The nervous system of Tricladida. I. Neuroanatomy ofProcerodes littoralis (Maricola,Procerodidae): An immunocytochemical study

The organization of the nervous system ofProcerodes littoralis (Tricladida, Maricola, Procerodidae) was studied by immunocytochemistry, using antibodies to authentic flatworm neuropeptide F (NPF) (Moniezia expansa). Compared to earlier investigations of the neuroanatomy of tricladid flatworms, the pattern of NPF immunoreactivity inProcerodes littoralis reveals differences in the following respects: 1. Shape and structure of the brain. 2. Number and composition of longitudinal nerve cords. 3. Shape of branches of, and transverse connections between, main ventral nerve cords. 4. Composition of the pharyngeal nervous system. The rich innervation by NPF immunoreactive (IR) fibres and cells of the subepithelial muscle layer, the pharynx musculature and the musculature of the male copulatory apparatus indicates a neurotransmitter or neuromodulatory influence on muscular activity.     

The nervous system of Tricladida. IV. Neuroanatomy of Planaria torva (Paludicola,Planaridae): an immunocytochemical study

Characters of the nervous system have scarcely been used in consideration of the phylogenetic relationships of the Tricladida. In order to contribute characters toward this end, we studied the nervous system of Planaria torva with immunocytochemical methods and we compare the results with what is known of nervous system structure in other triclads. Antisera against four neuroactive substances were used: neuropeptide F (NPF), RFamide, GYIRFamide, and serotonin (5-HT). All four substances showed positive immunoreactions throughout the nervous system; and the patterns of distribution of the first three were similar, while 5-HT showed a distinct distribution. The organization of these nervous system elements in Planariidae and Dendrocoelidae resembles that of the Maricola but differs from that of the Dugesiidae. The results support the hypotheses that the Paludicola is a more advanced taxon than the Maricola and that the Planariidae and Dendrocoelidae together are a sister group of the Dugesiidae. This revised version was published online in July 2006 with corrections to the Cover Date.     

The nervous system of Tricladida. III. Neuroanatomy ofDendrocoelum lacteum andPolycelis tenuis (Plathelminthes,Paludicola): an immunocytochemical study

The organization of the nervous system ofDendrocoelum lacteum (Tricladida, Paludicola, Dendrocelidae) andPolycelis tenuis (Tricladida, Paludicola, Planariidae) was studied by immunocytochemical double staining, using neuropeptide RFamide and serotonin (5-HT) antisera on cryosections. The study confirmed the status of the main nerve cords (MCs) as the most important and stable of the longitudinal cords and supported the hypothesis of a common phylogenetic origin of the MCs in flatworms. The ganglion-like structures along the MCs at the beginning of transverse commissures and laterla branches showed a close contact with ventral fibres of the submuscular nerve plexus indicating an origin from crossing points of insunken ring commissures. The distributional pattern and morphology of the RFamide-IR cell bodies inD. lacteum corresponded to that of neurosecretory cells. Most RFamide-IR cells were unipolar and rounded while 5-HT-IR cells were uni- bi- and multipolar. The neutropile consisted of a dense RFamide-IR and a loose 5-HT-IR network. RFamide dominated in all parts of the genital plexus.     

Cholinergic, serotoninergic and peptidergic components of the nervous system of Discocotyle sagittata (Monogenea: Polyopisthocotylea)

Cholinergic, serotoninergic (5-HT) and peptidergic neuronal pathways have been demonstrated in both central and peripheral nervous systems of adult Discocotyle sagittata, using enzyme histochemistry and indirect immunocytochemistry in conjunction with confocal scanning laser microscopy. Antisera to 2 native flatworm neuropeptides, neuropeptide F and the FMRFamide-related peptide (FaRP), GNFFRFamide, were employed to detect peptide immunoreactivity. The CNS is composed of paired cerebral ganglia and connecting dorsal commissure, together with several paired longitudinal nerve cords. The main longitudinal nerve cords (Iateral, ventral and dorsal) are interconnected at intervals by a series of annular cross-connectives, producing a ladder-like arrangement typical of the platyhelminth nervous system. At the level of the haptor, the ventral cords provide nerve roots which innervate each of the 8 clamps. Cholinergic and peptidergic neuronal organisation was similar, but distinct from that of the serotoninergic components. The PNS and reproductive system are predominantly innervated by peptidergic neurones.     

