Larval Tentacles Research Articles

Hox gene expression during development of the phoronid Phoronopsis harmeri

BackgroundPhoronida is a small group of marine worm-like suspension feeders, which together with brachiopods and bryozoans form the clade Lophophorata. Although their development is well studied on the morphological level, data regarding gene expression during this process are scarce and restricted to the analysis of relatively few transcription factors. Here, we present a description of the expression patterns of Hox genes during the embryonic and larval development of the phoronid Phoronopsis harmeri.ResultsWe identified sequences of eight Hox genes in the transcriptome of Ph. harmeri and determined their expression pattern during embryonic and larval development using whole mount in situ hybridization. We found that none of the Hox genes is expressed during embryonic development. Instead their expression is initiated in the later developmental stages, when the larval body is already formed. In the investigated initial larval stages the Hox genes are expressed in the non-collinear manner in the posterior body of the larvae: in the telotroch and the structures that represent rudiments of the adult worm. Additionally, we found that certain head-specific transcription factors are expressed in the oral hood, apical organ, preoral coelom, digestive system and developing larval tentacles, anterior to the Hox-expressing territories.ConclusionsThe lack of Hox gene expression during early development of Ph. harmeri indicates that the larval body develops without positional information from the Hox patterning system. Such phenomenon might be a consequence of the evolutionary intercalation of the larval form into an ancestral life cycle of phoronids. The observed Hox gene expression can also be a consequence of the actinotrocha representing a “head larva”, which is composed of the most anterior body region that is devoid of Hox gene expression. Such interpretation is further supported by the expression of head-specific transcription factors. This implies that the Hox patterning system is used for the positional information of the trunk rudiments and is, therefore, delayed to the later larval stages. We propose that a new body form was intercalated to the phoronid life cycle by precocious development of the anterior structures or by delayed development of the trunk rudiment in the ancestral phoronid larva.

Possible co-option of engrailed during brachiopod and mollusc shell development.

In molluscs, two homeobox genes, engrailed (en) and distal-less (dlx), are transcription factors that are expressed in correlation with shell development. They are expressed in the regions between shell-forming and non-shell-forming cells, likely defining the boundaries of shell-forming fields. Here we investigate the expression of two transcription factors in the brachiopod Lingula anatina We find that en is expressed in larval mantle lobes, whereas dlx is expressed in larval tentacles. We also demonstrate that the embryonic shell marker mantle peroxidase (mpox) is specifically expressed in mantle lobes. Our results suggest that en and mpox are possibly involved in brachiopod embryonic shell development. We discuss the evolutionary developmental origin of lophotrochozoan biomineralization through independent gene co-option.

Organization and metamorphic remodeling of the nervous system in juveniles of Phoronopsis harmeri (Phoronida): insights into evolution of the bilaterian nervous system.

BackgroundMetamorphic remodeling of the nervous system and its organization in juvenile may shed light on early steps of evolution and can be used as an important criterion for establishing the relationships among large groups of animals. The protostomian affiliation of phoronids does not still have certain morphological and embryological proofs. In addition, the relationship of phoronids and other former “lophophorates” is still uncertain. The resolving of these conflicts requires detailed information from poorly investigated members of phoronids, such as Phoronopsis harmeri.ResultsDuring metamorphosis, the juvenile consumes the nerve elements of the larval hood. Two dorsolateral groups of larval perikarya remain and give rise to the dorsal ganglion, which appears as the “commissural brain”. The juvenile inherits the main and minor tentacular nerve rings from the larva. Although the larval tentacles are directly inherited by the juvenile in P. harmeri, the ultrastructure and location of the definitive tentacular neurite bundles change greatly. Innervation of the juvenile lophophore exhibits a regular alternation of the intertentacular and abfrontal neurite bundles. The giant nerve fiber appears at early stage of metamorphosis and passes from the right group of dorsolateral perikarya to the left side of the body.DiscussionThe metamorphic remodeling of the phoronid nervous system occurs in two different ways: with complete or incomplete destruction of organ systems. The morphology of the lophophore seems similar to those of the former members of “Lophophorata”, but its innervation differs greatly. These findings support the separation of bryozoans from Lophophorata and establish a need for new data on the organization of the brachiopod nervous system. The nervous system of the phoronid juvenile is organized as an epidermal nerve plexus but exhibits a nerve center in the anterior portion of the body. The simultaneous presence of both the apical organ and anlage of the cerebral ganglion in phoronids at the larval stage, and the reduction of the apical organ during metamorphosis support the Trochea theory and allow to suggest the presence of two nervous centers in the last common ancestor of the Bilateria. Phoronids retained some plesiomorphic traits and can be regarded as one of the most primitive groups of lophotrochozoans.

