Longitudinal Neurite Bundles Research Articles

The nervous system in the cyclostome bryozoan Crisia eburnea as revealed by transmission electron and confocal laser scanning microscopy

IntroductionAmong bryozoans, cyclostome anatomy is the least studied by modern methods. New data on the nervous system fill the gap in our knowledge and make morphological analysis much more fruitful to resolve some questions of bryozoan evolution and phylogeny.ResultsThe nervous system of cyclostome Crisia eburnea was studied by transmission electron microscopy and confocal laser scanning microscopy. The cerebral ganglion has an upper concavity and a small inner cavity filled with cilia and microvilli, thus exhibiting features of neuroepithelium. The cerebral ganglion is associated with the circumoral nerve ring, the circumpharyngeal nerve ring, and the outer nerve ring. Each tentacle has six longitudinal neurite bundles. The body wall is innervated by thick paired longitudinal nerves. Circular nerves are associated with atrial sphincter. A membranous sac, cardia, and caecum all have nervous plexus.ConclusionThe nervous system of the cyclostome C. eburnea combines phylactolaemate and gymnolaemate features. Innervation of tentacles by six neurite bundles is similar of that in Phylactolaemata. The presence of circumpharyngeal nerve ring and outer nerve ring is characteristic of both, Cyclostomata and Gymnolaemata. The structure of the cerebral ganglion may be regarded as a result of transformation of hypothetical ancestral neuroepithelium. Primitive cerebral ganglion and combination of nerve plexus and cords in the nervous system of C. eburnea allows to suggest that the nerve system topography of C. eburnea may represent an ancestral state of nervous system organization in Bryozoa. Several scenarios describing evolution of the cerebral ganglion in different bryozoan groups are proposed.

Nervous system development in the Pacific oyster, Crassostrea gigas (Mollusca: Bivalvia)

BackgroundBivalves comprise a large, highly diverse taxon of invertebrate species. Developmental studies of neurogenesis among species of Bivalvia are limited. Due to a lack of neurogenesis information, it is difficult to infer a ground pattern for Bivalvia. To provide more comprehensive morphogenetic data on bivalve molluscs and relationships among molluscan clades, we investigated neurogenesis in the Pacific oyster, Crassostrea gigas, from the appearance of the first sensory cells to the formation of the larval ganglionic nervous system by co-immunocytochemistry of the neuronal markers FMRFamide or 5-HT and vesicular acetylcholine transporter (VAChT).ResultsNeurogenesis begins with the emergence of the apical serotonin-immunoreactive (5-HT-ir) sensory cells and paired sensory posttrochal dorsal and ventral FMRFamide-immunoreactive (FMRFamide-ir) cells at the early trochophore stage. Later, at the early veliger stage, the apical organ (AO) includes 5-HT-ir, FMRFamide-ir, and VAChT-ir cells. At the same stage, VAChT-ir cells appear in the posterior region of larvae and send axons towards the AO. Thus, FMRFamide-ir neurites and VAChT-ir processes form scaffolds for longitudinal neurite bundles develop into the paired ventral nerve cords (VNC). Later-appearing axons from the AO/CG neurons join the neurite bundles comprising the VNC. All larval ganglia appear along the VNC as paired or fused (epiathroid) clusters in late veliger and pediveliger larvae. We observed the transformation of the AO into the cerebral ganglia, which abundantly innervated the velum, and the transformation of ventral neurons into the pedal ganglia, innervating the foot, gills, and anterior adductor muscle. The visceral ganglia appear last in the pediveliger oyster and innervate the visceral mass and posterior adductor of premetamorphic larvae. In addition, a local FMRFamide-ir network was detected in the digestive system of pediveliger larvae. We identified VAChT-ir nervous elements in oyster larvae, which have not been observed previously in molluscs. Finally, we performed a morphology-based comparative analysis of neuronal structures among bivalve, conchiferan, and aculiferan species.ConclusionsWe described the development of the nervous system during the larval development in Crassostrea gigas. These data greatly advance the currently limited understanding of neurodevelopment in bivalves and mollusks, which has hampered the generation of a ground pattern reconstruction of the last common ancestor of Mollusca. Our morphological data support phylogenomic data indicating a closer Bivalvia-Gastropoda sister group relationship than the Bivalvia-Scaphopoda (Diasoma) group relationship.

