Some aspects of the structural organization of the arthropod ganglia

Summary This paper deals with the fine structure of the abdominal ganglia of several species of arthropods belonging to the classes Arachnida, Crustacea, Myriapoda and Insecta. The tissues were fixed in osmium tetroxide and embedded in n-butyl methacrylate or fixed in potasium permanganate and embedded in a mixture of X 133/2097 and Araldite.A comparative study was made in order to discriminate between those structural characteristics of the nervous system appearing only in determined taxonomic groups and those belonging to a fundamental plan common to the whole Phylum. This work covers the morphology of neurons, glial cells, neuropilic nerve fibers and neuronal connections.Most arthropod neurons are pear-shaped with only one prolongation and the nucleus is located in the center of the soma, enveloped by two membranes showing numerous pores. Cisternae of the ER have frequently been observed in continuity with this nuclear envelope. After osmic fixation the nuclear content appears to consist of small dense granules distributed at random in the nucleoplasm. In addition to these small perticles there are, in some species, large chromatin blocks. The use of Permanganate as fixative introduces important changes in the nuclear aspect; most of the nuclei look washed and the nuclear content acquires an homogeneous appearance.The cytoplasm of the neurons contains a complex system of internal membranes consisting of cisternae and tubuli of the ER system, lamellae of the Golgi complex and invaginations of the plasma membrane. In most species the elements of the ER system are distributed at random in the cytoplasm but in the neurons of Bothriurus bonariensis there are parallel aggregations of membranes similar to the Nissl bodies found in vertebrates.It was found in some of the species studied (Armadillidium vulgare and Lithobius Sp.) that the internal membrane system of the nerve cells is mainly represented by Golgi elements while the ER system seems to be poorly developed.Besides the membranous components, the neuronal cytoplasm contains mitochondria, multivesicular bodies and dense granules of neurosecretory material.Neuroglial cells are mainly characterized by their nuclear structure. After the action of osmium tetroxide, glial nuclei show irregular masses of chromatin inmersed in a nucleoplasm of low electron density. In permanganate fixed material these chromatin blocks appear as blank spaces.In the cytoplasm of these cells there are mitochondria, membranes pertaining to the ER system and elements of the Golgi complex but in some of the species studied gliofibrils and granules of pigment were found.Three main types of neuroglial cells have been recognized in an arthropod ganglia. These are: subcapsular glial cells, neuron satellites and nerve fiber satellites.The neuropile occupies the central region of the ganglion and consists of a great number of nerve fibers intermingled with glial processes. The neuropilic n. fibers consistently show profiles of ER membranes and tubuli pertaining to the ER system. In some of these fibers the ER reaches a high degree of development. In Armadillidium there is a special type of n. fiber containing a regular sequence of transversally oriented cisternae. Arthropod fibers sometimes contain thin parallel filaments as well as typical ER elements.Mitochondria, small vesicles and dense granules are commonly found within the neuroplasm of the neuropilic fibers. It is important to note that in arthropods, microvesicles are not restricted to the terminal region of the nerve fibers but that they may also occur all along the fibers.Arthropod neurons are enveloped by a glial insulating capsule and therefore interneuron contacts may only occur at neuropile level. These contacts are of three different morphological types: cross contacts, longitudinal contacts and end-knob contacts. At the level of longitudinal and cross contacts the neuroplasm shows no increase in the number of microvesicles or mitochondria. In the end-knob contacts, on the contrary, large numbers of microvesicles appear concentrated in the pre-synaptic fiber only, and occasionally in both fibers the pre-synaptic and the post-synaptic.It is maintained that funcional interneuron connections may result not only from contacts between fibers containing vesicles, but also between fibers in which vesicles are absent.     

