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

Introduction

Acoels 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.

Results

The 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.

Conclusions

The 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.     

Interhemispheric connections through the pallial commissures in the brain of Podarcis hispanica and Gallotia stehlinii (Reptilia,Lacertidae)

The cells-of-origin and the mode and site of termination of the interhemispheric connections passing through the anterior and posterior pallial commissures in the telencephalon of two lizards (Podarcis hispanica and Gallotia stehlinii) were investigated by studying the anterograde and retrograde transport of unilaterally injected horseradish peroxidase. The commissural projections arise mainly from pyramidal cells in the medial, dorsomedial, and dorsal cortices (medial subfield). Additionally some non-pyramidal neurons in the medial and dorsal cortices contribute to the commissural system. Medial cortex neurons project to the contralateral anterior septum through the anterior pallial commissure. The dorsomedial cortex projects contralaterally via the anterior pallial commissure to the dorsolateral septum and to the medial, dorsomedial, and dorsal cortices. The projection to the medial cortex terminates in two bands at the inner and outer border, respectively, of the cell layer; the projection to the dorsomedial and dorsal cortex ends in a zone in layer 1 which previously has been described to be Timm-negative, and in a diffuse band in the inner half of layer 3. The medial subfield of the dorsal cortex projects through the anterior pallial commissure to the dorsomedial and dorsal cortices with a similar pattern of termination to that found for the dorsomedial cortex. The posterior pallial commissure contains only the projections from the ventral cortex to its contralateral counterpart and to the ventral part of the caudal medial cortex. The similarities found between this commissural system and the mammalian hippocampal interhemispheric connections are discussed.     

The nervous system of <Emphasis Type="Italic">Convolutriloba</Emphasis> (Acoela) and its patterning during regeneration after asexual reproduction

We describe the serotonergic and cholinergic nervous system of the asexually reproducing acoel Convolutriloba longifissura Bartolomaeus & Balzer, 1997 by means of immunohistochemistry, conventional histochemistry and transmission electron microscopy. Immunocytochemical staining for serotonin revealed neurons in the brain, in a pair of ventral main longitudinal cords, in two pairs of smaller dorsal longitudinal nerve cords, and in a submuscular nerve net. The brain comprises a ventral-anterior commissure and a less intensely stained dorsal commissure joined together by connectives into a three-ringed scaffold from which the longitudinal nerves extend. We followed the regeneration of the serotonergic part of the nervous system up to the second day after fission. Within this time period, the offspring reestablished bilateral symmetry in the nervous system and developed full motor control. The presence of aminergic cell bodies associated with the main lateral nerve cords of C. longifissura shows that the acoelan nervous system is more similar to that of other platyhelminths (triclads, rhabditophorans) than previously assumed. The presence of serotonergic cell bodies along the main nerve cord correlates with the capacity for asexual reproduction via fissioning. We also describe the single fission mode of C. hastifera Winsor 1990, which brings the modes of asexual reproduction employed by members of the Convolutrilobinae to three.     

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.     

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.     

The Hox gene complement of acoel flatworms, a basal bilaterian clade

Several molecular data sets suggest that acoelomorph flatworms are not members of the phylum Platyhelminthes but form a separate branch of the Metazoa that diverged from all other bilaterian animals before the separation of protostomes and deuterostomes. Here we examine the Hox gene complement of the acoel flatworms. In two distantly related acoel taxa, we identify only three distinct classes of Hox gene: an anterior gene, a posterior gene, and a central class gene most similar to genes of Hox classes 4 and 5 in other Bilateria. Phylogenetic analysis of these genes, together with the acoel caudal homologue, supports the basal position of the acoels. The similar gene sets found in two distantly related acoels suggest that this reduced gene complement may be ancestral in the acoels and that the acoels may have diverged from other bilaterians before elaboration of the 8- to 10-gene Hox cluster that characterizes most bilaterians.     