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.

Neuroanatomy of Hyalinella punctata: Common patterns and new characters in phylactolaemate bryozoans.

Studies on the bryozoan adult nervous system employing immunocytochemical techniques and confocal laser scanning microscopy are scarce. To gain a better view into the structure and evolution of the nervous system of the Phylactolaemata, the earliest extant branch and sister taxon to the remaining Bryozoa, this work aims to characterize the nervous system of Hyalinella punctata with immunocytochemical techniques and confocal laser scanning microscopy. The cerebral ganglion is located between the anus and the pharynx and contains a lumen. Two ganglionic horns and a circum-oral nerve ring emanate from the cerebral ganglion. The pharynx is innervated by a diffuse neural plexus with two prominent neurite bundles. The caecum is innervated by longitudinal neurite bundles and a peripheral plexus. The intestine is characterized by longitudinal and circular neurite bundles, mostly near the anus. Novel putative sensory cells were found in the foregut and intestine. The tentacle sheath is innervated by a diffuse neural plexus, which emanates from several neurite bundles from the cerebral ganglion, but also parts of the pharyngeal plexus. There are six tentacle neurite bundles of intertentacular origin. The retractor muscles are innervated by two thin neurite bundles. Several characters are described herein for the first time in Phylactolaemata: Longitudinal neurite bundles and a peripheral plexus of the caecum, putative sensory structures of the gut, retractor muscle innervation, specific duplicature band neurite bundles. The tentacle innervation differs from previous descriptions of phylactolaemates regarding the origin of the three abfrontal neurite bundles. In general, most organ systems are innervated by a diffuse plexus in phylactolaemates as opposed to gymnolaemates. In contrast to the Gymnolaemata, representatives of Phylactolaemata show a higher number of tentacle nerves. Although the plesiomorphic condition for zooidal features among bryozoans remains unclear, having a diffuse nerve plexus may represent an ancestral feature for freshwater bryozoans.

The nervous system of the lophophore in the ctenostome Amathia gracilis provides insight into the morphology of ancestral ectoprocts and the monophyly of the lophophorates.

BackgroundThe Bryozoa (=Ectoprocta) is a large group of bilaterians that exhibit great variability in the innervation of tentacles and in the organization of the cerebral ganglion. Investigations of bryozoans from different groups may contribute to the reconstruction of the bryozoan nervous system bauplan. A detailed investigation of the polypide nervous system of the ctenostome bryozoan Amathia gracilis is reported here.ResultsThe cerebral ganglion displays prominent zonality and has at least three zones: proximal, central, and distal. The proximal zone is the most developed and contains two large perikarya giving rise to the tentacle sheath nerves. The neuroepithelial organization of the cerebral ganglion is revealed. The tiny lumen of the cerebral ganglion is represented by narrow spaces between the apical projections of the perikarya of the central zone. The cerebral ganglion gives rise to five groups of main neurite bundles of the lophophore and the tentacle sheath: the circum-oral nerve ring, the lophophoral dorso-lateral nerves, the pharyngeal and visceral neurite bundles, the outer nerve ring, and the tentacle sheath nerves. Serotonin-like immunoreactive nerve system of polypide includes eight large perikarya located between tentacles bases. There are two analmost and six oralmost perikarya with prominent serotonergic “gap” between them. Based on the characteristics of their innervations, the tentacles can be subdivided into two groups: four that are near the anus and six that are near the mouth. Two longitudinal neurite bundles - medio-frontal and abfrontal - extend along each tentacle.ConclusionThe zonality of the cerebral ganglion, the presence of three commissures, and location of the main nerves emanating from each zone might have caused by directive innervation of the various parts of the body: the tentacles sheath, the lophohpore, and the digestive tract. Two alternative scenarios of bryozoan lophophore evolution are discussed. The arrangement of large serotonin-like immunoreactive perikarya differs from the pattern previously described in ctenostome bryozoans. In accordance with its position relative to the same organs (tentacles, anus, and mouth), the lophophore outer nerve ring corresponds to the brachiopod lower brachial nerve and to the phoronid tentacular nerve ring. The presence of the outer nerve ring makes the lophophore innervation within the group (clade) of lophophorates similar and provides additional morphological evidence of the lophophore homology and monophyly of the lophophorates.Electronic supplementary materialThe online version of this article (doi:10.1186/s12862-016-0744-7) contains supplementary material, which is available to authorized users.