Postembryonic development of leucokinin I-immunoreactive neurons innervating a neurohemal organ in the turnip moth Agrotis segetum

Summary In the abdominal ganglia of the turnip moth Agrotis segetum, an antibody against the cockroach neuropeptide leucokinin I recognizes neurons with varicose fibers and terminals innervating the perisympathetic neurohemal organs. In the larva, the abdominal perisympathetic organs consist of a segmental series of discrete neurohemal swellings on the dorsal unpaired nerve and the transverse nerves originating at its bifurcation. These neurohemal structures are innervated by varicose terminals of leucokinin I-immunoreactive (LKIR) fibers originating from neuronal cell bodies located in the preceding segment. In the adult, the abdominal segmental neurohemal units are more or less fused into a plexus that extends over almost the whole abdominal nerve cord. The adult plexus consists of peripheral nerve branches and superficial nerve fibers beneath the basal lamina of the neural sheath of the nerve cord. During metamorphosis, the LKIR fibers closely follow the restructuration of the perisympathetic organs. In both larvae and adults the LKIR fibers in the neurohemal structures originate from the same cell bodies, which are distributed as ventrolateral bilateral pairs in all abdominal ganglia. The transformation of the series of separated and relatively simple larval neurohemal organs into the larger, continuous and more complex adult neurohemal areas occurs during the first of the two weeks of pupal life. The efferent abdominal LKIR neurons of the moth Agrotis segetum thus belong to the class of larval neurons which persist into adult life with substantial peripheral reorganization occurring during metamorphosis.     

On the issue of innervation of dorsal longitudinal musculature of <Emphasis Type="Italic">Lymnaea stagnalis</Emphasis> with inferior cervical nerve

Using the method of the anterograde dextran tetramethylrhodamin transport, there is obtained the topographic picture of branching of inferior cervical nerve axons on fibers of the dorsal longitudinal muscle in Lymnaea stagnalis (L.). Using the retrograde staining, the neuronal bodies sending their processes into this nerve are marked. Manifestations of asymmetry in distribution of neurons stained through the right and left nerves are described. The electron microscopic studies have shown that the main number of the inferior cervical nerve axons is represented by thin fibers presumably belonging to the sensory cells. A part of the nerve fibers and their endings show imunoreactivity to serotonin and acetylcholine. The serotoninergic fibers predominate quantitatively over the cholinergic ones and account for a half of the fibers stained with dextran. A possible functional role of the serotoninergic and cholinergic innervation of the dorsal longitudinal muscle in Lymnaea stagnalis is discussed.Translated from Zhurnal Evolyutsionnoi Biokhimii i Fiziologii, Vol. 40, No. 6, 2004, pp. 569–578.     

Serotonin-like immunoreactivity in the central nervous system and gonad of the scallop,Patinopecten yessoensis

Summary The localization of neurons containing serotonin in the central nervous system and the gonad of the scallop, Patinopecten yessoensis, was examined immunohistochemically. In the central nervous system a large number of immunoreactive perikarya were observed in the following regions: a part of the anterior lobe of the cerebral ganglion; the posterior lobe of the cerebral ganglion; the pedal ganglion; and the accessory ganglion. No immunoreactive perikarya were found in the visceral ganglion. Numerous immunoreactive fibers were revealed in the neuropil of all central ganglia. In the gonadal region immunoreactive fibers were distributed around the gonoduct and along the germinal epithelium.This work was supported by a grant from the Ministry of Education, Science and Culture, Japan     

Effect of vasopressin on neuronal responses of theNucl. Caudalis of the spinal trigeminal tract in rats

Effect of vasopressin on responses of individual neurons of thenucl. caudalis of the spinal trigeminal tract was studied on rats under urethan anesthesia; the responses were evoked by nociceptive (stimulation of the tooth pulp) or non-nociceptive (stimulation of Aa fibers of the infraorbital nerve) afferent activation. After injection of 10 nM vasopressin into the recording zone, responses evoked by stimulation of the tooth pulp were suppressed in all studied neurons of the high-threshold group; the same was true as to responses induced by stimulation of the tooth pulp and infraorbital nerve in most neurons of the convergent group. At the same time, vasopressin did not change the responses evoked by stimulation of Aa fibers of the infraorbital nerve in neurons of the low-threshold group. Possible involvement of vasopressin in the process of pain suppression is discussed.     