Architecture of the nervous system in two Dactylopodola species (Gastrotricha, Macrodasyida)

Immunohistochemical stainings have become standard tools to describe the nervous system, but usually only singular or few markers are used and consequently show only subsets of neurons within the nervous system. We investigated two species of Dactylopodola (Gastrotricha, Macrodasyida) with a broad set and combination of markers, to represent the nervous system in a more holistic approach. We suggest that markers for both neurotubuli (tubulin) and neurotransmitters (e.g. serotonin, FMRF-amides, histamine) should be used. Combinations with markers for the musculature (phalloidin) and nuclei (propidiumiodide or other markers) help to reveal spatial patterns and when used with TEM can provide a more precise picture of the spatial relationships of particular nerves. Species of Dactylopodola have a brain consisting of a solid dorsal commissure and a fine ventral commissure. Cell somata of brain cells are arranged lateral to the dorsal commissure and form a dumbbell-like brain. Additionally, projections into the head region, head sensory organs, one pair of lateroventral nerve cords with three commissures and stomatogastric nerves are described. Obviously, some longitudinal transmitter-specific fibres run in parallel to the main longitudinal nerve and represent additional longitudinal fibres. In comparison with the nervous system architecture of other gastrotrich species and that of different bilaterian animals it is speculated that the gastrotrich nervous system retains several ancestral features, such as being commissural and not a compact brain.     

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

Bery, A. and Martínez, P. 2010. Acetylcholinesterase activity in the developing and regenerating nervous system of the acoel Symsagittifera roscoffensis. —Acta Zoologica (Stockholm) 92 : 383–392. The use of the cholinergic system is widespread in the animal kingdom. It controls different processes, including reproduction and neural transmission. However, its evolutionary history is not yet well understood. For instance, the role played 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 and regeneration of the acoel flatworm Symsagittifera roscoffensis, using acetylcholinesterase (AchE) activity as a marker. In this species, AchE activity is observed at all developmental stages, including in the early embryos. The juvenile and adult patterns reveal the presence of a complex nervous system that includes three pairs of longitudinal neurite bundles, which are connected to an anterior centralized mass of neurons and neural processes formed by two pairs of connectives and four commissures. The power of the technique also allows the detection of newly born neurons as they are incorporated into the growing nervous system (during regeneration).     

Correlation of fluorescence and electron microscopy of F-actin-containing sensory cells in the epidermis of Convoluta pulchra (Platyhelminthes: Acoela)

Phalloidin‐stained whole mounts of acoel turbellarians show brightly fluorescing club‐shaped structures distributed over the epidermis and concentrated especially at the anterior and posterior tips of the body. By correlating electron micrographic images and fluorescence images of Convoluta pulchra, these structures can be seen to be sensory receptors with a central cilium surrounded by a collar of microvilli. The other candidate for showing fluorescence in the epidermis, namely gland necks, can be ruled out since their distribution is too dense to resemble the distribution of the fluorescent structures seen here. The collared sensory receptors were inserted between epidermal cells, and each bore a central cilium surrounded by a collar of 6–18 microvilli and an additional centrally positioned 2–7 microvilli of which 2 or 3 were associated with a modified rootlet called the swallow’s nest. Confocal scanning laser microscopy resolved the core of actin filaments within the microvilli of the collar and their rootlet‐like connections to the base of the sensory cell. Such receptors could also be identified by fluorescence microscopy in several other species of acoel turbellarians.     

The brain structure of selected trypanorhynch tapeworms

The brain architecture in four species of tapeworms from the order Trypanorhyncha has been studied. In all species, the brain consists of paired anterior and lateral lobes, and an unpaired central lobe. The anterior lobes connect by dorsal and ventral semicircular commissures; the central and lateral lobes connect by a median and an X-shaped crisscross commissure. In the center of the brain, five well-developed compact neuropils are present. The brain occupies a medial position in the scolex pars bothrialis. The ventral excretory vessels are situated outside the lateral lobes of the brain; the dorsal excretory vessels are located inside the brain and dorsal to the median commissure. The brain gives rize four anterior proboscis nerves and four posterior bulbar nerves with myelinated giant axons (GAs). The cell bodies of the GAs are located within the X-commissure and in the bulbar nerves. Highly developed serotonergic neuropils are present in the anterior and lateral lobes; numerous 5-HT neurons are found in the brain lobes including the central unpaired lobe. The X-cross commissure consists of the α-tub-immunoreactive and 5-HT-IR neurites. Eight ultrastructural types of neurons were found in the brain of the three species investigated. In addition, different types of synapses were present in the neuropils. Glial cells ensheath the brain lobes, the neuropils, the GAs, and the bulbar nerves. Glia cell processes form complex branching patterns of thin cytoplasmic sheets sandwiched between adjacent neural processes and filling the space between neurons. Multilayer myelin-like envelopes and a mesaxon-like structure have been found in Trypanorhyncha nervous system. We compared the brain architecture of Trypanorhyncha with that of an early basal cestode taxon, that is, Diphyllobothriidea, and present a hypothesis about the homology of the anterior brain lobes in order Trypanorhyncha; and the lateral lobes and median commissure are homologous brain structures within Eucestoda.     