Tentacular apparatus ultrastructure in the larva ofBolinopsis infundibulum(Lobata: Ctenophora)

Borisenko, I. and Ereskovsky, A.V. 2011. Tentacular apparatus ultrastructure in the larva of Bolinopsis infundibulum (Lobata: Ctenophora). —Acta Zoologica (Stockholm) 00: 1–10. Most ctenophores have a tentacular apparatus, which plays some role in their feeding. Tentacle structure has been described in adults of only three ctenophore species, but the larval tentacles have remained completely unstudied. We made a light and electron microscopic study of the tentacular apparatus in the larvae of Bolinopsis infundibulum from the White Sea. The tentacular apparatus of B. infundibulum larvae consists of the tentacle proper and the tentacle root. The former contains terminally differentiated cells, while the latter contains stem cells and cells undergoing differentiation. The core of the tentacle is formed by myocytes, and its epidermis contains colloblasts (hunting cells), wall cells, degenerating cask cells, refractive vesicles, and ciliated sensory cells. Stem cells, colloblasts, and cask cells at various stages of differentiation and putative myocytes progenitors were revealed in the tentacle root. Two different populations of the stem cells in the tentacle root give rise to epidermal (colloblasts and cask cells) and mesogleal (myocytes) cell lines. Nervous elements, glandular cells, and basal lamina were not found. Step-by-step differentiation of colloblasts and cask cells is described.

Comparison of the neuromuscular systems among actinotroch larvae: systematic and evolutionary implications

A comparative analysis of the larval and presumptive juvenile neuromuscular systems among actinotroch larvae was performed using confocal laser microscopy with probes for F-actin and serotonin. Currently, there are two main categories of larval nervous systems based on the origin of the nerve fibers that innervate the larval tentacles. Characteristics of the serotonergic cells of the larval apical ganglion and juvenile nervous system have remained relatively conserved, but the structure of the secondary (hood) sense organ and the juvenile tentacles has diversified among species. Differences in larval musculature are mainly associated with differences in hood morphology. The presumptive, juvenile neuromuscular system is either integrated or separated from that of the larva based on the origin of the juvenile tentacles. Among species, the juvenile tentacles are made by remodeling the larval tentacles, developed from a basal tentacular thickening, or developed as a completely separate set in the larva. Differentiation of the neuromuscular structures of the juvenile tentacles is more diverse than their outward morphological characteristics would suggest. Importance of these larval characters is discussed in terms of current problems that exist within phoronid systematics. Evolutionary implications of these morphological characters are discussed among the phoronids, brachiopods, and related bilaterians. Overall, the integration or separation of larval and juvenile neuromuscular characters may yield insights into the evolution of lophotrochozoan body plans.

Structure and metamorphic remodeling of the larval nervous system and musculature of Phoronis pallida (Phoronida).

