Distribution and Ultrastructure of Ciliated Sensory Receptors in the Posterior Mantle Epithelium of Dentalium rectius (Mollusca,Scaphopoda)

The scaphopod mantle cavity opens posteriorly via the pavilion, a siphon-like extension of the posterior mantle through which the respiratory currents pass. The pavilion was examined for ciliated sensory cells in Dentalium rectius Carpenter, 1865, using scanning and transmission electron micropscopy. Three types of sensory receptor were distinguished on the basis of number, length and ultrastructure of the associated cilia. Receptors with 2–5 cilia of ? 1.7 μm length lined the pavilion edge. A second type, possessing 1–2 cilia, ? 8.2 μm in length, was found throughout the internal and on part of the external surface of the pavilion. The third receptor type consisted of a rigid bundle of 16–40 cilia with a length of ? 14.4 μm, and was present close to the periphery and at the base of the pavilion near the entrance to the mantle cavity. The structure and distribution of these cells are similar to peripheral chemo- and mechanoreceptors which sample respiratory currents and the surrounding environment in other molluscs, but they may assume a greater functional significance in scaphopods due to the absence of an osphradium in this class.     

The protonephridial system of the tusk shell, Antalis entalis (Mollusca, Scaphopoda)

The development and microanatomy of the protonephridial system in larvae and postmetamorphic juveniles of Antalis entalis (Dentaliidae) have been examined by means of a semithin serial sectioning and reconstruction technique. One late larval stage has been additionally examined by transmission electron microscopy. The protonephridium appears during larval development and is reduced in the juvenile approximately 13 days after metamorphosis. This is the first unambiguous evidence of a protonephridium in a postlarval mollusc. When fully developed the protonephridium is unique in consisting of two cells only, a terminal cell (=cyrtocyte) and a duct-releasing cell with glandular appearance. The polyciliary terminal cell has several distinct ultrafiltration sites, resembling conditions in bivalve protonephridia. The large duct-releasing cell shows a very large nucleus probably reflecting polyploidy. Its basal infoldings and many mitochondria suggest metabolic activity, the cytoplasm is characterised by many distinct granules. The unique features of the scaphopod protonephridial system are compared with available data on the protonephridia of other molluscan classes. The finding gives additional evidence that protonephridia belong to the ground pattern of the Mollusca. Accepted: 22 January 2001     

Phylogenetic relationships among families of the Scaphopoda (Mollusca)

Phylogenetic relationships among families in the molluscan class Scaphopoda were analysed using morphological characters and cladistic parsimony methods. A maximum parsimony analysis of 34 discrete characters, treated as unordered and equally weighted, from nine ingroup terminal taxa produced a single most parsimonious tree; supplementary analyses of tree length frequency distribution and Bremer support indices indicate a strong phylogenetic signal from the data and moderate to minimally supported clades. The traditional major division of the class, the orders Dentaliida and Gadilida, is supported as both taxa are confirmed as monophyletic clades. Within the Dentaliida, two clades are recognized, the first comprised of the families Dentaliidae and Fustiariidae, the second of the Rhabdidae and Calliodentaliidae; together, these groups comprise a third clade, which has the Gadilinidae as sister. Within the Gadilida, a nested series of relationships is found among [Entalinidae, [Pulsellidae, [Wemersoniellidae, Gadilidae]]]. These results lend cladistic support to earlier hypotheses of shared common ancestry for some families, but are at variance with other previous hypotheses of evolution in the Scaphopoda. Furthermore, analysis of constituent Gadilinidae representatives provide evidence for paraphyly of this family. The relationships supported here provide a working hypothesis that the development of new characters and greater breadth of taxonomic sampling can test, with a suggested primary goal of establishing monophyly at the family level.     

Muscle development in Antalis entalis (Mollusca,Scaphopoda) and its significance for scaphopod relationships

We applied fluorescence staining of F-actin, confocal laser scanning microscopy, as well as bright-field light microscopy, SEM, and TEM to examine myogenesis in larval and early juvenile stages of the tusk-shell, Antalis entalis. Myogenesis follows a strict bilaterally symmetrical pattern without special larval muscle systems. The paired cephalic and foot retractors appear synchronously in the early trochophore-like larva. In late larvae, both retractors form additional fibers that project into the anterior region, thus enabling retraction of the larval prototroch. These fibers, together with the prototroch, disappear during metamorphosis. The anlagen of the putative foot musculature, mantle retractors, and buccal musculature are formed in late larval stages. The cephalic captacula and their musculature are of postmetamorphic origin. Development of the foot musculature is dramatically pronounced after metamorphosis and results in a dense muscular grid consisting of outer ring, intermediate diagonal, and inner longitudinal fibers. This is in accordance with the proposed function of the foot as a burrowing organ based on muscle-antagonistic activity. The existence of a distinct pair of cephalic retractors, which is also found in basal gastropods and cephalopods, as well as new data on scaphopod shell morphogenesis and recent cladistic analyses, indicate that the Scaphopoda may be more closely related to the Gastropoda and Cephalopoda than to the Bivalvia.     