Walking on inclines: how do desert ants monitor slope and step length

Background

In order to increase the weak database concerning the organogenesis of Acoela – a clade regarded by many as the earliest extant offshoot of Bilateria and thus of particular interest for studies concerning the evolution of animal bodyplans – we analyzed the development of the musculature of Symsagittifera roscoffensis using F-actin labelling, confocal laserscanning microscopy, and 3D reconstruction software.

Results

At 40% of development between egg deposition and hatching short subepidermal fibres form. Muscle fibre development in the anterior body half precedes myogenesis in the posterior half. At 42% of development a grid of outer circular and inner longitudinal muscles is present in the bodywall. New circular muscles either branch off from present fibres or form adjacent to existing ones. The number of circular muscles is higher than that of the longitudinal muscles throughout all life cycle stages. Diagonal, circular and longitudinal muscles are initially rare but their number increases with time. The ventral side bears U-shaped muscles around the mouth, which in addition is surrounded by a sphincter muscle. With the exception of the region of the statocyst, dorsoventral muscles are present along the entire body of juveniles and adults, while adults additionally exhibit radially oriented internal muscles in the anterior tip. Outer diagonal muscles are present at the dorsal anterior tip of the adult. In adult animals, the male gonopore with its associated sexual organs expresses distinct muscles. No specific statocyst muscles were found. The muscle mantles of the needle-shaped sagittocysts are situated along the lateral edges of the animal and in the posterior end close to the male gonopore. In both juveniles and adults, non-muscular filaments, which stain positively for F-actin, are associated with certain sensory cells outside the bodywall musculature.

Conclusion

Compared to the myoanatomy of other acoel taxa, Symsagittifera roscoffensis shows a very complex musculature. Although data on presumably basal acoel clades are still scarce, the information currently available suggests an elaborated musculature with longitudinal, circular and U-shaped muscles as being part of the ancestral acoel bodyplan, thus increasing the possibility that Urbilateria likewise had a relatively complicated muscular ground pattern.     

The neuroactive substances and associated muscle system in Rhipidocotyle campanula (Digenea,Bucephalidae) from the intestine of the pike Esox lucius

We report about the muscular system and the serotonergic and FMRFamidergic components of the nervous system of the Bucephalidae trematode, Rhipidocotyle campanula, an intestinal parasite of the pike. We use immunocytochemical methods and confocal scanning laser microscopy (CLSM). The musculature is identified by histochemical staining with fluorescently labeled phalloidin. The body wall musculature of R. campanula contains three layers of muscle fibres – the outer thin circular, intermediate longitudinal and inner diagonal muscle fibres running in two opposite directions. The digestive system of R. campanula possess of a well-developed musculature: radial, longitudinal and circular muscle elements are detected in the pharynx, circular and longitudinal muscle filaments seen in the oesophagus, and longitudinal and the circular muscle fibres were found in the intestinal wall. Specific staining indicating the presence of actin muscle filaments occurs in the cirrus sac localized in the posterior body region. The frontal region of anterior attachment organ, the rhynchus, in R. campanula is represented by radial muscle fibres. The posterior part of the rhynchus comprise of radial muscles forming the organ's wall, and several strong longitudinal muscle bundles. Serotonergic and FMRFamidergic structures are detected in the central and peripheral compartments of the nervous system of R. campanula, that is, in the paired brain ganglia, the brain commissure, the longitudinal nerve cords, and connective nerve commissures. The innervations of the rhynchus, pharynx, oesophagus and distal regions of the reproductive system by the serotonergic and FMRFamidergic nervous elements are revealed. We compare our findings obtained on R. campanula with related data for other trematodes.     