The structure of the larval nervous system and the musculature of Phoronis pallida were studied, as well as the remodeling of these systems at metamorphosis. The serotonergic portion of the apical ganglion is a U-shaped field of cell bodies that send projections into a central neuropil. The majority of the serotonergic cells are (at least) bipolar sensory cells, and a few are nonsensory cells. Catecholaminergic cell bodies border the apical ganglion. The second (hood) sense organ develops at competence and is composed of bipolar sensory cells that send projections into a secondary neuropil. Musculature of the competent larva includes circular and longitudinal muscle fibers of the body wall, as well as elevators and depressors of the tentacles and hood. The juvenile nervous system and musculature are developed prior to metamorphosis and are integrated with those of the larva. Components of the juvenile nervous system include a diffuse neural net of serotonergic cell bodies and fibers and longitudinal catecholaminergic fibers. The juvenile body wall musculature consists of longitudinal fibers that overlie circular muscle fibers, except in the cincture regions, where this pattern is reversed. Metamorphosis is initiated by the larval neuromuscular system but is completed by the juvenile neuromuscular system. During metamorphosis, the larval nervous system and the musculature undergo cell death, and the larval tentacles and gut are remodeled into the juvenile arrangement. Although the phoronid nervous system has often been described as deuterostome-like, these data show that several cytological aspects of the larval and juvenile neuromuscular systems also have protostome (lophotrochozoan) characteristics.

Ultrastructure and formation of the body cavity lining in Phoronis muelleri (Phoronida, Lophophorata)

Among other characteristics a trimeric coelomic compartmentation consisting of an anterior protocoel, followed by a mesocoel and a posterior metacoel is traditionally believed to substantiate the sister-group relationship between Lophophorata and Deuterostomia, together forming the Radialia. As molecular data cannot support this hypothesis a reanalysis of the coelomic cavities in Phoronida is undertaken, because corresponding coelomic compartmentation is widely accepted to support the Radialia hypothesis. A coelomic cavity can be recognized on the ultrastructural level because its lining is a true epithelium with polarized cells interconnected by apical adherens junctions. This study reveals that neither in larval nor adult Phoronis muelleri (Phoronida) an anterior cavity with such a lining is present. What on the light microscopic level leads to the impression of a cavity inside the larval episphere, actually is an enlarged subepidermal extracellular matrix with an amorphous, presumably gel-like filling, into which several muscle cells are embedded. Larvae, thus, possess only one coelomic cavity, the large trunk coelom of the larva which is adopted in the adult organization. The second coelomic cavity of adult P. muelleri, the lophophore coelom, develops as a double-layer of epithelialized mesodermal cells at the base of the adult tentacle buds and becomes fluid filled during metamorphosis. Like the larval episphere, larval tentacles and most parts of the blastocoel are filled by an amorphous matrix. Reanalysis of the literature and comparison with Brachiopoda and Bryozoa allows the hypothesis that a protocoel is lacking in all Lophophorata, and that merely two unpaired coelomic cavities, one tentacle and one trunk coelom, can be assumed for the ground pattern of this taxon. These results do not provide further evidence for the Radialia hypothesis, but also do not contradict it.

Ultrastructure of the larval tentacle and its skeletal muscle in Xenopus laevis

During premetamorphic development, tadpoles of Xenopus laevis possess a transitory pair of long, slender, mobile tentacles situated at the corners of the mouth. Microscopic examination of the larval tentacle typically reveals three distinct compartments: a central core of cartilage, a laterally situated skeletal muscle, and a nerve supply medially. Along the length of each tentacle, the epidermis is supplied by many unmyelinated nerve fibers, presumably sensory in nature, which terminate as naked axons in close association with the epidermal cells. The striated tentacular muscle, in the proximal region of the lateral compartment, consists of extrafusal muscle fibers of varying size which range in number from 36 to 48 per tentacle (n = 10). Using morphometric criteria, we have classified the skeletal muscle fibers of the larval tentacular muscle into three types: large (30–50 μm), intermediate (20–30 μm), and small (10–20 μm). By electron microscopy, each type displays characteristic sarcomeric banding patterns, sarcotubular and mitochondrial disposition, and motor endplate ultrastructure. Our morphological observations indicate that the tentacles of the Xenopus tadpole are complex mobile facial extensions which may play roles in mechanoreception and/or chemoreception during the waterborne stages of development. Because of its transitory nature, the Xenopus tentacle may be a useful experimental model in future studies of neuromuscular development and subsequent regression in a relatively short period of time.

