Expression of synapsin and co-localization with serotonin and RFamide-like immunoreactivity in the nervous system of the chordoid larva of Symbion pandora (Cycliophora)

Abstract. Cycliophora is one of the most recently described metazoan phyla and hitherto includes only two species: Symbion pandora and Symbion americanus . With a very complex life cycle, cycliophorans are regarded as an enigmatic group with an uncertain phylogenetic position, although they are commonly considered lophotrochozoan protostomes. In order to extend the database concerning the distribution of immunoreactive substances in the free-swimming chordoid larva of S. pandora , we investigated synapsin immunoreactivity using fluorescence-coupled antibodies in combination with confocal laserscanning microscopy. Moreover, we analyzed the co-localization patterns of synapsin, serotonin, and RFamide-like immunoreactivity in the chordoid larva by 3D imaging technology based on the confocal microscopy image stacks. Synapsin is expressed in large parts of the bilobed anterior cerebral ganglion including anterior and dorsal projections. Two pairs of ventral neurites run longitudinally into the larval body of which the inner pair shows only weak, scattered synapsin immunoreactivity. In addition, a lateral synapsin immunoreactive projection emerges posteriorly from each ventral longitudinal axon. Double immunostaining shows co-localization of synapsin and serotonin in the cerebral ganglion, the outer and the inner ventral neurites, and the anterior projections. Synapsin and RFamide-like immunoreactivity co-occur in the cerebral ganglion, the outer ventral neurites, and the dorsal projections. Accordingly, the cerebral ganglion and the outer ventral neurites are the only neural structures that co-express the two neurotransmitters and synapsin. The overall neuroanatomical condition of the cycliophoran chordoid larva resembles much more the situation of adult rather than larval life cycle stages of a number of spiralian taxa.     

Three‐dimensional reconstruction of the musculature of various life cycle stages of the cycliophoran Symbion americanus

Cycliophora is a very recently described phylum of acoelomate metazoans with a complex life cycle and a phylogenetic position that has been under debate ever since its discovery in 1995. Symbion americanus, which lives attached to the mouthparts of the American lobster, Homarus americanus, represents the second species described for the phylum. Aiming to increase the morphological knowledge about this cryptic clade, the present study describes the muscle arrangement of the feeding stage, the attached Prometheus larva with the dwarf male inside, the free living male, the Pandora larva, and the chordoid larva of S. americanus using actin staining and confocal laser scanning microscopy. 3D reconstructions of the muscular systems are presented. In the feeding stage, circular muscles compose the buccal funnel aperture. In addition, a pair of muscles runs longitudinally in the buccal funnel. A complex sphincter was found just proximally to the anus, and six longitudinal muscles run from the trunk constriction (“neck”) in basal direction. The musculature of the larval stages and the dwarf male is very complex and includes longitudinal muscles that run dorsally and ventrally. In addition, we found dorso‐ventral muscles. The male has a complex posterior muscle apparatus in the vicinity of the penis. In this stage, X‐ and V‐shaped structures were identified on the dorsal and the ventral side, respectively. Pandora and chordoid larvae possess additional circular muscles. We discuss our findings with respect to muscle elements of other metazoan groups and the chordoid larva of Symbion pandora. J. Morphol., 2009. © 2008 Wiley‐Liss, Inc.     

Immunocytochemistry of the nervous system and the musculature of the chordoid larva of Symbion pandora (Cycliophora)

To date, the phylum Cycliophora comprises only one described extant species of acoelomate marine invertebrates, Symbion pandora. Adult specimens live commensally on the mouthparts of the Norwegian lobster, Nephrops norvegicus. Its complicated life cycle includes an asexually produced Pandora larva and a sexually produced chordoid larva. Despite detailed TEM investigations and its inclusion in recent molecular phylogenetic analyses, cycliophoran relationships still remain enigmatic. In order to increase the morphological database, I investigated the anatomy of the nervous system and the musculature of the chordoid larva by applying fluorescence-coupled antibodies against the neurotransmitters serotonin and FMRFamide, as well as FITC-coupled phalloidin to label filamentous F-actin, in combination with confocal laser scanning microscopy. The FMRFamidergic nervous system shows a bilobed anterior ganglion and one pair of ventral nerve cords, while serotonin is distributed in a scattered pattern in the anterior ganglion. In addition, there are two pairs of ventral serotonergic nerves, of which the inner pair fuses with the outer nerve cords in the posterior third of the larva. The musculature comprises an outer layer of six units of circular body wall muscles, several helicoid muscle fibers, a set of paired longitudinal muscles that span the entire anterior-posterior axis of the larva, and a few oblique muscle strands. Furthermore, an anterior muscle complex and one pair of posterior muscles are present. The chordoid organ consists of a number of distinct subunits that are each formed by a dense layer of circular muscle fibers.The overall arrangement of the oblique and longitudinal muscles as well as the body wall musculature in the chordoid larva of Symbion pandora exhibits similarities with the condition found in certain rotifers. This is congruent with some recent phylogenies based on 18S rRNA sequences but additional morphological, developmental, and molecular data are needed to clarify the phylogenetic relationships of Cycliophora.     