Immunoreactive pancreatic polypeptide (PP) occurs in the central and peripheral nervous system: Preliminary immunocytochemical observations

Summary Pancreatic polypeptide (PP) is a candidate hormone of unknown physiological significance. It is produced by a population of endocrine cells in the pancreas. In the present study a PP-like peptide was found to occur in the mammalian and avian central and peripheral nervous systems. Immunoreactive nerve fibres and nerve cell bodies were widely distributed in the brain. Dense accumulations of nerve fibres occurred in the following areas: nucleus accumbens, interstitial nucleus of the stria terminalis, para- and periventricular hypothalamic nuclei, and medial preoptic area. In addition, nerve fibres were regularly seen in cortical areas. Immunoreactive perikarya were observed in the following regions: cortex, nucleus accumbens, neostriatum and septum. In the gut, immunoreactive nerve fibers were distributed in the myenteric plexus, in smooth muscle, around blood vessels, and in the core of the villi. Immunoreactive perikarya occurred in the submucosal and myenteric plexus, suggesting that PP immunoreactive nerves are intrinsic to the gut.In the species examined, the neuronal PP-like peptide could be demonstrated with an antiserum raised against avian PP, but not with those raised against bovine or human PP. Thus, neuronal PP is distinct from the PP that occurs in pancreatic endocrine cells.     

Crustacean cardioactive peptide in the nervous system of the locust,Locusta migratoria: an immunocytochemical study on the ventral nerve cord and peripheral innervation

Summary Crustacean cardioactive peptide-immunoreactive neurons occur in the entire central nervous system of Locusta migratoria. The present paper focuses on mapping studies in the ventral nerve cord and on peripheral projection sites. Two types of contralaterally projecting neurons occur in all neuromers from the subesophageal to the seventh abdominal ganglia. One type forms terminals at the surface of the thoracic nerves 6 and 1, the distal perisympathetic organs, the lateral heart nerves, and on ventral and dorsal diaphragm muscles. Two large neurons in the anterior part and several neurons of a different type in the posterior part of the terminal ganglion project into the last tergal nerves. In the abdominal neuromers 1–7, two types of ipsilaterally projecting neurons occur, one of which gives rise to neurosecretory terminals in the distal perisympathetic organs, in peripheral areas of the transverse, stigmata and lateral heart nerves. Four subesophageal neurons have putative terminals in the neurilemma of the nervus corporis allati II, and in the corpora allata and cardiaca. In addition, several immunoreactive putative interneurons and other neurons were mapped in the ventral nerve cord. A new in situ whole-mount technique was essential for elucidation of the peripheral pathways and targets of the identified neurons, which suggest a role of the peptide in the control of heartbeat, abdominal ventilatory and visceral muscle activity.Abbreviations AG abdominal ganglia - AM alary muscle - AMN alary muscle nerve - CA corpus allatum - CC corpus cardiacum - dPSO distal perisympathetic organ - LHN lateral heart nerve - LT CCAP-immunoreactive lateral tract - NCA nervus corporis allati - NCC nervus corporis cardiaci - NM neuromer - PMN paramedian nerve - PSO perisympathetic organ - SOG subesophageal ganglion - VDM ventral diaphragm muscles - VNC ventral nerve cord     

SALMFamide2 and serotonin immunoreactivity in the nervous system of some acoels (Xenacoelomorpha)