The midline glial cells are required for regionalization of commissural axons in the embryonic CNS of Drosophila

 The ventral nerve cord of arthropods is characterised by the organisation of major axon tracts in a ladder-like pattern. The individual neuromeres are connected by longitudinal connectives whereas the contra-lateral connections are brought about through segmental commissures. In each neuromere of the embryonic central nervous system (CNS) of Drosophila an anterior and a posterior commissure is found. The development of these commissures requires a set of neurone-glia interactions at the midline. Here we show that both the anterior as well as the posterior commissures are subdivided into three axon-containing regions. Electron microscopy of the ventral nerve cord of mutations affecting CNS midline cells indicates that the midline glial cells are required for this subdivision. In addition the midline glial cells appear required for a crossing of commissural growth cones perpendicular to the longitudinal tracts, since in mutants with defective midline glial cells commissural axons frequently cross the midline at aberrant angles. Received: 6 July 1997 / Accepted: 27 August 1997     

The central and peripheral nervous system of <Emphasis Type="Italic">Cephalodiscus gracilis</Emphasis> (Pterobranchia,Deuterostomia)

Nervous systems are important in assessing interphyletic phylogenies because they are conservative and complex. Regarding nervous system evolution within deuterostomes, two contrasting hypotheses are currently discussed. One that argues in favor of a concentrated, structured, central nervous system in the last common ancestor of deuterostomes (LCAD); the other reconstructing a decentralized nerve net as the nervous system of the LCAD. Here, we present a morphological analysis of the nervous system of the pterobranch deuterostome Cephalodiscus gracilis Harmer, 1905 based on transmission electron microscopy, confocal laser scanning microscopy, immunohistochemistry, and computer-assisted 3D reconstructions based on complete serial histological sections. The entire nervous system constitutes a basiepidermal plexus. The prominent dorsal brain at the base of the mesosomal tentacles contains an anterior concentration of serotonergic neurons and a posterior net of neurites. Predominant neurite directions differ between brain regions and synapses are present, indicating that the brain constitutes a centralized portion of the nervous system. Main structures of the peripheral nervous system are the paired branchial nerves, tentacle nerves, and the ventral stalk nerve. Serotonergic neurites are scattered throughout the epidermis and are present as concentrations along the anterior border of the branchial nerves. Serotonergic neurons line each tentacle and project into the brain. We argue that the presence of a centralized brain in C. gracilis supports the hypothesis that a nerve center was present in the LCAD. Moreover, based on positional and structural similarity, we suggest that the branchial nerves in C. gracilis could be homologous to branchial nerves in craniates, a hypothesis that should be further investigated.     

Monograph of the solenofilomorphidae (Turbellaria: Acoela)

Morphological studies of eleven new species in the family permitted reconstruction of progressive evolutionary lines, and this strongly implies progressive evolutionary lines for other acoela. Solenofilomorpha funilis n. sp., Myopea crassula n. g. n. sp., M. latafaucium n. sp., Fusantrum rhammiphorum n. g. n. sp. and Endocincta punctata n. g. n. sp. are described from the coast of South Carolina, U.S.A., and S. guaymensis n. sp. is described from the Gulf of California. Five additional unnamed species from the U. S. Pacific Coast, Sweden and Tunisia are briefly described. Two species groupings were based on correlation between two different arrangements of pharynx muscles and trio arrangements of antrum muscles and further correlations with lesser characters. The membership of the two groupings in a single family make it clear that evolution proceeded by addition of structures to a less complex common ancestor. Functional and behavioral considerations support the evolutionary path drawn from morphology. Distinctive statocyst structure, ciliary interconnections and sperm morphology shared by acoels show them to have a common ancestry, but clearly different pharynges and male structures cannot be consistent with evolution by reductions. A common ancestor of low complexity has probably given rise to several partly parallel evolutionary lines which together form the structurally diverse Acoela. The history of acoel systematics is also briefly summarized.     