Comparative myoanatomy of cycliophoran life cycle stages

The metazoan phylum Cycliophora includes small cryptic epibionts that live attached to the mouthparts of clawed lobsters. The life cycle is complex, with alternating sexual and asexual generations, and involves several sessile and free‐living stages. So far, the morphological and genetic characterization of cycliophorans has been unable to clarify the phylogenetic position of the phylum. In this study, we add new details on the muscular anatomy of the feeding stage, the attached Prometheus larva, the dwarf male, and the female of one of the two hitherto described species, Symbion pandora. The musculature of the feeding stage is composed of myofibers that run longitudinally in the buccal funnel (two fibers) and in the trunk (variable number of fibers). The mouth opening is lined by a myoepithelial ring musculature. A complex myoepithelial sphincter is situated proximal to the anus. In the attached Prometheus larva, three longitudinal sets of myofilaments run dorsally, laterally, and ventrally along the entire anterior‐posterior body axis. The muscular architecture of the dwarf male is complex, especially close to the penis, in the posterior part of the body. An X‐shaped muscle structure is found on the dorsal side, whereas on the ventral side, longitudinal muscles and a V‐shaped muscle structure are present. These muscles are complemented by additional dorsoventral muscles. The mesodermal muscle fibers attach to the cuticle via the epidermis in all life cycle stages studied herein. The musculature of the female is similar to that of the Pandora larva of Symbion americanus and includes dorsoventral muscles and longitudinal muscles that run in the dorsal and ventral body region. Overall, our results reveal striking similarities in the muscular arrangement of the life cycle stages of both Symbion species. J. Morphol., 2010. © 2009 Wiley‐Liss, Inc.     

External morphology of the cycliophoran dwarf male: a comparative study of Symbion pandora and S. americanus

Cycliophora is a recently described phylum to which only two species have been assigned so far, Symbion pandora and S. americanus. The cycliophoran life cycle is complex and alternates between asexual and sexual stages. Although not recognized as an entirely independent free-swimming stage when the phylum was first described, the dwarf male has a remarkably complex bodyplan albeit its very small size (approx. 30–40 μm in length). Aiming to increase the knowledge on the gross morphology of the cycliophoran dwarf male, specimens from S. pandora and S. americanus were analyzed by scanning electron microscopy. In both species, anterior and ventral ciliated fields, as well as paired lateral sensorial organs, were identified, thus confirming previous observations. However, new details are described herein such as the penial pouch that encloses the penis. We compare our findings on both Symbion species with the data currently available on other metazoan dwarf males.     

First description of the serotonergic nervous system in a bdelloid rotifer: Macrotrachela quadricornifera Milne 1886 (Philodinidae)

Class Bdelloidea of phylum Rotifera comprises aquatic microinvertebrates that are known for both obligate parthenogenesis and for resisting desiccation through a dormant reversible state. In the frame of an investigation about the role of the nervous system in controlling life cycle, reproduction and dormancy, we describe the serotonergic system of a bdelloid, Macrotrachela quadricornifera, using serotonin immunohistochemistry and confocal laser scanning microscopy. Serotonin immunoreactivity is present in the cerebral ganglion, lateral nerve cords and peripheral neurites. The cerebral ganglion consists of perikarya that send neurites cephalically to the rostrum and corona. A pair of neurites exits the cerebral ganglion as lateral nerve cords, and proceeds caudally to the pedal ganglion where additional neurites enter the foot. Based on the location of serotonergic immunoreactivity, we hypothesize that the neurotransmitter is involved in both motor activity (e.g., ciliary beating, inchworm-like locomotion) and sensory activity. A comparison between the serotonergic nervous systems of M. quadricornifera and species of Monogononta reveals differences in the numbers and patterns of cerebral perikarya, peripheral perikarya, and periperhal neurites. These differences may have functional significance for understanding adaptations to specific environments and/or systematic significance for reconstructing the rotiferan ground pattern.     

