Putative neurohemal areas in the peripheral nervous system of an insect,Gryllus bimaculatus,revealed by immunocytochemistry

The morphology and position of putative neurohemal areas in the peripheral nervous system (ventral nerve cord and retrocerebral complex) of the cricket Gryllus bimaculatus are described. By using antisera to the amines dopamine, histamine, octopamine, and serotonin, and the neuropeptides crustacean cardioactive peptide, FMRFamide, leucokinin 1, and proctolin, an extensive system of varicose fibers has been detected throughout the nerves of all neuromeres, except for nerve 2 of the prothoracic ganglion. Immunoreactive varicose fibers occur mainly in a superficial position at the neurilemma, indicating neurosecretory storage and release of neuroactive compounds. The varicose fibers are projections from central or peripheral neurons that may extend over more than one segment. The peripheral fiber varicosities show segment-specific arrangements for each of the substances investigated. Immunoreactivity to histamine and octopamine is mainly found in the nerves of abdominal segments, whereas serotonin immunoreactivity is concentrated in subesophageal and terminal ganglion nerves. Immunoreactivity to FMRFamide and crustacean cardioactive peptide is widespread throughout all segments. Structures immunoreactive to leucokinin 1 are present in abdominal nerves, and proctolin immunostaining is found in the terminal ganglion and thoracic nerves. Codistribution of peripheral varicose fiber plexuses is regularly seen for amines and peptides, whereas the colocalization of substances in neurons has not been detected for any of the neuroactive compounds investigated. The varicose fiber system is regarded as complementary to the classical neurohemal organs.     

Development and sensitivity to serotonin of Drosophila serotonergic varicosities in the central nervous system

Serotonin is a classical small-molecule neurotransmitter with known effects on developmental processes. Previous studies have shown a developmental role for serotonin in the fly peripheral nervous system. In this study, we show that serotonin can modulate the development of serotonergic varicosities within the fly central nervous system. We have developed a system to examine the development of serotonergic varicosities in the larval CNS. We use this method to describe the normal serotonergic development in the A7 abdominal ganglion. From first to third instar larvae, the volume of the neuropil and number of serotonergic varicosities increase substantially while the varicosity density remains relatively constant. We hypothesize that serotonin is an autoregulator for serotonergic varicosity density. We tested the sensitivity of serotonergic varicosities to serotonin by adding neurotransmitter at various stages to isolated larval ventral nerve cords. Addition of excess exogenous serotonin decreases native varicosity density in older larvae, and these acute effects are reversible. The effects of serotonin appear to be selective for serotonergic varicosities, as dopaminergic and corazonergic varicosities remain qualitatively intact following serotonin application.     

Neuropeptide release from isolated myenteric nerve endings derived from the guinea pig myenteric plexus.

Isolated myenteric nerve varicosities prepared from the myenteric plexus of the guinea pig ileum were investigated as a suitable model system with which to study the release of several neuropeptide-like immunoreactivities (-LI). Basal release of substance P-LI, neurokinin A-LI, Leu-enkephalin-LI and Met-enkephalin-LI was determined, and clear depolarization-induced release of the enkephalin-LI's and neurokinin A-LI was obtained using this preparation, providing further support for their roles as putative mediators in the enteric nervous system. Evoked-release of these peptides was dependent on the presence in the incubation mixture of certain antagonists to known endogenous neuronal mediators. In the absence of such antagonists, no unequivocal evidence of release was seen. Clear evoked release of Leu-enkephalin-LI occurred only in the presence of the adenosine receptor antagonist 1,3-dipropyl-8-p-sulfophenylxanthine (DPSPX), atropine and naloxone. Release of Met-enkephalin-LI occurred in the presence of either atropine or naloxone. The release of neurokinin A-LI was evident in the presence of DPSPX. These findings suggest the existence of either distinct subpopulations of nerve varicosities or distinct neuronal pools containing each peptide and that these peptides may be under differential regulation by endogenous inhibitory mediators. It is concluded that, under suitable conditions, isolated myenteric nerve varicosities provide a useful model system for the study of release, and the modulation of release, of endogenous neuropeptides.     