Glutamic acid decarboxylase (GAD)-like immunoreactivity in the pedal ganglion of Mytilus galloprovincialis

Summary A substance immunologically related to vertebrate glutamic acid decarboxylase (GAD) has been visualized in the pedal ganglion of Mytilus with the pre-embedding peroxidase-antiperoxidase method, by use of an antiserum raised in sheep against rat brain GAD. The results show that GAD-like immunoreactivity is present both in neuronal perikarya and in nerve fibers. Positive neurons are located mainly among the fibers of the ganglion neuropil at the commissural level, and more rarely close to unreactive cortical cell bodies. Immunoreactive nerve fibers are observed throughout the neuropil and also in cerebropedal and pedal nerves.Supported by Ministero Pubblica Istruzione (40%)     

Neurosecretory cells in the circadian-clock system of the scorpion,Androctonus australis

Summary The somata of the efferent neurosecretory fibers that control the circadian sensitivity rhythm in the median eyes of the scorpion, Androctonus australis, were detected in the brain by retrograde labeling with Lucifer Yellow CH. A total of 20–40 neurons are arranged in two groups displaying a bilaterally symmetrical, marginal position near the circumesophageal connectives. Half the cells in each group send fibers into the ipsilateral optic nerve; the fibers from the other half enter the contralateral optic nerve.     

Selective uptake of noradrenaline,dopa, and 5-Hydroxytryptamine by the nervous system of Phocanema decipiens (Nematoda): A light autoradiographic and ultrastructural study

As determined by light microscopic autoradiography, parts of the nervous system of Phocanema decipiens have selective and high affinity mechanisms for the uptake of tritiated noradrenaline, dihydroxyphenylalanine (dopa) and 5-hydroxytryptamine. In the nervous system, noradrenaline is accumulated only by the four papillary nerves and two fibers in the nerve ring. The precursor dopa is also taken up by these neurons and, in addition, by the lateral nerves. 5-Hydroxytryptamine is accumulated by the three pharyngeal nerves, two cells in each lateral ganglion, and two other fibers in the nerve ring. With adjacent ultrathin sections, the labelled papillary nerve and lateral ganglion were examined ultrastructurally and found to contain various dense core vesicles which are similar to those in other aminergic neurons. The adjoining unlabelled cells of the same neurons are found, on the other hand, to contain dense agranular vesicles. With these results, the noradrenaline accumulating neurons are suggested to be noradrenergic and to contain the amine synthesizing enzymes. The lateral nerves are regarded, for the present, as dopaminergic neurons. These suggestions are in agreement with the previous demonstration of catecholaminergic neurons in this nematode. The 5-hydroxytryptamine accumulating neurons are tentatively identified as tryptaminergic.     

Immunocytochemical localization of a substance in the eyestalk of the prawn,Palaemon serratus,reactive with an anti-FMRF-amide rabbit serum

Summary By use of a specific antiserum against the molluscan cardio-excitatory tetrapeptide FMRF-amide in combination with the PAP-method it was possible to obtain positive immunocytochemical reactions in several neurosecretory regions of the eyestalk of the prawn Palaemon serratus. FMRF-amide-like material was found in perikarya and nerve fibers of the medulla terminalis and in neurons in the lamina ganglionaris. The immunoreactivity observed in the glandular tissue located at the basal insertion of the eyestalk muscles must be ascribed to a non-specific reaction. The identification of immunopositive nerve fibers, ending on a nerve bundle in the medulla terminalis, and the fact that immunoreactive material was absent in the neurohemal sinus gland seem to indicate a neurotransmitter/neuromodulator function.     