A new type of loriciferan larva (Shira larva) from the deep sea of Shatsky Rise,Pacific Ocean

Loricifera is a phylum of minute animals that live exclusively in marine sediments. A total of 33 species have been described so far in this phylum; however, several more are already known from preliminary observations. Loriciferans are characterised by a complex life cycle, which involves a succession of several adult and larval stages. Here, we describe a new type of loriciferan larval stage: the Shira larva. The gross morphology of this larva is generally similar to that of the most prominent larval type of Loricifera, the so-called Higgins larva. However, the Shira larva possesses a number of unique features, namely (1) a single pair of anteroventral setae is present in the most anterior region of the abdomen, (2) the bases of the anteroventral setae are very large and swollen, (3) the thorax and abdomen are thinner than the introvert and (4) the abdominal region is divided into five sub-regions. Accordingly, we described the new species, Tenuiloricus shirayamai gen. nov. et sp. nov. (incertae sedis). The new findings are discussed from a comparative perspective with the Higgins larva as well as with the fossil of a putative loriciferan larval stage from the Middle Cambrian.     

The chordoid larva of Symbion pandora (Cycliophora) is a modified trochophore

Developmental and free-living stages of the chordoid larva of the cycliophoran species, Symbion pandora Funch and Kristensen 1995, were studied using light and electron microscopy. In the free-living stage of the larva, about 200 μm long, four ciliated areas are found: two anterior bands, a ventral ciliated field, and a posterior unit on the ventral side of the foot. The nervous system consists of a dorsal brain and a pair of ventral longitudinal nerves. A gut is absent. A pair of protonephridia, each with a single multiciliated terminal cell and at least one duct cell, is present. Nephridiopores are not localized. A pair of corsal ciliated organs is posterior to the brain. The homology between these and the apical organ of a trochophore larva is discussed. A distinctive longitudinal rod, the chordoid organ, consists of vacuolized cells with circular myofilaments. The organ is comparable to a similar structure in gastrotrichs. In the discussion of the phylogenetic position of Cycliophora among protostomians, important morphological observations that are described in the present study indicate that, despite some dissimilarities, the chordoid larva is a modified trochophore. © 1996 Wiley-Liss, Inc.     

Dwarf male of Symbion pandora (cycliophora)

This study clarifies the identity and development of the male in the life cycle of Symbion pandora. The male is not produced directly by the feeding stage, as previously thought, but arises as a distinct individual from budding cells inside an intermediate stage named the Prometheus larva. The morphology and the development of the two distinct stages are described with light and electron microscopy. Furthermore, the following terminology is suggested to clearly distinguish between the different individuals: 1) the Prometheus larva, which is the free-swimming individual being produced inside the feeding stage; 2) the attached Prometheus larva on the feeding stage, which mostly degenerates following settlement, except for the internal budding cells; and 3) the dwarf male, which is the ciliated, sexually mature stage. The budding cells inside the attached Prometheus larva usually develop two internal dwarf males. Each dwarf male is heavily ciliated and has a well-developed nervous system with a relatively large brain, numerous gland and muscle cells, testis with bundles of sperm, and one penial structure. The male lacks a gut, as in the other free stages in the life cycle of Symbion pandora. This study also indicates that the dwarf male is freed from the attached Prometheus larva. Copulation, which has not been observed yet, probably takes place between a free-swimming male and the female, either while the female is released or afterwards.     

Peculiarities of ultrastructural organization of the metathoracic ganglion in the locustLocusta migratoria larva

In electron microscopic study of structural organization of the thoracic ganglion of the locust larva of the 1st age (1–2 days after hatching), the data on the structure of motoneurons of the 1st nerve, basal and motor neuropil of the larva were obtained. The effector elements of the larval locust CNS are formed rather early and have the structural plan similar to that in adult insects. However, in the larval motoneurons innervating the flight muscles (longitudinal dorsal muscles, wing depressors) the clearly seen features of immaturity of these nervous elements are revealed. Study of the larval ganglion neuropil has shown that the basal neuropil is morphologically formed sufficiently completely as early as in larvae of the first days after hatching. There are shown longitudinal contacts between axons of the ventral neuropil zone, the presence of axons forming theen-passant contacts as well as the synapses with a heterogeneous set of vesicles in the presynaptic area. The presence of the great number of granular vesicles in the basal neuropil of the locust larva may indicate an important role of catecholamines in the early development of the nervous system in the locust larva.     