Acoel worms are simple, often microscopic animals with direct development, a multiciliated epidermis, a statocyst, and a digestive parenchyma instead of a gut epithelium. Morphological characters of acoels have been notoriously difficult to interpret due to their relative scarcity. The nervous system is one of the most accessible and widely used comparative features in acoels, which have a so‐called commissural brain without capsule and several major longitudinal neurite bundles. Here, we use the selective binding properties of a neuropeptide antibody raised in echinoderms (SALMFamide2, or S2), and a commercial antibody against serotonin (5‐HT) to provide additional characters of the acoel nervous system. We have prepared whole‐mount immunofluorescent stainings of three acoel species: Symsagittifera psammophila (Convolutidae), Aphanostoma pisae, and the model acoel Isodiametra pulchra (both Isodiametridae). The commissural brain of all three acoels is delimited anteriorly by the ventral anterior commissure, and posteriorly by the dorsal posterior commissure. The dorsal anterior commissure is situated between the ventral anterior commissure and the dorsal posterior commissure, while the statocyst lies between dorsal anterior and dorsal posterior commissure. S2 and serotonin do not co‐localise, and they follow similar patterns to each other within an animal. In particular, S2, but not 5‐HT, stains a prominent commissure posterior to the main (dorsal) posterior commissure. We have for the first time observed a closed posterior loop of the main neurite bundles in S. psammophila for both the amidergic and the serotonergic nervous system. In I. pulchra, the lateral neurite bundles also form a posterior loop in our serotonergic nervous system stainings.     

Distribution,abundance, and size of rhabdoids in Dugesia polychroa. (Turbellaria: Tricladida)

The ontogenetic changes in the distribution, abundance, and size of rhabdoids were examined in an undisturbed laboratory population of Dugesia polychroa. Irrespective of triclad age, rhabdoids in the epidermis and parenchyma were more abundant on the dorsal than the ventral side of the body. No significant differences were found in the abundance of epidermal or parenchymal rhabdoids among the anterior, medial, and posterior regions of the body. Rhabdoid number and size changed significantly with triclad age, with a marked depression coinciding with the onset of cocoon production. Rhabdoid discharge was correlated with physical and/or physiological disturbance and occurred in the absence of any overt environmental disturbance. Simple allometric relationships were observed between rhabdoid size and number on one hand and body plan area on the other. Different allometric trends were observed from field-collected individuals compared to the undisturbed laboratory population. The potential function of rhabdoids in the Tricladida is discussed in light of these findings.     

The female gonad in two species of Microplana (Platyhelminthes,Tricladida, Rhynchodemidae): Ultrastructural and cytochemical investigations

The female gonad of the land planarians Microplana scharffi and Microplana terrestris consists of two small germaria located ventrally in the anterior third of the body and of two ventro‐lateral rows of oblong vitelline follicles distributed between the intestinal pouches. Both these structures are enveloped by a tunica composed of an outer extracellular lamina and an inner sheath of accessory cells. Oocyte maturation is characterized by the appearance of chromatoid bodies and the development of endoplasmic reticulum and Golgi complexes. These organelles appear to be correlated with the production of egg granules with a fenestrated/granular content of medium electron density, about 4–5 μm in diameter, which remain dispersed in the ooplasm of mature oocytes. On the basis of cytochemical tests showing their glycoprotein composition, and their localization in mature oocytes, these egg granules have been interpreted as yolk. In the vitelline follicles, vitellocytes show the typical features of secretory cells with well‐developed rough endoplasmic reticulum and Golgi complexes involved in the production of eggshell globules and yolk. The eggshell globules, which appear to arise from repeated coalescences of two types of Golgi‐derived vesicles, contain polyphenols and, when completely mature, they measure about 1–1,2 μm in diameter and show a meandering/concentric content pattern as is typical of the situation observed in most Proseriata and Tricladida. Mature vitellocytes also contain a large amount of glycogen and lipids as further reserve material. On the basis of the ultrastructural features of the female gonad and in relation to the current literature the two species of rhynchodemids investigated appear to be closely related to the freshwater planarians belonging to the family Dugesiidae. J. Morphol. 2009. © 2009 Wiley‐Liss, Inc.     