Comparative neuroanatomy of Caudofoveata, Solenogastres, Polyplacophora, and Scaphopoda (Mollusca) and its phylogenetic implications

The nervous system of invertebrates is considered to be a very conservative organ system and thus can be helpful to elucidate questions of phylogenetic relationships. Up to now, comparative neuroanatomical studies have been mainly focused on arthropods, where in-depth studies on major brain structures are abundant. In contrast, except for Gastropoda and Cephalopoda, the nervous system of representatives of the second largest phylum of invertebrates, the Mollusca, is as yet hardly investigated. We therefore initiated an immunohistochemical survey to contribute new neuroanatomical data for several molluscan taxa, especially the lesser known Caudofoveata, Solenogastres, Polyplacophora, and Scaphopoda, focusing on the cellular architecture and distribution of neurotransmitters in the brain. Antisera against the widespread neuroactive substances FMRFamide and serotonin were used to label subsets of neurons. Both antisera were additionally used in combination with acetylated ??-tubulin and the nuclear marker DAPI. This enables us to describe the morphology of the nervous system at a fine resolution and to compare its cellular architecture between different species of one taxon, as well as between different taxa of mollusks. On the basis of these results, the nervous system of caudofoveates seems to be most highly derived within the so-called basal (non-conchiferan) mollusks, and a monophyly of a clade Aplacophora could not be confirmed. In general, the brain as well as the remaining nervous system of the molluscan taxa investigated shows a great variability, suggesting a deep time origin of the diversification of this prominent protostome clade.     

The phylogeny and classification of Scaphopoda (Mollusca): an assessment of current resolution and cladistic reanalysis

The first cladistic analysis of phylogeny in the class Scaphopoda (Steiner 1992a,1996) examined relationships among family and selected sub-family taxa using morphological data. A preferred/ consensus tree of relationships illustrated monophyly of the orders Dentaliida and Gadilida, partial resolution among dentaliid families, and complete resolution among gadilid taxa. However, several alternative replications of the analysis, including use of a revised data matrix, did not produce the reported tree number or level of resolution; in all cases, monophyly of the Dentaliida was not supported by strict consensus of resultant parsimonious trees. Reanalysis, using unordered characters and outgroup rooting, only clearly resolves monophyly of the Gadilida and the sister relationship of the Entalinidae with the remaining gadilid families. These analyses emphasize the need for more comparative data and thorough parsimony analysis in scaphopod cladistic phylogenetics, as relationships in this class are still some way from resolution.     

Fine structure of the kidney and characterization of secretory products inDentalium rectius (Mollusca,Scaphopoda)

Summary The ultrastructure of the scaphopod kidney and secretory product composition is described, for the first time, inDentalium rectius. The kidney epithelium consists of two primarily secretory cell types. The first exhibits extensive vacuolation, and scattered granules are formed within the vacuolar space by a process of surface accretion; the incorporation of glycogen particles in this process is associated with very fine, electronopaque threads which radiate from the granule. The second cell type possesses granules enclosed individually within secretory vesicles, and intermediate stages in their growth are characterized by needle-like crystals on the granule surface. The secretory vesicles in some cases coalesce to form a large central vacuole filled with granules. This cell type possesses an apical membrane with sparse microvilli, which may indicate a secondary reabsorptive capacity. Granules in both cell types show a concentric ring ultrastructure, and are composed primarily of calcium phosphate with a small amount of zinc; there is also an organic component of protein, mucopolysaccharide and a large amount of glycogen. Ultrastructural and histochemical observations indicate a lysosomal origin for the granules, although granules of the second cell type develop intracellularly to a greater extent than those of the first. All granules are extruded into the kidney lumen by a process of merocrine secretion prior to release into the mantle cavity via an externally ciliated, muscular excretory pore.     

