17 μm in diameter. The inner membrane was underlaid by electron-dense bands parallel to one another in arrangement and perpendicular to the axis of protrusion in direction. Subsequently, the two pronuclei were connected by internuclear bridges, and the approximating surfaces of them were closely apposed. The bridges resembled the said projections both in diameter and structure, having a diameter of ca. 0.17 μm and electron-opaque bands lining the inner membrane. The adjacent surfaces of the two pronuclei approximated to each other more and more, and by 30 days after conjugation both pronuclei intermingled into a synkaryon, from which the projections disappeared. Probably, the nuclear envelope projections are involved in the initial contact between the two pronuclei and then the internuclear bridges are formed.