The formation of the nervous system during larval development in Triops cancriformis (Bosc) (crustacea,Branchiopoda): An immunohistochemical survey

We provide data of the development of thenervous system during the first five larval stages of Triops cancriformis. We use immunohistochemical labeling (against acetylated α‐tubulin, serotonin, histamine, and FMRFamide), confocal laser scanning microscopy analysis, and 3D‐reconstruction. The development of the nervous system corresponds with the general anamorphic development in T. cancriformis. In larval stage I (L I), all brain parts (proto‐, deuto‐, and tritocerebrum), the circumoral connectives, and the mandibular neuromere are already present. Also, the frontal filaments and the developing nauplius eye are already present. However, until stage L III, the nauplius eye only consists of three cups. Throughout larval development, the protocerebral network differentiates into distinct subdivisions. In the postnaupliar region, additional neuromeres and their commissures emerge in an anteroposterior gradient. The larval nervous system in L V consists of a differentiated protocerebrum including a central body, a nauplius eye comprising four cups, a circumoral nerve ring, mandibular‐ and postnaupliar neuromeres up to the seventh thoracic segment, each featuring an anterior and a posterior commissure, and two parallel connectives. The presence of a protocerebral bridge is questionable. The distribution of neurotransmitters in L I is restricted to the naupliar nervous system. Over the course of the five stages of development, neurotransmitter distribution also follows an anteroposterior gradient. Each neuromere is equipped with two ganglia innervating the locomotional appendages and possesses a specific neurotransmitter distribution pattern. We suggest a correlation between neurotransmitter expression and locomotion. J. Morphol., 2010. © 2010 Wiley‐Liss, Inc.     

Autoradiographic detection of indolamine and catecholamine neurons in the nervous system of Owenia fusiformis (Polychaeta,Annelida)

Summary Following immobilization stress the supraoptic nucleus exhibits an increased number of coarse, heavily immunostained fibers in the basal glia labyrinth. Ultrastructural immunocytochemistry demonstrates a labeling of the rough endoplasmic reticulum in the neurosecretory perikarya and granule-free, immunoreactive material in the axons adjacent to the basal glia labyrinth. Furthermore, a labeling of the intercellular clefts of the neuropil is demonstrable in the supraoptic nucleus. These results lead to the hypothesis that 1) vasopressin is synthesized and released in two forms, in a granule-bound form and in a granule-free, probably more soluble form, and that 2) the latter might be released already in the nuclear area into the intercellular clefts from where it may reach its target cells via the cerebrospinal fluid of the subarachnoid space.The excellent technical assistance of Mrs. Helga Prien is thankfully acknowledgedSupported by the Deutsche Forschungsgemeinschaft (Grant Nr. Kr 569/2) and the Stiftung Volkswagenwerk     

Architecture of the nervous system in mystacocarida (Arthropoda,crustacea)—An immunohistochemical study and 3D reconstruction

Mystacocarida is a species‐poor group of minute crustaceans with unclear phylogenetic affinities. Previous studies have highlighted the putative “primitiveness” of several mystacocarid features, including the architecture of the nervous system. Recent studies on arthropod neuroarchitecture have provided a wealth of characters valuable for phylogenetic reconstructions. To permit and facilitate comparison with these data, we used immunohistochemical labeling (against acetylated α‐tubulin, serotonin and FMRFamide) on the mystacocarid Derocheilocaris remanei, analyzing it with confocal laser‐scanning microscopy and 3D reconstruction. The mystacocarid brain is fairly elongated, exhibiting a complicated stereotypic arrangement of neurite bundles. However, none of the applied markers provided evidence of structured neuropils such as a central body or olfactory glomeruli. A completely fused subesophageal ganglion is not present, all segmental soma clusters of the respective neuromeres still being delimitable. The distinct mandibular commissure comprises neurite bundles from more anterior regions, leading us to propose that it may have fused with an ancestral posterior tritocerebral commissure. The postcephalic ventral nervous system displays a typical ladder‐like structure with separated ganglia which bears some resemblance to larval stages in other crustaceans. Ganglia and commissures are also present in the first three limbless “abdominal” segments, which casts doubt on the notion of a clear‐cut distinction between thorax and abdomen. An unpaired longitudinal median neurite bundle is present and discussed as a potential tetraconate autapomorphy. Additionally, a paired latero‐longitudinal neurite bundle extends along the trunk. It is connected to the intersegmental nerves and most likely fulfils neurohemal functions. We report the complete absence of serotonin‐ir neurons in the ventral nervous system, which is a unique condition in arthropods and herein interpreted as a derived character. J. Morphol., 2010. © 2009 Wiley‐Liss, Inc.     