The sensory innervation of the pineal organ in the lizard,Lacerta viridis,with remarks on its position in the trend of pineal phylogenetic structural and functional evolution

Summary The sensory innervation of the pineal organ of adult Lacerta viridis has been investigated. Some specimens of Lacerta muralis lillfordi were also used. In the pineal epithelium, a small number of nerve cell pericarya of a sensory type are present. They lie either solitary or in small clusters close to the basement membrane. The axons originating from the nerve cell bodies, i. e. the pineal sensory nerve fibers, first course in the intraepithelial nerve fiber layer which is only locally present and contains a restricted number of unmyelinated fibers. In Lacerta viridis, the pineal fibers generally leave the epithelium at the proximal part of the organ proper. They then form small bundles which run along the outer surface of the basement membrane in the leptomeningeal connective tissue covering. At the proximal end of the pineal stalk the single bundles assemble constituting the pineal nerve. In Lacerta muralis the fibers leave the pineal epithelium at the proximal end of the stalk running farther down within the epithelium. Many fibers become myelinated after leaving the pineal epithelium. The pineal nerve runs ventralward in the midplane just caudal to the habenular commissure to which no fibers are given off. Continuing their ventralward course between the habenular commissure and the rostral end of the posterior commissure which is traversed by some of them, the pineal fibers reach the dorsal border of the subcommissural organ. Small separate aberrant pineal bundles traverse the posterior commissure at various more caudal levels. Having reached the dorsal border of the subcommissural organ, part of the pineal fibers continue their ventralward course directly running along the lateral sides of this organ to reach the periventricular nerve fiber layer lateral and ventral to it. A restricted number of fibers first turns in a caudal direction running between the base of the posterior commissure and the base of the subcommissural organ before turning ventralward to reach the periventricular layer. Most probably, pineal fibers do neither join the posterior commissural system nor innervate the subcommissural organ. Once having reached the periventricular layer, some pineal fibers curve in a rostral direction while others, before doing so, send a collateral in a caudal direction. Both, the main fibers and the collaterals, contribute to the formation of the periventricular layer. The sites of termination of the pineal fibers could not be ascertained.From the presence of intraepithelial sensory nerve cell bodies and from literature data on the ultrastructure of pineal neurosensory cells it is concluded that the adult pineal organ of Lacerta has a, although rudimentary, (photo)sensory function. The demonstration by our guest-worker Dr. W. B. Quay, of the intraepithelial presence of a tryptamine compound, probably serotonin, points, moreover, to a secretory function of this organ.In adult Lacerta a well-developed parietal nerve connects the parietal eye with the left lateral habenular nucleus. It traverses the habenular commissure.In gratitude and with admiration this paper is dedicated to Prof. Berta Scharrer and to the memory of Prof. Ernst Scharrer.     

Neuromuscular development in Novocrania anomala: evidence for the presence of serotonin and a spiralian-like apical organ in lecithotrophic brachiopod larvae

SUMMARY The phylogenetic position of Brachiopoda remains unsettled, and only few recent data on brachiopod organogenesis are currently available. In order to contribute data to questions concerning brachiopod ontogeny and evolution we investigated nervous and muscle system development in the craniiform (inarticulate) brachiopod Novocrania anomala . Larvae of this species are lecithotrophic and have a bilobed body with three pairs of dorsal setal bundles that emerge from the posterior lobe. Fully developed larvae exhibit a network of setae pouch muscles as well as medioventral longitudinal and transversal muscles. After settlement, the anterior and posterior adductor muscles and delicate mantle retractor muscles begin to form. Comparison of the larval muscular system of Novocrania anomala with that of rhynchonelliform (articulate) brachiopod larvae shows that the former has a much simpler muscular organization. The first signal of serotonin-like immunoreactivity appears in fully developed Novocrania anomala larvae, which have an apical organ that consists of four flask-shaped cells and two ventral neurites. These ventral neurites do not stain positively for the axonal marker α-tubulin in the larval stages. In the juveniles, the nervous system stained by α-tubulin is characterized by two ventral neurite bundles with three commissures. Our data are the first direct proof for the presence of an immunoreactive neurotransmitter in lecithotrophic brachiopod larvae and demonstrate the existence of flask-shaped serotonergic cells in the brachiopod larval apical organ, thus significantly increasing the probability that this cell type was part of the bauplan of the larvae of the last common lophotrochozoan ancestor.