The chiton gill: Ultrastructure in Chiton olivaceus (Mollusca,Polyplacophora)

Gills of Chiton olivaceus, a primitive mollusc, are relatively simple in their structure and ultrastructure but are well adapted to a life in the intertidal zone. In contrast to some other molluscs, there is no differentiation of the gill epithelium into functional regions other than respiratory ones. Ciliation of the epithelium in certain areas may optimize water flow from the outer to the inner part of the mantle cavity. The hemolymph sinuses are oriented so that hemolymph flows in the opposite direction. Interstitial cells link epithelial cells with nerve endings. Muscle cells as well as the collagenous matrix in the connective tissue differ within the main gill axis and the lateral lamellae. The life cycle of immunoactive cells within the connective tissue and the hemolymph is described.     

The expression of an engrailed protein during embryonic shell formation of the tusk-shell, Antalis entalis (Mollusca, Scaphopoda)

SUMMARY This study presents the first detailed account of the larval and early post-metamorphic development of a scaphopod species, Antalis entalis , since 1883. Special reference is given to the expression pattern of an engrailed protein during the formation of the embryonic (protoconch) and adult shell (teleoconch). We found that in the trochophore-like larva the engrailed protein is expressed in shell-secreting cells at the margin of the protoconch close to the mantle edge. During metamorphosis the growth of the protoconch and expression of the engrailed protein along its margin stop and the teleoconch starts to form. These data suggest a different genetic background regarding protoconch and teleoconch formation in the Scaphopoda and possibly all Conchifera, thus inferring a different evolutionary origin of both organs. The single anlage of the scaphopod protoconch contradicts earlier hypotheses of a monophyletic taxon Diasoma (Scaphopoda + Bivalvia), which has been mainly based on the assumption of a primarily bilobed shell in both taxa. Comparative data on engrailed expression patterns suggest nervous system patterning as the basic function of engrailed in the Bilateria. However, there are several independent gain-of-function events, namely segment compartmentation in the Annelida and Arthropoda, protoconch formation in the Mollusca, skeletogenesis in the Echinodermata, and limb formation in vertebrates. These findings provide further evidence that homologous genes may act in very different pathways of bilaterian body plan formation in various animal phyla.     

Larval and early post-larval shell of three deep-sea Scaphopoda (Mollusca) from the southwest Atlantic

The larval shells of Antalis circumcincta (Watson, 1879) (order Dentaliida), Pertusiconcha callithrix (Dall, 1889) (order Gadilida) and of an undetermined species of uncertain systematic position are described. The material studied comprises mainly samples from deep waters, collected by expeditions along the southeast coast of Brazil. The larval shell of the three taxa matches other types previously described in the literature. Antalis circumcincta and P. callithrix have four regions (protoconch A, protoconch B, teleoconch A, teleoconch B), but differ in dimensions and sculpture from each other, while the undetermined species has three regions (protoconch A, protoconch B, teleoconch B). A morphometric approach combined with a discriminant analysis also indicates that the three taxa are significantly distinct. This study confirms patterns of larval shells at the taxonomic rank of orders but other supraspecific patterns remain uncertain.     

Ultrastructure of the osphradium of Siphonaria grisea (Mollusca, Pulmonata)

By means of scanning and transmissive electron microscopic methods osphradium of Siphonaria grisea has been studied. The osphradium of the animal is presented as a small torulus formed by supporting ciliated cells. Among them bodies of receptory cells are situated; they are of smaller size and decorated with a bundle of microvilli. Central processes of the receptory cells, penetrating through a thick layer of the connective tissue, reach the subepithelial neural trunk. Concentration of the neural cells in the periphery of the trunk is small, and in the central area no chemical synapses are revealed. The data presented demonstrate a primitive structure of the osphradial chemoreceptory organ in Siphonaria.     

Ultrastructure and Phylogenetic Significance of Notaspidean Spermatozoa (Mollusca, Gastropoda, Opisthobranchia)