Myoanatomy and serotonergic nervous system of plumatellid and fredericellid phylactolaemata (lophotrochozoa,ectoprocta)

The phylogenetic position of the Ectoprocta within the Lophotrochozoa is discussed controversially. For gaining more insight into ectoproct relationships and comparing it with other potentially related phyla, we analysed the myoanatomy and serotonergic nervous system of adult representatives of the Phylactolaemata (Plumatella emarginata, Plumatellavaihiriae, Plumatella fungosa, Fredericella sultana). The bodywall contains a mesh of circular and longitudinal muscles. On its distal end, the orifice possesses a prominent sphincter and continues into the vestibular wall, which has longitudinal and circular musculature. The tentacle sheath carries mostly longitudinal muscle fibres in Plumatella sp., whereas F. sultana also possesses regular circular muscle fibres. Three groups of muscles are associated with the lophophore: 1) Lophophoral arm muscles (missing in Fredericella), 2) epistome musculature and 3) tentacle musculature. The epistome flap is encompassed by smooth muscle fibres. A few fibres extend medially over the ganglion to its proximal floor. Abfrontal tentacle muscles have diagonally arranged muscle fibres in their proximal region, whereas the distal region is formed by a stack of muscles that resemble an inverted ‘V’. Frontal tentacle muscles show more variation and either possess one or two bases. The digestive tract possesses circular musculature which is striated except at the intestine where it is composed of smooth muscle fibres. The serotonergic nervous system is concentrated in the cerebral ganglion. From the latter a serotonergic nerve extends to each tentacle base. In Plumatella the inner row of tentacles at the lophophoral concavity lacks serotonergic nerves. Bodywall musculature is a common feature in many lophotrochozoan phyla, but among other filter feeders like the Ectoprocta is only present in the ‘lophophorate’ Phoronida. The longitudinal tentacle musculature is reminiscent of the condition found in phoronids and brachiopods, but differs to entoproct tentacles. Although this study shows some support for the ‘Lophophorata’, more comparative analyses of possibly related phyla are required. J. Morphol., 2011. © 2011 Wiley Periodicals, Inc.     

The larval nervous system of the penis worm Priapulus caudatus (Ecdysozoa)

The origin and extreme diversification of the animal nervous system is a central question in biology. While most of the attention has traditionally been paid to those lineages with highly elaborated nervous systems (e.g. arthropods, vertebrates, annelids), only the study of the vast animal diversity can deliver a comprehensive view of the evolutionary history of this organ system. In this regard, the phylogenetic position and apparently conservative molecular, morphological and embryological features of priapulid worms (Priapulida) place this animal lineage as a key to understanding the evolution of the Ecdysozoa (i.e. arthropods and nematodes). In this study, we characterize the nervous system of the hatching larva and first lorica larva of the priapulid worm Priapulus caudatus by immunolabelling against acetylated and tyrosinated tubulin, pCaMKII, serotonin and FMRFamide. Our results show that a circumoral brain and an unpaired ventral nerve with a caudal ganglion characterize the central nervous system of hatching embryos. After the first moult, the larva attains some adult features: a neck ganglion, an introvert plexus, and conspicuous secondary longitudinal neurites. Our study delivers a neuroanatomical framework for future embryological studies in priapulid worms, and helps illuminate the course of nervous system evolution in the Ecdysozoa.     