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.     

Serotonin in the developing stomatogastric system of the lobster,Homarus americanus

We studied the development of the serotonergic modulation of the stomatogastric nervous system of the lobster, Homarus americanus. Although the stomatogastric ganglion (STG) is present early in embryonic development, serotonin immunoreactivity is not visible in the STG until the second larval stage. However, incubation of the STG with exogenous serotonin showed that a serotonin transporter is present in embryonic and early larval stages. Serotonin uptake was blocked by paroxetine and 0% Na⁺ saline. The presence of a serotonin transporter in the embryonic STG suggests that hormonally liberated serotonin could be taken up by the STG, and potentially released as a “borrowed transmitter”. Consistent with a potential hormonal role, serotonin is found in the pericardial organs, a major neurosecretory structure, by midembryonic development. The rhythmic motor patterns produced by embryonic and larval STGs were decreased in frequency by serotonin. Lateral Pyloric (LP) neuron‐evoked excitatory junctional potentials (EJPs) in the embryos and the first larval stage (LI) were larger, slower, and more variable than those in the adult. The amplitude of adult LP neuron‐evoked EJPs was increased more than twofold in serotonin, but in embryos and LI preparations this effect was negligible. In embryos and LI preparations, serotonin increased the occurrence of muscle fiber action potentials and altered the EJP wave‐form. These data demonstrate that serotonin receptors are present in the stomatogastric nervous system early in development, and suggest that the role of serotonin changes from modulation of muscle fiber excitability early in development to enhancement of neurally evoked EJPs in the adult. © 2002 Wiley Periodicals, Inc. J Neurobiol 54: 380–392, 2003     

The Larval Nervous System of Polygordius lacteus Scheinder, 1868 (Polygordiidae,Polychaeta): Immunocytochemical Data

The nervous system of the planktotrophic trochophore larva of Polygordius lacteus has been investigated using antibodies to serotonin (5-HT) and the neuropeptide FMRFamide. The apical ganglion contains three 5-HT-ir neurons, many FMRFamide-ir neurons and a tripartate 5-HT-ir and FMRFamide-ir neuropil. A lateral nerve extends from each side of the apical ganglion across the episphere and the ventral hyposphere, where the two nerves combine to form the paired ventral nerve cord. These nerves have both 5-HT-ir and FMRFamide-ir processes. Three circumferential nerves are associated with the ciliary bands: two prototroch and one metatroch nerve. All contain 5-HT-ir and FMRFamide-ir processes. An oral nerve plexus also contain both 5-HT-ir and FMRFamide-ir processes develops from the metatroch nerve, and an esophageal ring of FMRFamide-ir processes develops in later larval stages. In young stages the ventral ganglion contains two 5-HT-ir and two FMRFamide-ir perikarya; during development the ventral ganglion grows caudally and adds additional 5-HR-ir and FMRFamide-ir perikarya. These are the only perikarya that could be found along the lateral nerve and ventral nerve cord. The telotroch nerve develops from the ventral nerve cord. The 5-HT-ir and FMRFamide-ir part of the nervous system is strictly bilateral symmetric. and much of the system (i.e. apical ganglion, lateral nerves ventral nerve cord, dorsal nerve and oral plexus) is retained in the adult.     

A new species of Lepidodasys (Gastrotricha, Macrodasyida) from Panama with a description of its peptidergic nervous system using CLSM, anti-FMRFamide and anti-SCPB

A new species of Lepidodasys (Gastrotricha: Macrodasyida: Lepidodasyidae) is described from sublittoral sediments in the Bocas del Toro archipelago in Panama and represents the first species of Lepidodasyidae described from the Caribbean. The new species possesses keeled scales that form a crossed-helical pattern across its dorsal and lateral surfaces and ventral scales that form a herringbone pattern between and lateral to the ciliary columns. A bilateral pair of three ventral adhesive tubes at the posterior end further differentiates this new species from its seven congeners. A confocal laser scanning microscope examination of the nervous system using antibodies to small cardioactive peptide B (SCP_B) and FMRFamides reveals a dumbbell-shape cerebral ganglion, paired pharyngeal neurites and paired posterior nerve cords. Expression patterns of immunoreactivity to both classes of neuropeptides show a high degree of similarity. Only within the lateral somata of the cerebral ganglion and a single median pharyngeal neurite is there a difference in immunoreactivity to FMRFamide (positive) compared to SCP_B (negative). Results from this investigation reveal that neuropeptides, among other neuronal markers, might provide phylogenetically informative characters in macrodasyidan gastrotrichs, especially regarding the topology of the cerebral ganglion.     