Nervous system development in the fairy shrimp Branchinella sp. (Crustacea: Branchiopoda: Anostraca): Insights into the development and evolution of the branchiopod brain and its sensory organs.

Using immunohistochemical labeling against acetylated a-tubulin and serotonin in combination with confocal laser scanning microscopy and 3D-reconstruction, we investigated the temporary freshwater pond inhabitant Branchinella sp. (Crustacea: Branchiopoda: Anostraca) for the first time to provide detailed data on the development of the anostracan nervous system. Protocerebral sense organs such as the nauplius eye and frontal filament organs are present as early as the hatching stage L0. In the postnaupliar region, two terminal pioneer neurons grow from posterior to anterior to connect the mandibular neuromeres. The first protocerebral neuropil to emerge is not part of the central complex but represents the median neuropil, and begins to develop from L0+ onwards. In stage L3, the first evidence of developing compound eyes is visible. This is followed by the formation of the visual neuropils and the neuropils of the central complex in the protocerebrum. From the deutocerebral lobes, the projecting neuron tract proceeds to both sides of the lateral protocerebrum, forming a chiasma just behind the central body. In the postnaupliar region, the peripheral nervous system, commissures and connectives develop along an anterior-posterior gradient after the fasciculation of the terminal pioneer neurons with the mandibular neuromere. The peripheral nervous system in the thoracic segments consists of two longitudinal neurite bundles on each side which connect the intersegmental nerves, together with the ventral nervous system forming an orthogon-like network. Here, we discuss, among other things, the evidence of a fourth nauplius eye nerve and decussating projecting neuron tract found in Branchinella sp., and provide arguments to support our view that the crustacean frontal filament (organ) and onychophoran primary antenna are homologous. J. Morphol. 277:1423-1446, 2016. © 2016 Wiley Periodicals, Inc.

Development of the nervous system in Solenogastres (Mollusca) reveals putative ancestral spiralian features.

BackgroundThe Solenogastres (or Neomeniomorpha) are a taxon of aplacophoran molluscs with contentious phylogenetic placement. Since available developmental data on non-conchiferan (that is, aculiferan) molluscs mainly stem from polyplacophorans, data on aplacophorans are needed to clarify evolutionary questions concerning the morphological features of the last common ancestor (LCA) of the Aculifera and the entire Mollusca. We therefore investigated the development of the nervous system in two solenogasters, Wirenia argentea and Gymnomenia pellucida, using immunocytochemistry and electron microscopy.ResultsNervous system formation starts simultaneously from the apical and abapical pole of the larva with the development of a few cells of the apical organ and a posterior neurogenic domain. A pair of neurite bundles grows out from both the neuropil of the apical organ and the posterior neurogenic domain. After their fusion in the region of the prototroch, which is innervated by an underlying serotonin-like immunoreactive (−LIR) plexus, the larva exhibits two longitudinal neurite bundles - the future lateral nerve cords. The apical organ in its fully developed state exhibits approximately 8 to 10 flask-shaped cells but no peripheral cells. The entire ventral nervous system, which includes a pair of longitudinal neurite bundles (the future ventral nerve cords) and a serotonin-LIR ventromedian nerve plexus, appears simultaneously and is established after the lateral nervous system. During metamorphosis the apical organ and the prototrochal nerve plexus are lost.ConclusionsThe development of the nervous system in early solenogaster larvae shows striking similarities to other spiralians, especially polychaetes, in exhibiting an apical organ with flask-shaped cells, a single pair of longitudinal neurite bundles, a serotonin-LIR innervation of the prototroch, and formation of these structures from an anterior and a posterior neurogenic domain. This provides evidence for an ancestral spiralian pattern of early nervous system development and a LCA of the Spiralia with a single pair of nerve cords. In later nervous system development, however, the annelids deviate from all other spiralians including solenogasters in forming a posterior growth zone, which initiates teloblastic growth. Since this mode of organogenesis is confined to annelids, we conclude that the LCA of both molluscs and spiralians was unsegmented.