REVIEW: Lycaenid-ant interactions of the Maculinea type: tracing their historical roots in a comparative framework

Parasitic relationships between lycaenid butterfly larvae and ants are contrasted with the more common, and ancestral, mutualistic butterfly-ant associations. Three types of lycaenid-ant parasitism can be recognized: (i) the Miletinae type (derived from predation on Homoptera); (ii) the Aphnaeini type (derived from obligate and specific mutualism or commensalism); and (iii) the Maculinea type (derived from facultative mutualism, only represented by the genera Maculinea and Lepidochrysops). Parasitic lycaenid-ant interactions are rare (37 confirmed recorded cases worldwide) and are mostly confined to single species, or small species-groups, in larger non-parasitic clades. Only the two genera representing the Maculinea type have undergone substantial radiation. Ant-parasitic lycaenids predominantly occur in highly seasonal habitats with an extended unfavourable dry or cold season. These observations suggest that (a) parasitic interactions are relatively unstable in evolutionary time, and (b) their evolution usually started with caterpillars seeking shelter in ant nests. Ecological and zoogeographical data for species of the Glaucopsychiti (of which Maculinea forms a part) are compiled and used to discuss the evolutionary origin of ant-parasitism in Maculinea butterflies in a comparative framework. These comparisons suggest that (a) Maculinea evolved from facultative unspecific ant-mutualists, (b) the loss of larval tentacle organs is primarily related to their endophytic feeding habits, (c) that Lamiaceae were the hostplants of ancestral Maculinea, and (d) that Maculinea evolved in eastern central Asia. Further phylogenetic and biogeographical analyses are required to quantitatively test these possibilities. © Rapid Science Ltd. 1998

The development of the Merkel cells in the tentacles of Xenopus laevis larvae.

The Merkel cells in the larval tentacles of Xenopus laevis were examined by TEM. Different forms of Merkel cells were found, depending on the age of the larvae or the location in the tentacles. These forms have the appearance of intermediate states between Merkel cells and superficial epidermal cells; thus an epidermal origin for the Merkel cells seems more likely than an immigration from the neural crest. The forms differ in (1) their location in the epidermis, (2) their shape, (3) the number and extension of their desmosomes, (4) the content and distribution of dense-core granules, and (5) the outgrowth of their finger-like processes. Also the relation to a nerve ending is different. By marking Merkel cells with quinacrine, fluorescence spots were observed between the superficial and basal epidermal cells or, in the very tip, within the superficial epidermal cells. These latter spots represent immature Merkel cells, as confirmed by TEM. This indicates a development of Merkel cells from superficial epidermal cells and migration towards the basal layer. Dermal Merkel cells were never observed.

Functional morphology of the larval tentacles of Phragmatopoma californica (Polychaeta: Sabellariidae): composite larval and adult organs of multifunctional significance

Larvae of the sabellariid polychaete Phragmatopoma californica (Fewkes), which were collected in San Diego, California and were competent to metamorphose after 18 to 30 d of development were observed in vivo by videoequipped light microscopy, and the fine structure of the larval tentacles was examined by transmission and scanning electron microscopy. Each tentacle has tufts of at least two types of immotile cilia arranged in dorsolateral and ventrolateral rows, and a ventral groove covered by two types of motile cilia that beat independently of each other. The epidermis is regionalized into glandular, sensory, locomotory, and support cell types and contains four longitudinal bundles of basiepithelial nerve fibers. A layer of connective tissue separates the epidermis and the nerve tracts from obliquely striated muscles that occur within the peritoneum that lines the central coelomic cavities. The peritoneum forms an intact coelomic epithelium that overlies and interdigitates with the muscle cells, with no intervening basal lamina. The muscle cells are considered to be intraperitoneal because they are located above the basal lamina and they lack intercellular junctions with the peritoneal cells. Specialized peritoneal cells form a striated myoepithelial blood vessel that partitions the coelom into medial and lateral cavities. No neuromuscular junctions were found, but both muscular and ciliary movement seem to be under neuronal control. The basiepithelial nerve terminals appear to synapse into the connective tissue layer toward the intraperitoneal muscle. Several similarities in tissue organization are noted between the larval tentacles of P. californica and the tube feet of echinoderms. Observations on the ontogeny, morphology, and behavior of the tentacles suggest that they are multifunctional organs involved in feeding, construction of the juvenile sand tube, locomotion, attachment, and sensory perception during larval and adult lives.