Serotonin immunoreactivity in the nervous system of the free‐swimming larvae and sessile adult females of Stephanoceros fimbriatus (Rotifera: Gnesiotrocha)

Unlike most rotifers (Rotifera), which are planktonic and direct developers, many gnesiotrochan rotifers (Monogononta: Gnesiotrocha) are sessile and have indirect development. Few details exist on larval metamorphosis in most gnesiotrochans, and considering the drastic transformation that takes place at metamorphosis—the replacement of the ciliated corona with a new head that bears ciliated tentacles (the infundibulum)—it is perhaps surprising that there are limited data on the process. Here, we document part of this metamorphosis by examining the presence and distribution of neurons with serotonin immunoreactivity in the nervous system of both planktonic larvae and sessile adult females. Using antibodies against serotonin combined with confocal laser‐scanning microscopy (CLSM) and 3D reconstruction software, we mapped the immunoreactive cell bodies and neurites in both life stages and found that relatively few changes occurred during metamorphosis. The larvae possessed a total of eight perikarya with serotonergic immunoreactivity (5HT‐IR) in the brain, with at least two pairs of perikarya outside the brain in the region of the corona. Cells with 5HT‐IR in the brain innervated the larval corona and also sent neurites to the trunk via the nerve cords. During metamorphosis, the corona was replaced by the infundibulum, which emerged from the larval mouth to become the new functional head. This change led to a posterior displacement of the brain and also involved the loss of 5HT‐IR in the lateral brain perikarya and the gain of two perikarya with 5HT‐IR in the anterior brain region. The innervation of the anterior end was retained in the adult; neurites that extended anteriorly to the mouth of the larva formed a distinct neural ring that encircled the infundibulum after metamorphosis. Significantly, there was no innervation of the infundibular tentacles by neurites with 5HT‐IR, which suggests that ciliary control is unlikely to be modulated by serotonin within the tentacles themselves.     

Detection of negative buoyancy in the phantom larva, Chaoborus americanus

Fourth instar larvae of Chaoborus americanus regain neutral buoyancy after receiving weights or under added hydrostatic pressure by increasing the size of gas-filled tracheal sacs. The rate and degree of the response are directly related to the velocity of vertically flowing current experienced by the larva. Weighted larvae showing the more rapid sinking rates also show the more rapid adjustment rates. The usual response disappears when weighted larvae are suspended but reappears when water is caused to flow upwards past the stationary larva. The degree of sac expansion under any increment in hydrostatic pressure may be reduced by decreasing larval sinking rates with polyethylene floats. These results support the hypothesis of previous workers that water currents initiate the process of buoyancy adjustment.     

FMRfamide-like immunoreactivity in the ventral nerve cord of the larval eastern spruce budworm,Choristoneura fumiferana (clemens) (Lepidoptera : Tortricidae)

Distribution of FMRFamide-like immunoreactivity was examined in the larval ventral nerve cord of the eastern spruce budworm, Choristoneura fumiferana (Lepidoptera : Tortricidae). Indirect immunofluorescent methods revealed the existence of 3 groups of FLI neurons in each ganglion. The neurons are distributed in a bilaterally symmetrical fashion at the anterodorsal, midlateral and posteroventral regions of the ganglia. There are 4 FMRFamide-like immunoreactive fiber tracts on the dorsal surface of the ganglia to which the anterodorsal FLI neurons project ipsilaterally, while the midlateral pair projects both ipsi-, and contralaterally. The last abdominal ganglion (AG8) has 4 additional pairs of FLI neurons; and axons from some of these extend into the median abdominal nerve, which suggests some role for this neuropeptide in the control of posterior structures of the larva.     