Immunohistochemical and ultrastructural analysis of the muscular and nervous systems in the interstitial polychaete Polygordius appendiculatus (Annelida)

The systematic position of Polygordiidae is still under debate. They have been assigned to various positions among the polychaetes. Recent molecular analyses indicate that they might well be part of a basal radiation in Annelida, suggesting that certain morphological characters could represent primitive character traits adopted from the annelid stem species. To test this hypothesis, an investigation of the muscular and nervous systems by means of immunological staining and confocal laser scanning microscopy and transmission electron microscopy was conducted. With the exception of the brain, the nervous system is entirely basiepidermal and consists of the brain, the esophageal connectives, the subesophageal region, the ventral nerve cord and several smaller longitudinal nerves. These are connected by a considerable number of ring nerves in each segment. The ventral nerve cord is made up of closely apposed longitudinal neurite bundles, a median and two larger lateral ones. Since distinct ganglia are lacking, it represents a medullary cord. The muscular system mainly consists of longitudinal fibers, regularly distributed oblique muscles and strong septa. The longitudinal fibers form a right and a left unit separated along the dorsal midline, each divided into a dorsal and ventral part by the oblique muscles. Anteriorly, the longitudinal musculature passes the brain and terminates in the prostomium. There is no musculature in the palps. In contrast to earlier observations, regularly arranged minute circular muscle fibers are present. Very likely, a basiepithelial and non-ganglionic organization of the ventral nerve cord as well as an orthogonal nervous system represent plesiomorphic characters. The same applies for the predominance of longitudinal muscle fibers.

The nervous system of Isodiametra pulchra (Acoela) with a discussion on the neuroanatomy of the Xenacoelomorpha and its evolutionary implications

IntroductionAcoels are microscopic marine worms that have become the focus of renewed debate and research due to their placement at the base of the Bilateria by molecular phylogenies. To date, Isodiametra pulchra is the most promising “model acoel” as it can be cultured and gene knockdown can be performed with double-stranded RNA. Despite its well-known morphology data on the nervous system are scarce. Therefore we examined this organ using various microscopic techniques, including histology, conventional histochemistry, electron microscopy, and immunocytochemistry in combination with CLSM and discuss our results in light of recently established phylogenies.ResultsThe nervous system of Isodiametra pulchra consists of a bilobed brain with a dorsal posterior commissure, a frontal ring and tracts, four pairs of longitudinal neurite bundles, as well as a supramuscular and submuscular plexus. Serotonin-like immunoreactivity (SLI) is displayed in parts of the brain, the longitudinal neurite bundles and a large part of the supramuscular plexus, while FMRFamide-like immunoreactivity (RFLI) is displayed in parts of the brain and a distinct set of neurons, the longitudinal neurite bundles and the submuscular plexus. Despite this overlap SLI and RFLI are never colocalized. Most remarkable though is the presence of a distinct functional neuro-muscular system consisting of the statocyst, tracts, motor neurons and inner muscles, as well as the presence of various muscles that differ with regard to their ultrastructure and innervation.ConclusionsThe nervous system of Isodiametra pulchra consists of an insunk, bilobed brain, a peripheral part for perception and innervation of the smooth body-wall musculature as well as tracts and motor neurons that together with pseudostriated inner muscles are responsible for steering and quick movements. The insunk, bilobed brains with two to three commissures found in numerous acoels are homologous and evolved from a ring-commissural brain that was present in the stem species of acoelomorphs. The acoelomorph brain is bipartite, consisting of a Six3/6-dependend animal pole nervous system that persists throughout adulthood and an axial nervous system that does not develop by exhibiting a staggered pattern of conserved regulatory genes as in other bilaterians but by a nested pattern of these genes. This indicates that acoelomorphs stem from an ancestor with a simple brain or with a biphasic life cycle.