Ultrastructure and behavior of the larva of Phragmatopoma californica (Polychaeta: Sabellariidae): identification of sensory organs potentially involved in substrate selection

The tentacles of the larvae of Phragmatopoma californica (Fewkes) a tubicolous, reef-building polychaete, were examined by video-equipped light microscopy and transmission and scanning electron microscopy. The surface of the tentacles has a unique ciliation pattern, consisting of dorsal tufts of short immotile cilia, ventrolateral tufts of short and long immotile cilia, and ventral motile cilia. Cells bearing immotile cilia are primary sensory cells with long basal processes that form synapses with basiepithelial nerve fibers. The sensory cell cytoplasm is similar to that of nervous tissue, and contains microtubules, neurofilaments, and synaptic vesicles. Sensory cell synapses with basiepithelial nerves appear to be both axodendritic and axoaxonic. The structure of the immotile cilia is compared to that of motile cilia. Unlike motile cilia, immotile cilia are short, rigid, end in a blunt tip and possess and axoneme with typically arranged mictotubules that terminate in an electron-dense end plate. The basal feet of immotile cilia do not anastomose with adjacent basal bodies, and the ciliary membrane is loosely applied to the axoneme and is covered by a surface coat of filamentous material. The use of the larval tentacles during substrate exploration, and the location and ultrastructure of sensory cilia, indicate that they may be involved in the perception of substrateassociated chemical signals and/or mechanical cues of significance in substrate selection.

Of crazy-ants and Curetinae: are Curetis butterflies tended by ants?

The larvae of the lycaenid subfamily Curetinae have never been reported to be associated with ants. Observations on Curetis regula Evans from Brunei are presented which show that this species may be tended by ants both as larvae and adults. The observations are discussed in relation to a recent review on lycaenid/ant associations, u is suggested that the Curetinae will be found to be associated with ants when more species have been reared, on evidence of the larval tentacle organs and apparent ‘pore cupolas’, both of which are ant adaptations. More studies are needed on Curetis biology and larval morphology to resolve the relationships of this enigmatic genus within the Lycaenidae.

The larval development of three species ofMagelona(Polychaeta) from localities near Plymouth

Fertilizations ofMagelona filiformis, M. alleni, andM. mirabiliswere made in the laboratory and the resulting larvae reared through the earliest stages. Later stages of all three species were obtained from the plankton and induced to metamorphose in the laboratory. The long larval tentacles derive from the prototroch and associated tissues. Adult tentacles appear during later pelagic life as thickenings at the bases of the larval tentacles; the latter, together with the long provisional bristles, are discarded at metamorphosis. The early larvae ofM. alleniare considerably larger than equivalent stages of the other two species. The early larvae ofM. mirabilisare peculiar in that at first the larval tentacle of the left side is a mere stump while that of the right side is relatively large; in the other two species both tentacles are of equal length. Larvae of the three species are readily identifiable by characters described in the text.

Neurite complexes with Merkel cells in larval tentacles of Xenopus laevis

Electron microscopy of larval tentacles of Xenopus laevis tadpoles revealed that they are richly innervated structures containing a high concentration of neurite complexes with epidermal Merkel cells. Myelinated sensory nerve fibres supplying each tentacle are abundant. Prior to penetrating the epidermis, they gradually shed their myelin sheaths and Schwann-cell coverings, and subsequently end as naked axon terminals within the epidermis. Many of these intra-epidermal neurites terminate in intimate synaptic contact with granulated Merkel cells. Groups of dense-cored vesicles characterize the Merkel cell, while clusters of clear synaptic vesicles occupy the adjacent sensory nerve terminals. At sites of synaptic contact, the two cells often exhibit an increased thickening of their apposing membranes. Such Merkel cell — sensory neurite complexes occur as isolated units and are profusely scattered throughout the extent of each tentacle. Their ultrastructure is, in most respects, similar to that described previously in specialized regions of other vertebrate and amphibian species. It is suggested that they represent the morphological basis for tactile sensitivity and impart a mechanoreceptive function to the larval tentacle prior to metamorphosis. The larval tentacle in this anuran species may represent a new and convenient research model for future developmental and experimental studies of trophic interactions between sensory nerve fibres and Merkel cells.