Central nervous system antigen (NS-5) and its presence during murine ontogenesis

An antiserum raised by immunization of C3H.SW/Sn mice with cerebellum from 4-day-old C57BL/6J mice recognizes a cell surface component(s) [NS-5] present in different degrees on various parts of the mouse central nervous system. When analyzed by an antiserum-and complement-mediated cell cytotoxicity test and by the ability of various tissues to absorb anti-NS-5 antiserum activity, the antigen(s) was detectable on cerebellum, retina, olfactory bulb, cortex, basal ganglia, and medulla, but not on nonneural tissues with the exception of mature spermatozoa and 4-day-old kidney. The antigen(s) detected by the anti-NS-5 antiserum was found in similar quantities on young and adult rat and mouse cerebellum; however, it was not detectable on any of 16 clonal cell lines derived from the rat central nervous system. During preimplantation stages of murine development, the antigen could be detected on all cells of (2–4)-cell and (8–16)-cell stages and on the trophoblastic cells of blastocysts by indirect immunoflourescence. Embryos on day 9 of gestation, the earliest stage tested after implantation, expressed the antigen(s), but expression was restricted to the nervous system.     

Structure and anterior regeneration of musculature and nervous system in Cirratulus cf. cirratus (Cirratulidae,Annelida)

Annelids provide suitable models for studying regeneration. By now, comprehensive information is restricted to only a few taxa. For many other annelids, comparative data are scarce or even missing. Here, we describe the regeneration of a member of the Cirratulus cirratus species complex. Using phalloidin‐labeling and antibody‐stainings combined with subsequent confocal laser scanning microscopy, we provide data about the organization of body wall musculature and nervous system of intact specimens, as well as about anteriorly regenerating specimens. Our analyses show that C. cf. cirratus exhibits a prominent longitudinal muscle layer forming a dorsal muscle plate, two ventral muscle strands and a ventral‐median muscle fiber. The circular musculature forms closed rings which are interrupted in the area of parapodia. The nervous system of C. cf. cirratus shows a typical rope‐ladder like arrangement and the circumesophageal connectives exhibit two separate roots leading to the brain. During regeneration, the nervous system redevelops remarkably earlier than the musculature, first constituting a tripartite loop‐like structure which later become the circumesophageal connectives. Regeneration of longitudinal musculature starts with diffuse ingrowth and subsequent structuring into the blastema. In contrast, circular musculature develops independently inside the blastema. Our findings constitute the first analysis of regeneration for a member of the Cirratuliformia on a structural level. Summarizing the regeneration process in C. cf. cirratus, five main phases can be subdivided: 1) wound closure, 2) blastema formation, 3) blastema differentiation, 4) resegmentation, and 5) growth, respectively elongation. Additionally, the described tripartite loop‐like structure of the regenerating nervous system has not been reported for any other annelid taxon. In contrast, the regeneration of circular and longitudinal musculature originating from different groups of cells seems to be a general pattern in annelid regeneration. J. Morphol. 275:1418–1430, 2014. © 2014 Wiley Periodicals, Inc.     

Glyoxylic acid induced fluorescence in the nervous system of Gyratrix hermaphroditus (Kalyptorhynchia,Polycystididae)

Catecholamines (CAs) are found in the neuropile of the brain, in 3 pairs of longitudinal nerve cords, in the transverse ventral commissure, in anterior ventral and dorsal nerves, in two pharyngeal nerve rings and in 24 neurons in the nervous system of Gyratrix hermaphroditus. The CA distribution pattern in compared with those of other neuroactive substances. Homology of neurons in the family of Polycystididae and in Plathelminthes in general is discussed.