Development of the nervous system in Phoronopsis harmeri (Lophotrochozoa, Phoronida) reveals both deuterostome- and trochozoan-like features

Background

Inferences concerning the evolution of invertebrate nervous systems are often hampered by the lack of a solid data base for little known but phylogenetically crucial taxa. In order to contribute to the discussion concerning the ancestral neural pattern of the Lophotrochozoa (a major clade that includes a number of phyla that exhibit a ciliated larva in their life cycle), we investigated neurogenesis in Phoronopsis harmeri, a member of the poorly studied Phoronida, by using antibody staining against serotonin and FMRFamide in combination with confocal microscopy and 3D reconstruction software.

Results

The larva of Phoronopsis harmeri exhibits a highly complex nervous system, including an apical organ that consists of four different neural cell types, such as numerous serotonin-like immunoreactive flask-shaped cells. In addition, serotonin- and FMRFamide-like immunoreactive bi- or multipolar perikarya that give rise to a tentacular neurite bundle which innervates the postoral ciliated band are found. The preoral ciliated band is innervated by marginal serotonin-like as well as FMRFamide-like immunoreactive neurite bundles. The telotroch is innervated by two neurite bundles. The oral field is the most densely innervated area and contains ventral and ventro-lateral neurite bundles as well as several groups of perikarya. The digestive system is innervated by both serotonin- and FMRFamide-like immunoreactive neurites and perikarya. Importantly, older larvae of P. harmeri show a paired ventral neurite bundle with serial commissures and perikarya.

Conclusions

Serotonin-like flask-shaped cells such as the ones described herein for Phoronopsis harmeri are found in the majority of lophotrochozoan larvae and therefore most likely belong to the ground pattern of the last common lophotrochozoan ancestor. The finding of a transitory paired ventral neurite bundle with serially repeated commissures that disappears during metamorphosis suggests that such a structure was part of the ??ur-phoronid?? nervous system, but was lost in the adult stage, probably due to its acquired sessile benthic lifestyle.     

New light on the enigmatic Xenoturbella (phylum uncertain): ontogeny and phylogeny

Xenoturbella is an enigmatic animal that is merely a ciliated bag with epithelial epidermis and gastrodermis, a subepidermal nerve plexus and a ventral mouth, but without an anus or any distinct organs. It is marine, free living, and up to 4 cm long. Its simplicity in organization has led to diverse interpretations during the last 50 years: as an acoelomorph flatworm, a paedomorphic holothurian or enteropneust, or a unique representative of a plesiomorphic phylum. I report here the previously unknown embryology of Xenoturbella that unequivocally corroborates a bivalve relationship and thus once and for all dismisses the potential new phylum. The simplicity of the adult Xenoturbella is due to neither plesiomorphy nor paedomorphy. It is caused by metamorphosis from a trochophore larva of molluscan type with a defined organ system, including a concentrated nervous system with ganglia, to an adult without any defined organs.     

Localization of a SALMFamide Neuropeptide in the Larval Nervous System of the Sand Dollar Dendraster excentricus

Using immunocytochemical methods we describe the localization of serotonin and the SALMFamide peptide, S1 (GFNSALMFamide), during embryonic and larval development of the echinoid Dendraster excentricus. Anti-SI immunoreactivity first appears in the apical ganglion in late gastrulae at the same time as anti-serotonin immunoreactivity. Initially, anti-S1 immunoreactivity is restricted to fibres of the neuropile, but in later feeding stages, cell bodies are also immunoreactive. Anti-S1 immunoreactivity appears as 2–4 cells in the oral ganglion of early prism stage larvae, whereas anti-serotonin immunoreactivity does not occur in the oral ganglion until the 8-arm stage. Anti-S1 immunoreactivity also occurs in diffuse fibres in the oesophagus and in a single fibre encircling the pyloric sphincter of the gut. A reticular network associated with the apical surface of the epithelial cells of the vestibule of the adult rudiment was anti-S1 immunoreactive. In double-labelling experiments, anti-serotonin and anti-S1 immunoreactivity co-localize in the neuropile of the apical ganglion. The distribution of S1, in association with putative sensory cells in the apical and oral ganglia and with muscles of the oesophagus and gut, suggests S1 may have diverse functions in the larval nervous system. The distribution of anti-S1 immunoreactivity in echinoid embryos and larvae supports the proposal that SALMFamide-like peptides are widely shared in echinoderms and potentially have a fundamental role in neural function.