Larval development and metamorphosis ofPhoronis psammophila (Phoronida, Tentaculata)

The larval development ofPhoronis psammophila Cori is divided into 6 phases (on the basis of increasing pairs of larval tentacles); furthermore an initial and a ripe phase are distinguished. Specific aspects of the development are described: Formation and structure of larval tentacles; anlage of adult tentacles as a thickening in the larval tentacle base; late development of the metasome (larva with 4–6 tentacles); formation of the metasome pouch in the larva with 8 tentacles; enlargement of the apical plate; differentiation of the gut; differentiation of larval nephridia; formation of pigment particles in the larva with 6 tentacles (storage function of pigments and its significance for larval identification); different types of discoflagella in various regions of the body. The larval development shows the following tendencies: Improvement of locomotion; intensification of food filtration; anlage of adult organs in the larva leading to a shortening of metamorphosis duration. The larva ofP. psammophila is compared with those ofP. pallida, P. hippocrepia, andP. vancouverensis. Earlier larval determinations ofP. psammophila (e.g.Actinotrocha sabatieri, A. hatschekii) are shown to have been mistakes. Termination of the postembryonic phase (metamorphosis) can be induced experimentally by bacteria and also by cations. Pure or mixed bacteria cultures must be present at the beginning exponential growth phase. The bacteria density required is 20–94×106 bact.ml−1 for pure cultures and on the average 28×106 bact. ml−1 for mixed cultures. Metamorphosis initiation by cations can be induced with CsCl (0.06 M) and RbCl (0.035 M). Metamorphosis ofP. psammophila occurs in 6 phases: larva, ready for metamorphosis; larva, activated by bacteria or ions; evagination of the metasome diverticle, dislocation of gut; losing and swallowing of episphaere and larval tentacles; formation of the youngP. psammophila. All developmental phases are described and compared with those ofP. muelleri; imperfect metamorphosis is characterized and the youngP. psammophila compared with older stages and the adult Phoronis.

Scanning electron microscopic observations of the larval development of the reef-building polychaete Phragmatopoma lapidosa

The scanning electron microscope (SEM) was used in a study of the larval development of the reef-building sabellariid polychaete Phragmatopoma lapidosa. Mature sperm are modified from the 'primitive' polychaete plan by possessing a tapered and bent acrosome. The outer surface of the irregularly shaped, freshly spawned egg envelope is granular in appearance. SEM observations of various stages in development from the early trochophore through larval metamorphosis and the early juvenile stages suggest that the egg envelope serves as the cuticle through the trochophore stage but is then replaced by another cuticle penetrated by microvilli. SEM has revealed the presence of 'sensory tufts' on the dorsal surface of the larval tentacles which may play a role in larval substrate selection.

Developmental studies of the post-settling stages of Sabellaria vulgaris (Polychaeta: Sabellariidae)

A laboratory study on the development of Sabellaria vulgaris Verrill, 1873 was conducted to detail such heretofore undescribed post-settling events as the formation of opercular paleae. Culture techniques for the rearing of sabellariid larvae are described. Larval settling behavior is described and the effects of various culturing methods on larval settlement are discussed. Larvae metamorphosed in 19 to 30 days at 21° to 23°C. Metamorphosis involved a loss of provisional setae, anterior rotation of larval tentacles and opercular cirri and a reduction of the episphere. Following settlement, larvae built tubes, lost most of their pigmentation and ciliary bands and added new abdominal segments. Formation of the opercular crown involved the progressive secretion and loss of outer, middle and inner paleae which migrated into the crown after secretion within setigerous sacs. A detailed description of their formation and a discussion of their systematic significance are presented.

A histochemical-physiological correlation of frog skeletal muscle fibers.

A histochemical-physiological correlation of frog skeletal muscle fibers.

Phoronidea from Brazil

Phoronidea from Brazil