Axonal varicosity density as an index of local neuronal interactions

Diffuse transmission is an important non-synaptic communication mode in the cerebral neocortex, in which neurotransmitters released from en passant varicosities interact with surrounding cells. In a previous study we have shown that the cholinergic axonal segments which were in the microproximity with dopaminergic fibers possessed a greater density of en passant varicosities compared to more distant segments, suggesting an activity-dependent level of en passant varicosities in the axonal zone of interaction. To further evaluate this plastic relationship, the density of cholinergic varicosities was quantified on fiber segments within the microproximity of activated or non-activated pyramidal cells of the prefrontal cortex (mPFC). Repetitive 14 days patterned visual stimulation paired with an electrical stimulation of the cholinergic fibers projecting to the mPFC from the HDB was performed to induce persistent axonal plastic changes. The c-Fos early gene immunoreactivity was used as a neuronal activity marker of layer V pyramidal cells, labelled with anti-glutamate transporter EAAC1. Cholinergic fibers were labeled with anti-ChAT (choline acetyltransferase) immunostaining. The density of ChAT+ varicosities on and the length of fiber segments within the 3 μm microproximity of c-Fos positive/negative pyramidal cells were evaluated on confocal images. More than 50% of the pyramidal cells in the mPFC were c-Fos immunoreactive. Density of ChAT+ varicosities was significantly increased within 3 μm vicinity of activated pyramidal cells (0.50±0.01 per μm of ChAT+ fiber length) compared to non-activated cells in this group (0.34±0.001; p≤0.05) or control rats (0.32±0.02; p≤0.05). Different types of stimulation (visual, HDB or visual/HDB) induced similar increase of the density of ChAT+ varicosities within microproximity of activated pyramidal cells. This study demonstrated at the subcellular level an activity-dependent enrichment of ChAT+ varicosities in the axonal zone of interaction with other neuronal elements.     

Role of nonsynaptic communication in regulating the immune response

The discovery of nonsynaptic communication in the 1960s and 1970s was an important milestone in investigating the function of the nervous system, and it revolutionized our view about information transmission between neurons. In addition, nonsynaptic communication has a practical importance not only within the nervous system, but in the communication between the peripheral nervous system and other organ systems. Nonsynaptic communication takes place in different immune organs, which are innervated by sympathetic nerve terminals. In addition, the function of microglia, one of the immunocompetent cell types of the brain, can also be affected by neurotransmitters released from axon varicosities. The various functions of immune cells are modulated by released neurotransmitters without any direct synaptic contact between nerve endings and targeted immune cells requiring only functional neurotransmitter receptors on immune cells. Here, we briefly overview the role of the various receptor subtypes mediating nonsynaptic modulation of the function of immunocompetent cells both in the periphery and in the central nervous system.     

Synaptic ultrastructure in nerve terminals of Drosophila larvae overexpressing the learning gene dunce

We investigated synaptic ultrastructure of individual nerve ending varicosities at the Drosophila larval neuromuscular junction in transgenic larvae overexpressing the learning gene dunce (dnc) in the nervous system. It was previously shown that cAMP is reduced to one-third normal in these larvae and that they have fewer nerve terminal varicosities and smaller junction potentials, although transmitter release from individual nerve ending varicosities is not significantly altered. We tested the hypothesis that synaptic ultrastructure is modified to compensate for possible reduced efficacy of synaptic transmission resulting from lower than normal cAMP. Synaptic size and number of presynaptic dense bodies (active zone structures) per synapse are modestly enhanced in transgenic larvae overexpressing the dnc gene product and in rutabaga (rut(1)) mutant larvae, which have reduced adenylyl cyclase activity and reduced neural cAMP. The incidence of complex synapses (possessing 2 or more presynaptic dense bodies) was not consistently different in experimental larvae compared to controls. The observations suggest that chronic reduction of cAMP levels in the nervous system of Drosophila larvae, although leading to a modest compensatory change in synaptic structure, does not markedly alter several synaptic ultrastructural parameters which are thought to influence the strength of transmitter release; thus, homeostatic mechanisms do not act to maintain normal-sized junction potentials by altering synaptic structure.     