Spermatozoa of five notaspidean opisthobranchs [Berthellina citrina, Berthella ornata, Pleuro-branchus peroni, Pleurobranchaea maculata, Umbruculum sinicum] were examined using TEM. In all five species, the acrosome (sensu lato) consists of an apical vesicle (the acrosomal vesicle) and acrosomal pedestal. The acrosomal pedestal overlaps the nuclear apex, and in P. peroni (and possibly B. ornata) is periodically banded—-the first reported incidence of this type of substructure in any euthyneuran acrosome. Although sperm nuclei of P. peroni, B. ornata and B. citrina differ in length and also the number of keels present (nucleus 7 μm long with four/five keels present in Pleurobranchus; 17 μm long with one keel in Berthella; 15 μm long with a very weak keel in Berthellina), the basal invagination to which the centriolar derivative, axoneme and coarse fibres are attached is always poorly developed, and very little overlap between nucleus and midpiece occurs. In P. maculata and U. sinicum, the nucleus forms a helical cord around the axoneme and mitochondrial derivative such that it is not possible to recognize exclusively ‘nuclear’ and ‘midpiece’ regions of the spermatozoon. In all notaspideans investigated, (1) the axoneme, coarse fibres and glycogen helix are enclosed by the paracrystalline and matrix components of the mitochondrial derivative and (2) a dense ring structure (attached to the plasma membrane) and glycogen piece are observed. While the glycogen piece is very short (0.85–1.43 μm) with a very degenerate axoneme in B. citrina, B. ornata and P. peroni, this region of the spermatozoan is well developed (30–35 μm long) in U. sinicum and exhibits a fully intact 9 + 2 axoneme. The ‘glycogen piece’(or its presumed homologue) in P. maculata spermatozoa is very short (0.65 μm), devoid of any axonemal remnant and constructed of a hollow, internal cylinder attached to an outer (incomplete) shell, and contains scattered (glycogen) granules. Spermatozoal structure supports a close relationship between the genera Berthellina, Berthella and Pleurobranchus. These three genera have more distant links with Pleurobranchaea, while Umbraculum maintains an isolated, specialized position within the Notaspidea.     

Ultrastructure of spermatozoa in Orthalicidae (Mollusca,Gastropoda, Stylommatophora) and its systematic implications

Sperm morphology of orthalicid gastropods Clessinia pagoda, Spixia tucumanensis, Plagiodontes daedaleus (Odontostominae) and Drymaeus hygrohylaeus, D. poecilus, Bostryx stelzneri (Bulimulinae) are examined and described for the first time using transmission electron microscopy. Spermatozoa show the general characteristic of Pulmonata: an acrosomal vesicle, sperm nucleus helical, mitochondrial derivative forming a continuous sheath with paracrystalline material and coarse fibers associated with axonemal doublets. Features in the acrosomal complex and shape of the nucleus distinguish orthalicid sperms from other stylommatophoran. The acrosomal pedestal is traversed by fine striations in all species examined except in S. tucumanensis. The structure and thickness of the perinuclear sheath with a single or double layer of electron-dense material ensheathing the nuclear apex is characteristic of the group. The presence of a subnuclear ring in Drymaeus, Bostryx and Clessinia species is also reported. A data matrix of eleven species per 34 characters (16 sperm plus 18 anatomical and shell characters) from orthalicids plus other stylommatophoran and systellommatophoran representative species was constructed. Three cladistic analyses (sperm-based, anatomical-based and a combined sperm + anatomical-based) were performed to test the phylogenetic potential of sperm ultrastructure in orthalicid systematics and understand how sperm characters affect the topology and resolution of the obtained trees. Stylommatophora resulted in a monophyletic clade in the sperm-based and in the combined-character analysis. Orthalicidae is monophyletic only in the combined-character cladogram. Within Orthalicidae, Odontostominae is recovered as a monophyletic clade in all analyses, while Bulimulinae is paraphyletic in all trees except in the combined phylogeny. The present study and cladistic analyses performed support the hypothesis that characters on sperm ultrastructure are informative for stylommatophoran systematic and phylogenetic approaches, providing synapomorphies at familiar, subfamiliar and generic level.     

Ultrastructure of multicellular foregut glands in selected Solenogastres (Mollusca)

Foregut glands in Solenogastres are not only of importance in biological processes like feeding and digestion, but multicellular foregut glands also provide valuable characters for taxonomy and systematics. Here, the fine structure of four different types of foregut glands is investigated: the Meioherpia-type ventrolateral foregut gland of Meioherpia atlantica, the Simrothiella-type ventrolateral foregut gland of Simrothiella cf. margaritacea, and the Pararrhopalia-type ventrolateral foregut gland as well as the dorsal gland of Pararrhopalia pruvoti. Thereby, special focus is set on the arrangement of glandular and supporting cells within the glands, and on the characterization of glandular cells according to the size, shape and electron-density of their secretional vesicles. It is shown that the investigated glands are complex organs composed of non-glandular supporting cells and two to five glandular cell types producing discrete secretions.     