The nervous system of Neodasys chaetonotoideus (Gastrotricha: Neodasys) revealed by combining confocal laserscanning and transmission electron microscopy: evolutionary comparison of neuroanatomy within the Gastrotricha and basal Protostomia

We present a reconstruction of the nervous system of Neodasys chaetonotoideus Remane, 1927 (Gastrotricha, Chaetonotida) based on different microscopical methods: (1) immunohistochemistry (anti-acetylated α- and β-tubulin-, anti-5-HT- and anti-FMRFamide labelling) and (2) histochemistry (labelling of musculature and nuclei) by the means of confocal laser scanning microscopy (cLSM) and (iii) ultrastructure by means of transmission electron microscopy (TEM). All parts of the nervous system contain structures with an immunoreaction against the used immunohistochemical markers and labelling of histochemical markers. Results of both techniques (cLSM, TEM) reveal that the nervous system of N. chaetonotoideus is composed of a “dumb-bell-shaped” brain and one pair of posterior longitudinal neurite bundles. The brain is made up of a pair of laterally located clusters of neuronal somata, a large dorsal interconnecting dorsal commissure and two tiny ventral commissures in the region of the lateral clusters. From this, it follows that the brain is circumpharyngeal in position. The innervation of the head region is conducted by three pairs of anterior-directed neurite bundles. We describe here the gross anatomy of the nervous system and give additional details of the ultrastructure and the 5-HT and RFamide-like IR components of the nervous system. We compare our newly obtained data with already published data on the nervous system of gastrotrichs to reconstruct the hypothetical ground pattern of the nervous system in Gastrotricha, respectively, in Macrodasyida.

Acetylcholinesterase activity in the developing and regenerating nervous system of the acoel Symsagittifera roscoffensis

AbstractBery,A.andMarti´nez, P. 2010. Acetylcholinesterase activity in the developingand regenerating nervous system of the acoel Symsagittifera roscoffensis. —ActaZoologica (Stockholm) 00:1–10.The use of the cholinergic system is wid espread in the animal kingdom. It con-trols different processes, including reproduction and neural transmission. How-ever, its evolutionary history is not yet well understood. For instance, the roleplayed by the cholinergic system in the nervous system of basal bilaterian taxa,where the first signs of architectural complexity appear, is still unknown. Here,we describe the structure of the cholinergic system during the development andregeneration of the acoel flatworm Symsagittifera roscoffensis, using acetylcholines-terase (AchE) activity as a marker. In this species, AchE activity is observed at alldevelopmental stages, including in the early embryos. The juvenile and adultpatterns reveal the presence of a comple x nervous system that includes threepairs of longitudinal neurite bundles, whi ch are connected to an anterior central-ized mass of neurons and neural processes formed by two pairs of connectivesand four commissures. The power of the technique also allows the detection ofnewly born neurons as they are incorpo rated into the growing nervous system(during regeneration).Pedro Marti´nez, Departament de Gene`tica, Universitat de Barcelona, Av. Diag-onal, 645. E- 08028-Barcelona. Spain. E-mail: Pedro.martinez@ub.eduIntroductionAcetylcholine (Ach) is a molecule that has been detected inbacteria, fungi, plants and animals (Horiuchi et al. 2003).The synthesis of the neurotransmitter Ach is modulated bythe enzymes choline acetyl transferase and carnitine acetyltransferase. Conversely, the determination of neurotransmis-sion at cholinergic synapses is produced by the hydrolysis ofthis neurotransmitter mediated by the enzyme acetylcholines-terase (AchE) (Silman and Sussman 2008). The biologicalfunction of Ach has been studied extensively in bilaterians,especially insects and vertebrates (Le Nove`re et al. 2002).However, in organisms without a nervous system and in morebasal metazoans, the physiological function of Ach has yet tobe determined, although it may be a local mediator.Moreover, even in biological systems that have a well-orga-nized nervous system, the presence of components of the cho-linergic system is not restricted to neurons (Ellwanger andNickel 2006; Denker et al. 2008; Wessler and Kirkpatrick2008).To understand the origin and the diversification of the neu-ral cholinergic system, we studied AchE activity in a memberof the supposedly earliest offshoot of the Bilateria, the phylumAcoela. Molecular phylogenies indicate that the Acoela is thesister group of the remaining Bilateria (Ruiz-Trillo et al. 1999,2004; Halanych 2004; Sempere et al. 2007; Wallberg et al.2007; Hejnol et al. 2009). According to Wallberg et al. (2007)Acoela and Nemertodermatida are two early bilaterian clades.However, a recent phylogenomic analysis suggests, on thecontrary, the monophyletic nature of the Acoelomorpha(Hejnol et al. 2009). Obviously, this is still an unsettled issue.Nevertheless, Acoela probably represents the first group witha ‘centralized’ nervous system (Tyler and Hyra 1998; Reuteret al. 2001) or, more specifically, with a mass of neurons