Localization and distribution of catecholaminergic structures in the nervous system of Phocanema decipiens (nematoda)

The nervous system of Phocanema decipiens was examined with both the formaldeyhyde-induced and the glyoxylic acid fluorescence histochemical techniques. Green catecholaminergic structures were observed in 4 cephalic papillary nerves, 2 fibres with varicosities in the nerve ring as well as the ventral nerve cord and a pair of lateral nerves.The papillary nerves, extending from the nerve ring to the lips region, have cell bodies which are located anterior or adjacent to the nerve ring. Cell bodies of the lateral nerves are found within the lateral cord tissue posterior to the nerve ring. Each of these neurons has 3 processes—one joins with the nerve ring, the other merges with the ventral nerve cord and the third ends abruptly within the lateral cord.     

Microscopic anatomy and ultrastructure of the nervous system of <Emphasis Type="Italic">Phoronopsis harmeri</Emphasis> Pixell, 1912 (Lophophorata: Phoronida)

The microscopic anatomy and ultrastructure of the nervous system of Phoronopsis harmeri was investigated using histological techniques and electron microscopy. The collar nerve ring is basically formed by circular nerve fibers originating from sensitive cells of tentacles. The dorsal nerve plexus principally consists of large motor neurons. It is shown for the first time that the sensitive collar nerve ring immediately passes into the motor dorsal nerve plexus. The basic components of the nervous system have similar cytoarchitectonics and a layered structure. The first layer is formed by numerous nerve fibers surrounded by the processes of glia-like cells. The bodies of glia-like cells constitute the second layer. The third layer consists of neuron bodies overarched by the bodies of epidermal cells. The giant nervous fiber is accompanied by more than one hundred nerve fibers of a common structure and, thus, marks the true longitudinal nerve. The phoronids possess one or two longitudinal nerves. It is supposed that the plexus nature of the nervous system in phoronids may be related to their phylogenesis. A comparison of the nervous system organization and body plans among the Lophophorata suggests that the nervous system of phoronids cannot be considered as a reductive variant of the brachiopod nervous system. At the same time, the structure of the nervous system of bryozoans can be derived from that of phoronids.     

The Croonian Lecture 2000. Nicotinic acetylcholine receptor and the structural basis of fast synaptic transmission

Communication in the nervous system takes place at chemical and electrical synapses, where neurotransmitter-gated ion channels, such as the nicotinic acetylcholine (ACh) receptor, and gap junction channels control propagation of electrical signals from one cell to the next. Newly developed electron crystallographic methods have revealed the structures of these channels trapped in open as well as closed states, suggesting how they work. The ACh receptor has large vestibules extending from the membrane which shape the ACh-binding pockets and facilitate selective transport of cations across a narrow membrane-spanning pore. When ACh enters the pockets it triggers a concerted conformational change that opens the pore by destabilizing a gate in the middle of the membrane made by a ring of pore-lining alpha-helical segmets. The alternative 'open' configuration of pore-lining segments reshapes the lumen and creates new surfaces, allowing the ions to pass through. The gap junction channel uses a similar structural mechanism, involving coordinated rearrangements of alpha-helical segments in the plane of the membrane, to open its pore.     

Adrenergic innervation of the arteries of different diameters in the human and animal pia mater

The adrenergic nervous plexuses of the pial arteries from 450- to 50 micron in diameter have been studied in dogs, cats and humans from 4 age groups (22-44 years, 55-64 years, 65-74 years and 75-86 years old). It has been found that the decrease in the vessel diameter was accompanied by a marked decline in the absolute number of nervous fibers in the nervous plexuses, however the concentration of the nerve fibers has not revealed any significant differences between human arteries from 450 to 100 micron in diameter and animal arteries from 300 to 80 micron in diameter. The number of varicosities-thickness along the nerve fiber--was the greatest in 200-100 micron human arteries and in 80-60 micron animal arteries. With ageing, the number of varicosities in the adrenergic nervous plexus of human pial vessels decreased faster than in the nerve fibers.     