The burrowing process of Dentalium (Scaphopoda)

Investigations of the burrowing activity of Dentalium , using cine film and electronic recording techniques, have shown it to penetrate the sand in a series of steps, each termed a "digging cycle". Cycles involve first, pedal dilation, second, retraction followed by extension and probing of the foot. The epipodial lobes are elevated during pedal dilation and form a secure pedal anchor so that at retraction the shell is drawn down over the foot.
A comparison of the burrowing process in the Scaphopoda with that of the Bivalvia indicates that essentially the same mechanisms and sequence of activities are involved, for in both digging consists of the integration of pedal protraction and retraction with the application of shell and pedal anchors. The principal differences, such as the absence in Dentalium of water jets to loosen the sand and high pressures in the pedal haemocoele, are related to the form of their shell. The strength of the pedal anchor was determined and, relative to the weight of Dentalium , is comparable to that of bivalves. In contrast the probing force was relatively weak since the shell anchor of Dentalium , which holds the shell still during probing, is largely limited to its own weight, whereas that attained by the Bivalvia is principally due to the valves being pressed against the substrate by the opening moment of the ligament.     

An ultrastructural analysis of Dentalium vulgare (Mollusca, Scaphopoda) gametes with special reference to early events at fertilization

The ultrastructure of Dentalium gametes, and their fate at fertilization, were investigated. Unexpectedly, fixation carried out in 2% osmium tetroxide, at 4 degrees C, although destroying most cytoplasmic organelles, reveals the presence of bundles of "stress fibers" in the microvilli and the fertilization cone of the eggs. Anti-actin labeling supports the view that the bundles are made of actin. Fertilization, as in other molluscs, does not cause any cortical granule exocytosis, at least during the first 15 min following insemination. Ultrastructure of the unreacted and reacted spermatozoon is presented and a tentative model for the morphological interpretation of the acrosomal reaction is proposed. A reevaluation of cytoplasmic heterogeneity, e.g., the so-called cytoplasmic prelocalization, as observed after the germinal vesicle breakdown is also provided as well as an analysis of concomitant surface changes.     

Molecular phylogeny of Scaphopoda (Mollusca) inferred from 18S rDNA sequences: support for a Scaphopoda–Cephalopoda clade

The phylogenetic relationships of the Scaphopoda, one of the 'lesser' molluscan classes, with the other conchiferan taxa are far from clear. They appear either as the sister-group to the Bivalvia (Diasoma concept) or to a Gastropoda–Cephalopoda clade or to the Cephalopoda alone (helcionellid concept). We compiled a 18S rDNA sequence dataset of 48 molluscan species containing 17 scaphopods to test these hypotheses and to address questions regarding high-level relationships with the Scaphopoda. Both parsimony and maximum likelihood trees show low branch support at the base of the Conchifera, except for the robust clade uniting Scaphopoda and Cephalopoda. This result is corroborated by spectral analysis and likelihood mapping. We also tested alternative topologies which scored significantly worse both in tree length and in likelihood. The 18S rDNA data thus reject the Diasoma in favour of a Scaphopoda–Cephalopoda clade as proposed in the helcionellid concept. When plotted on the molecular tree, the pivotal morphological characters associated with the burrowing life style of the Bivalvia and Scaphopoda, i.e. mantle/shell enclosure of the body and the burrowing foot with true pedal ganglia, appear convergent in these groups. In contrast, the prominent and tilted dorsoventral body axes, multiple cephalic tentacles and a ring-shaped muscle attachment on the shell are potential synapomorphies of Scaphopoda and Cephalopoda. Most of the higher taxa within the Scaphopoda are supported by the molecular data. However, there is no support for the families Dentaliidae and Gadilidae. The basal position of the Fustiariidae within the Dentaliida is confirmed.     

The heart Ultrastructure of lepidopleurus asellus (Spengler) and Tonicella marmorea (Fabricius) (Mollusca: Polyplacophora)

Summary The ultrastructure of the auricles, the ostia, and the ventricle of L. asellus and T. marmorea is described. The heart wall consists of an epicardium, a basement membrane, and an inner loose myocardium. The epicardial cells of the auricle are podocytes. The exposed cell body and the branched processes show pedicles. Ventricular epicardium is flat and simple. The slender, unbranched, mononucleated muscle fibres have a peripheral nucleus located midway along the fibre. Mitochondria are peripherally located, leaving the center to longitudinally running thick and thin myofilaments. Dense bodies and attachment plaques make up the Z-material. Sarcomeres and myofibrils are absent, as are transverse tubules and intercalated disks. The sarcoplasmic reticulum consists of peripheral tubules and subsarcolemmal cisternae, some of which radiate, branch, and run between myofilaments. Couplings are lacking. Ventricular fibres in T. marmorea show nexuses and desmosomes; in L. asellus only nexuses. The muscular ostia are tubular, and muscle fibres resemble those of the ventricle; nexuses are detected in T. marmorea and desmosomes in L. asellus. The only nervous elements observed are some nerve processes, structurally similar to those of other molluscs.Supported by grants from the Norwegian Research Council for Science and Humanities