Anatomy of neurons crossing the tritocerebral commissures of the cockroach Periplaneta americana (Blattaria)

The neuronal connections of the tritocerebral commissures of Periplaneta americana were studied in the brain-suboesophageal ganglion complex and the stomatogastric nervous system by means of heavy metal iontophoresis through cut nerve ends followed by silver intensification. The tritocerebral commissure 1 (Tc1) contains mainly the processes of the subpharyngeal nerve (Spn) whose neurons are located in both tritocerebral lobes and in the frontal ganglion. Some neurons of the frontal ganglion project through the Tc1 to the contralateral tritocerebrum. A few fibers in this commissure were observed projecting to the protocerebrum and the suboesophageal ganglion. There are tritocerebral neurons which pass through the Tc1 or the tritocerebral commissure 2 (Tc2) and extend on into the stomatogastric nervous system. One axon of a descending gaint neuron appears in the Tc2. This neuron lies in the tritocerebrum and connects the brain to the contralateral side of the ventral nerve cord. In addition, sensory fibers of the labral nerve (Ln) traverse both commissures to the opposite tritocerebrum. The anatomical and physiological relevance of the identified neuronal pathways is discussed. © 1995 Wiley-Liss, Inc.     

Met5-enkephalin-Arg6-Gly7-Leu8-immunoreactive nerve fibers in the major salivary glands of the rat: evidence for both sympathetic and parasympathetic origin

Summary The localization of the proenkephalin A-derived octapeptide, Met⁵-enkephalin-Arg⁶-Gly⁷-Leu⁸ (MEAGL), was studied in the major salivary glands of Sprague-Dawley and Wistar rats with the indirect immunofluorescence method. MEAGL-immunoreactive nerve fibers were found around the acini, along intra-and interlobular salivary ducts and in close contact with blood vessels. In the parotid and submandibular glands tyrosine hydroxylase (TH) immunoreactivity was observed in nerve fibers around the acini, in association with intra- and interlobular salivary ducts and around blood vessels, while in the sublingual gland TH-immunoreactive nerve fibers were only seen around blood vessels. Parasympathetic neurons in submandibular ganglia contained MEAGL immunoreactivity. Moderate TH immunoreactivity was seen in some neurons of the submandibular ganglia. A subpopulation of sympathetic principal neurons in the superior cervical ganglion were immunoreactive for both MEAGL and TH. In the trigeminal ganglion, no MEAGL-immunoreactive sensory neurons or nerve fibers were observed. Superior cervical ganglionectomies resulted in a complete disappearance of TH-immunoreactive nerve fibers, while MEAGL-immunoreative nerve fibers were still present in the glands. The presence of MEAGL immunoreactivity in neurons of both sympathetic superior cervical ganglia and parasympathetic submandibular ganglia and the results of superior cervical ganglionectomies suggest, that MEAGL-immunoreactive nerve fibers in the major salivary glands of the rat have both sympathetic and parasympathetic origin.     

The localization of the beta-subtype of protein kinase C (PKC-beta) in rat sympathetic neurons

The localization of PKC-beta was studied in rat sympathetic neurons using a polyclonal antibody specific for the beta 1- and beta 2-subspecies. The tissues studied included the superior cervical (SCG) and hypogastric (HGG) ganglia and the target tissues of the SCG and HGG neurons: the submandibular gland, iris, prostate and vas deferens. PKC-beta-LI was found in nerve fibers in both ganglia. A proportion of the fibers in the SCG disappeared after decentralization, suggesting that the fibers were of both pre- and postganglionic origin. The somata of the HGG and SCG neurons expressed varying amounts of PKC-beta-LI, the majority of SCG neurons being labelled only after colchicine treatment. In all target tissues there were PKC-beta-immunoreactive nerve fibers in bundles, but the most peripheral branches of the fibers were negatively labelled. The results show that PKC-beta-LI is widely present in sympathetic postganglionic neurons with mainly quantitative differences. The lack of PKC-beta in the most peripheral branches of nerve fibers might be a general feature of sympathetic postganglionic neurons, suggesting that the participation of PKC-beta in neurotransmitter release and in other functions in nerve terminals in sympathetic adrenergic neurons is unlikely.     