Fine structure and innervation of the avian adrenal gland

Summary Apart from cholinergic nerve fibers, which make up the main part of efferent fibers to the avian adrenal gland (Unsicker, 1973b), adrenergic, purinergic and afferent nerve fibers occur. Adrenergic nerve fibers are much more rare than cholinergic fibers. With the Falck-Hillarp fluorescence method they can be demonstrated in the capsule of the gland, in the pericapsular tissue and near blood vessels. By their green fluorescent varicosities they may be distinguished characteristically from undulating yellow fluorescent ramifications of small nerve cells which are found in the ganglia of the adrenal gland and below the capsule. The varicosities of adrenergic axons exhibit small (450 to 700 Å in diameter) and large (900 to 1300 Å in diameter) granular vesicles with a dense core which is usually situated excentrically. After the application of 6-hydroxydopamine degenerative changes appear in the varicosities. Adrenergic axons are not confined to blood vessels but can be found as well in close proximity of chromaffin cells. Probably adrenergic fibers are the axons of large ganglion cells which are situated mainly within the ganglia of the adrenal gland and in the periphery of the organ and whose dendritic endings show small granular vesicles after treatment with 6-OHDA.A third type of nerve fiber is characterized by varicosities containing dense-cored vesicles with a thin light halo, the mean diameter (1250 Å) of which exceeds that of the morphologically similar granular vesicles in cholinergic synapses. Those fibers resemble neurosecretory and purinergic axons and are therefore called p-type fibers. They cannot be stained with chromalum-hematoxyline-phloxine. Axon dilations showing aggregates of mitochondria, myelin bodies and dense-cored vesicles of different shape and diameter are considered to be afferent nerve endings. Blood vessels in the capsule of the gland are innervated by both cholinergic and adrenergic fibers.Supported by a grant from the Deutsche Forschungsgemeinschaft (Un 34/1).     

Synaptic function in the nervous system: A theory and its application

The objective of this paper is to present a new theory of synaptic function in the nervous system. The basis for this theory is the experimental demonstration that a nerve impulse assumes five different forms as it advances through the synaptic region, and that five basic mathematical operations have been identified as being involved in the transformation of one form into another form. As a result of these data, the synaptic region is regarded as a functional unit where information coming to it is unpacked, processed, stored, and retrieved for transit to another synaptic region or effector site. The data also suggests that a nerve impulse is a bolus of energy, therefore, without substance; that it contains information coded in its shape or form; that it is precisely described mathematically. Furthermore, the data suggests synaptic regions process these nerve impulses by applying mathematical operations to them; that function in the synaptic region is highly stereotyped (programmed); that chemical substances are associated with the mathematical operations. The basic approach of this theory is to regard a significant portion of the nervous system as an interface between the external universe and man himself. As an interface, the nervous system receives and processes information from both the external universe and man himself in a programmed manner. The interface functions by converting the information it receives into a bolus of energy, the nerve impulse, then processes the bolus by converting it into numbers or functions and applying mathematical operation to it.     