Steps towards a centralized nervous system in basal bilaterians: Insights from neurogenesis of the acoel Symsagittifera roscoffensis

Due to its proposed basal position in the bilaterian Tree of Life, Acoela may hold the key to our understanding of the evolution of a number of bodyplan features including the central nervous system. In order to contribute novel data to this discussion we investigated the distribution of α-tubulin and the neurotransmitters serotonin and RFamide in juveniles and adults of the sagittiferid Symsagittifera roscoffensis. In addition, we present the expression pattern of the neuropatterning gene SoxB1. Adults and juveniles exhibit six serotonergic longitudinal neurite bundles and an anterior concentration of serotonergic sensory cells. While juveniles show an "orthogon-like" arrangement of longitudinal neurite bundles along the anterior-posterior axis, it appears more diffuse in the posterior region of adults. Commissures between the six neurite bundles are present only in the anterior body region of adults, while irregularly distributed individual neurites, often interconnected by serotonergic nerve cells, are found in the posterior region. Anti-RFamide staining shows numerous individual neurites around the statocyst. The orthogon-like nervous system of S.roscoffensis is confirmed by α-tubulin immunoreactivity. In the region of highest neurotransmitter density (i.e., anterior), the HMG-box gene SrSoxB1, a transcription factor known to be involved in neurogenesis in other bilaterians, is expressed in juvenile specimens. Accordingly, SoxB1 expression in S.roscoffensis follows the typical pattern of higher bilaterians that have a brain. Thus, our data support the notion that Urbilateria already had the genetic toolkit required to form brain-like neural structures, but that its morphological degree of neural concentration was still low.

Structure of the nervous system in Tubiluchus troglodytes (Priapulida)

Abstract. The nervous system of the meiobenthic priapulid species Tubiluchus troglodytes is described by immunohistochemistry and confocal laser scanning microscopy. The brain is circumpharyngeal, consisting of a central ring of neuropil and both anterior and posterior somata. From the brain emerges a ventral nerve cord, which shows ganglion‐like swellings in the neck and caudal region. The introvert includes longitudinal neurite bundles running below and between the rows of scalids, with a small cluster of sensory cells under each scalid. In the body wall of the neck and trunk region, longitudinal and circular neurite bundles are present in an orthogonal pattern. The tail is innervated from the caudal swelling of the ventral nerve cord; it also includes longitudinal and circular bundles in an orthogonal pattern. The pharynx has a reticulated system of neurite bundles running between the pharyngeal teeth and fimbrillae. Below each tooth and fimbrilus is a ganglion‐like cluster of somata. The intestine is surrounded by a nerve net. The data on the nervous system are compared within other priapulids and with other species of Scalidophora (Kinorhyncha and Loricifera).