Ultrastructure of the circumoral nerve ring and the radial nerve cords in holothurians (Echinodermata)

The circumoral nerve ring and the radial nerve cords (RNCs) of Eupentacta fraudatrix and Pseudocnus lubricus (Holothuroidea) were examined as an example of holothurian nervous tissue. The RNC is composed of outer ectoneural and inner hyponeural layers, which are interconnected with one another via short neural bridges. The circumoral nerve ring is purely ectoneural. Both ectoneural and hyponeural components are epithelial tubes with a thick neuroepithelium at one side. A thin ciliated non-neuronal epithelium complements the neuroepithelium to form a tube, thereby enclosing the epineural and hyponeural canals. The whole of the ectoneural and hyponeural subsystems is separated from the surrounding tissue by a continuous basal lamina. The nerve ring and the ectoneural and hyponeural parts of the radial nerves are all neuroepithelia composed of supporting cells and neurons. Supporting cells are interpreted as being glial cells. Based on ultrastructural characters, three types of neurons can be distinguished: (1) putative primary sensory neurons, whose cilium protrudes into the epineural or hyponeural canal; (2) non-ciliated neurons with swollen rough endoplasmic reticulum cisternae; (3) monociliated neurons that are embedded in the trunk of nerve fibers. Different types of synapses occur in the neuropile area. They meet all morphological criteria of classical chemical synapses. Vacuolated cells occur in the neuroepithelium of E. fraudatrix, but are absent in P. lubricus; their function is unknown. The cells of the non-neuronal epithelia that overlie the ectoneural and hyponeural canals are hypothesized to belong to the same cell type as the supporting cells of the neuroepithelium.     

The distribution of an immunoreactive endothelin-like substance in the nervous system of the nereid,Neanthes diversicolor (Annelida)

Summary The distribution of an immunoreactive endothelin-1-like peptide was investigated in the nereid, Neanthes diversicolor, using an antiserum raised against synthetic endothelin-1. Immunoreactive perikarya were localized in the brain, and nerve fibers containing endothelin-1-like material were found in the neuropil occupying the central portion of the brain. No immunostained fiber elements were traced in the circumesophageal connectives. Immunoreactive perikarya occurred in the subesophageal ganglion. From this ganglion, specifically stained fibers run posteriorly toward the ventral nerve cord. In each segmental ganglion, immunoreactive neurons were observed in medio-ventral and latero-ventral regions, and one or two marked fibers extended to the parapodium. In the parapodium, small immunoreactive perikarya and fiber elements were visible. Immunolabeled fibers occurred in the stomatogastric nerves, in the wall of the buccal cavity, and in the pharynx, esophagus, intestine and its anal region. Immunoreactive perikarya and nerve fibers were visualized between the circular muscle layer and epithelial cell layer in the esophagus and intestine. The endothelin-1-like substance shown to occur in N. diversicolor appears to function as a neurotransmitter or neuromodulator.     

Multisegmental cobalt filling of the dorsal giant fibers in the nervous system of the earthworm,Lumbricus terrestris

Summary The anatomical organization of the two dorsal giant fiber systems of the earthworm Lumbricus terrestris is demonstrated in whole mounts and serial-section reconstructions based on backfillings of the ventral nerve cord with cobalt chloride. Both the medial and lateral fiber systems can be labeled selectively over more than ten body segments. They show a characteristic segmental pattern of collaterals with some modification in tail segments and of dorsal plasma protrusions in the unpaired medial giant fiber presumably representing openings in the myelin sheath. We found no multisegmental cobalt transport in other large neurons of the nerve cord. Cobalt passes through the segmentai septa between consecutive axonal elements of the metameric giant fibers and presumably also through commissural contacts between specific collaterals of the lateral giant fibers. Since these sites of contact are known to represent electrical synapses, cobalt coupling may, in L. terrestris, correlate with functional electrotonic coupling.Abbreviations CL collateral of lateral giant fiber - CM collateral of medial giant fiber - GIN giant interneuron - LGF lateral giant fiber - MGF medial giant fiber - SN segmental nerve     