NERVOUS ORGANIZATION AND THE PROBLEM OF THE SYNAPSE IN ACTINIA EQUINA

The nervous system of Actinia equina was studied by routinehistological methods and by metallic impregnation techniques.Some preliminary results from electron microscopy are included. The organization of the nervous system of this species is morecomplex than that of other anthozoans; it consists of two interconnectednerve plexuses which are developed to differing degrees in variousparts of the body. These are: (1) a superficial (outer) plexuslying in the ectoderm, and (2) a deeper (inner) plexus constitutingthe main nerve net, lying in the mesoderm. The former is composedof bipolar and multipolar nerve cells, and the latter of multipolarcells. Receptor cells in the ectoderm make contact with fibersof the ectodermal plexus. Processes from the mesodermal plexusrun out to the muscle fibers. Connections between the receptor cells and the nerve processesof the superficial plexus and between the processes of the cteeperplexus and the muscle fibers appear to be of the discontinuous(synaptic) type. In the nerve nets themselves, although someconnections resembling synapses have been seen, most of thenerve elements stand in direct connection with one another,so that the system must be regarded as at least partly syncytial.Evidence is given for the growth of the nerve net, in step withthe general growth of the animal, by division of nuclei followedby their movement apart within the syncytium. The distribution of the nerve elements in various parts of thebody, the interconnections between these regions, and the cytologicalcharacteristics of the cells are described. Ways in which excitationcould pass from one part to another are discussed.     

Efferent fibers and daily rhabdomal changes in the anteromedial eye of the liphistiid spider,Heptathela kimurai

The presence of efferent fibers in the retina of liphistiid spiders, kept in natural daily cycles of illuminance, was examined by electron microscopy. The efferent fibers were observed to extend their processes through the ocellar nerve to the retina. They contained characteristic large electron-dense granules and branched repeatedly within the retina with varicosities, to provide synaptoid contacts with the receptor cells. They ran mostly among receptor cells and glial cells but sometimes protruded into receptor cells to establish invaginated synaptoid contacts. The synaptoid structures were characterized by spherical clear vesicles located at the presynaptic region, with electron-dense material adhering to the plasma membranes of the receptor cell and the efferent fiber, and a cleft about 10 nm wide formed by the two opposed parallel membranes. The clear vesicles and the electron-dense granules were secreted by exocytosis. The efferent fiber was characteristically presynaptic in relation to the receptor cell. In addition, the rhabdoms differed in size from day to night.     

Cutaneous and subcutaneous sensory receptors of the hagfish Myxine glutinosa with special respect to the trigeminal system

The integument of the hagfish Myxine glutinosa is described with respect to the topography and the fine structural organization of the dermal and hypodermal nerve fiber plexus. Both nerve fiber plexuses contain small ganglion cells with axodendritic and axosomatic synapscs. The six barbels of the head (4 nasal and 2 oral barbels) are supplied with about 5600 afferent trigeminal nerve fibers via the right and left ophthalmic nerve. With respect to the topography of the sensory nerve terminals in the barbels different types of receptors are termed the external cuff receptor, internal cuff receptor, and perichondrial receptor. Free nerve terminals occur within the epidermal layer, especially at the tip region of the barbels and in the glassy membrane of the dermis. The hypodermal edge receptor organ extends from the ventral nasal barbel to the oral barbel. A mechanoreceptive function of the different receptor types is discussed. The innervation pattern of the barbel is similar to the innervation of the mammalian sinus hair. In this context, the barbel is a highly differentiated receptor organ able to explore the nearest surroundings with high stereognostic perception. The ganglion cells of the skin seem to represent a part of the peripheral autonomic nervous system, which is involved in the control of secretion mechanisms.     

Neuropeptide (neuropeptide Y, neurotensin, vasoactive intestinal polypeptide, substance P, calcitonin gene-related peptide, somatostatin) immunohistochemistry and ultrastructure of renal nerves

With the use of several region-specific antisera and the peroxidase-antiperoxidase (PAP) technique, several regulatory polypeptides were localized in nerves of the kidney. Neuropeptide Y (NPY)- immunoreactivity (IR), neurotensin (NT)-IR and vasoactive intestinal polypeptide (VIP)-IR occurred at high densities in all segments of the renal arterial system forming a perivascular plexus. Furthermore, NT-IR nerves were particularly frequent at the juxtaglomerular apparatus (JGA). Calcitonin gene-related peptide (CGRP)-IR was mainly concentrated in nerves supplying the hilus arteries and the JGA. Substance P (SP)-IR was predominantly found in large varicosities close to large renal arterial vessels and in the vicinity of the JGA. Somatostatin (SOM)-IR was only observed in single varicosities located at the media-adventitia border of large renal hilus arteries. The peptidergic nerves are correlated to their ultrastructural counterpart. In addition, the distribution patterns and the frequency of the different types of renal peptidergic nerve fibres are evaluated and compared. The functional role of these neuropeptides and their origin within the efferent branch of this part of the peripheral autonomic nervous system is discussed. Furthermore, the implication of some of the neuropeptides studied in afferent renal innervation is also substantiated.     

Close anatomical relationships between nerve fibers and MHC class II-expressing dendritic cells in the rat liver and extrahepatic bile duct

 The anatomical relationships between immunocytochemically identified nerve fibers and MHC class II-expressing antigen presenting dendritic cells were investigated in the rat hepatobiliary system using immunocytochemistry, confocal laser scanning, and electron microscopy. Close proximity of nerve fiber varicosities immunostained for PGP 9.5 and MHC class II-expressing dendritic cells was frequently observed in the wall of extrahepatic bile ducts, in Glisson’s area, around central and hepatic veins, and in the liver capsule. Contacts between nerve fibers staining for substance P, calcitonin gene-related peptide, calretinin, and vasoactive intestinal polypeptide and dendritic cells were more often observed around extrahepatic bile ducts than in Glisson’s area. Nerve fibers immunostaining for tyrosine hydroxylase and neuropeptide Y were numerous both in the wall of extrahepatic bile ducts and in Glisson’s area and frequently contacted dendritic cells there. At the ultrastructural level, close membrane contacts between bare axolemmal areas of unmyelinated nerve fibers and processes of MHC class II-expressing cells were observed. These results demonstrate close anatomical relationships of nerve fibers from various sources with antigen presenting dendritic cells in the visceral domain and suggest modulation of antigen presentation by the autonomic nervous system. Accepted: 29 September 1997     

Development of P2X receptor clusters on smooth muscle cells in relation to nerve varicosities in the rat urinary bladder

Postnatal development of the distribution of different isoforms of purinergic (P2X) receptors on smooth muscle cells in relation to the development of the innervation of the cells by nerve varicosities in the rat urinary bladder has been determined with immunofluorescence and confocal microscopy. Antibodies against the extracellular domains of the P2X₁ to P2X₆ receptors were used to detect the receptors in the bladder. Several other antibodies were used to identify sympathetic varicosities and Schwann cells. At one day postnatal (D1) there were few strings of varicosities denoting isolated axons, with most axons confined to large nerve trunks. Small size clusters of P2X₁ to P2X₆ receptor subtypes (about 0.4 µm diameter) were observed in the muscle which were independent of each other, and sometimes juxtaposed to the rare isolated varicosity strings. At D4 large numbers of strings of varicosities could be discerned throughout the detrusor. Most of these clouds of small P2X₁ to P2X₆ receptor clusters in their immediate vicinity. Some of these were colocalised with the varicosities, which were of parasympathetic origin as they failed to counter-stain with antibodies to tyrosine hydroxylase. Up to D14 there was a gradual coalescence of many of the isolated P2X_1–6 small receptor clusters so that they became colocalised, often at varicosities. Most of the varicosities in isolated strings possessed receptor clusters at this time. By D21 it was rare to find varicosity strings in the detrusor that were not either in close juxtaposition with P2X small receptor clusters or possessing such clusters in colocalisation. However, large numbers of small P2X receptor clusters, many of which consisted of a mixture of isoforms, could be found spatially unrelated to nerve varicosities throughout the detrusor muscle. In the adult, single axons were either coextensive with one or more isoforms of P2X receptor clusters or these were immediately juxtaposed to the axons so that is was rare to find a varicosity that did not possess a receptor cluster. However, different combinations of colocalised P2X receptor isoforms could still be discerned in small clusters unrelated to varicosities. These observations are discussed in relation to the mechanism of formation of the receptor clusters and their migration beneath parasympathetic varicosities during development.