Effects of temperature stress on NO-synthase and tyrosine hydroxylase activities in the central nervous system of bivalve molluscs

Effects of temperature stress on activities of NO-synthase (NOS) and tyrosine hydroxylase (TH) in the CNS of two species of bivalve molluscs, Mizuchopecten yessoensis and Chlamys farreri nipponensis (Pectinidae) were studied using NADPH-diaphorase histochemistry and immunocytochemistry. General and specific peculiarities in distribution and relative proportion of TH- and NO-containing neurons in the CNS nerve ganglia were revealed in norm and under stress at 30°C for 10, 30, and 60 min. The initial stress stage (for 10 min) has been found to be accompanied by an increase of the relative content of TH-positive neurons in some CNS areas of both mollusc species. In intact Chlamys farreri nipponensis, the presence of NOS in the CNS and its significant activation under temperature stress might have possibly been an important neuroprotective component of stress reaction in some mollusc species.     

THE NEURAL ARCHITECTURE OF THE SEA ANEMONE MIMETR1DIUM CRYPTUM

Mimetridium cryptum is a slender, elongated, New Zealand seaanemone. It shows fast contractions in the retractor musclesof its mesenteries, oral disc radial muscles, and tentacle longitudinalmuscles. The nervous system shows considerable regional differentiationin orientation of neurons, range of diameters of nerve fibers,and density of nerve net. Fast-contracting muscles are overlainby relatively dense nerve net, with many nerve fibers of morethan 2 ,µ diameter; slow-contracting muscles are overlainby a sparse nerve net whose nerve fibers are about 1µin diameter. A tendency for nerve fibers to run parallel ismarked in some regions. Individual neurons may run from onestructure to another, and even pass from the endoderm of themesenteries to the ectoderm of the oral disc.     

The giant fiber system in the forelegs (whips) of the whip spider Heterophrynus elaphus Pocock (Arachnida: Amblypygi)

Summary The front legs of the whip spider H. elaphus are strongly modified to serve sensory functions. They contain several afferent nerve fibers which are so large that their action potentials can be recorded externally through the cuticle. In recordings from the tarsus 7 different types of afferent spikes were identified; 6 additional types of afferent spikes were discriminated in recordings from the tibia and femur. Most of the recorded potentials could be attributed to identifiable neurons serving different functions. These neurons include giant interneurons and giant fibers from diverse mechanoreceptors such as slit sense organs, trichobothria, and a joint receptor. In the present report these neurons are characterized using electrophysiological and histological methods. Their functions are discussed in the context of the animal's behavior.Abbreviations GN giant neuron - S segment     

Localization of substance P, calcitonin gene related peptide and galanin in the nerve fibers of porcine cystic ovaries

In a previous study, we showed that both the noradrenergic and cholinergic component of ovarian innervation is markedly changed in porcine cystic ovaries. The present study is aimed at elucidating the distribution pattern of substance P- (SP), calcitonin gene related peptide CGRP- and/or galanin (GAL)-containing nerve fibers within porcine cystic ovaries. The status polycysticus was induced by dexamethasone phosphate disodium salt i.m. injections performed from the 7(th) until the 21(st) day of the first studied estrous cycle. During the same period of time, gilts of the control group received saline. All animals were slaughtered on the expected 11(th) day of the second studied estrous cycle, and their ovaries were collected. When compared to control gonad, a distinct difference in the distribution pattern and the density of SP-, CGRP- and/or GAL-immunoreactive (GAL-IR) nerve fibers was observed. Thus, unlike in the control gonad, SP- and/or CGRP-IR perivascular nerve fibers were found to supply medullar blood vessels of polycystic ovary. Furthermore, the number of GAL-IR nerve fibers contributing to the ground plexus in polycystic ovaries was higher than that observed in the control gonads. Thus, as may be judged from the profound changes in the distribution pattern of differently chemically coded afferent terminals within polycystic gonads, it appears possible that neuropeptides released from these terminals may take part in the etiopathogenesis of this disorder.     

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.