The attachment complex of brachiolaria larvae of the sea star <Emphasis Type="Italic">Asterias rubens</Emphasis> (Echinodermata): an ultrastructural and immunocytochemical study

The attachment complex of brachiolaria larvae of the asteroid Asterias rubens comprises three brachiolar arms and an adhesive disc located on the preoral lobe. The former are used in temporary attachment and sensory testing of the substratum, whereas the latter is used for permanent fixation to the substratum at the onset of metamorphosis. Brachiolar arms are hollow structures consisting of an extensible stem tipped by a crown of dome-like ciliated papillae. The papilla epidermis is composed of secretory cells (type A, B and C cells), non-secretory ciliated cells, neurosecretory-like cells and support cells. Type A and B secretory cells fill a large part of the papilla epidermis and are always closely associated. They presumably form a duo-gland adhesive system in which type A and B cells are respectively adhesive and de-adhesive in function. The adhesive disc is an epidermal structure mainly composed of secretory cells and support cells. Secretory cells produce the cement, which anchor the metamorphic larva to the substratum until the podia are developed. The relatedness between the composition of the adhesive material in the brachiolaria attachment complex and in the podia of adults was investigated by immunocytochemistry using antibodies raised against podial adhesive secretions of A. rubens. Type A secretory cells were the only immunolabelled cells indicating that their temporary adhesive shares common epitopes with the one of podia. The attachment pattern displayed by the individuals of A. rubens during the perimetamorphic period—temporary, permanent, temporary—is unique among marine non-vertebrate Metazoa.     

Structure of the organs of attachment of brachiolaria larvae of Stichaster australis (Verrill) and Coscinasterias calamaria (Gray) (Echinodermata: Asteroidea)

The structure of the brachiolar arms and adhesive disk of the brachiolaria larvae of Stichaster australis (Verrill) and Coscinasterias calamaria (Gray) was determined from light microscopy and from scanning and transmission electron microscopy. The structure of these organs was very similar in both species.The brachiolar arms are comprised of a stem region terminating in a crown of adhesive papillae which are made up of a variety of secretory cell types. Principal among these are elongated cells producing very electron-dense secretory particles, which are released at the free cell surface attached to cilia. Secretory particles appear to be important in temporary attachment of the brachiolar arms to the substratum. Ciliary sense cells, possibly used in the recognition of specific substrata are located at the tip of adhesive papillae.The adhesive disk is comprised of large cells packed with secretory droplets and elongated intracellular fibres. In the attached adhesive disk, secretory droplets are lost, having formed the cement that attaches the disk to the substratum. It appears that adhesive papillae lateral to the adhesive disk hold the disk in position close to the substratum during secretion and hardening of the cement. The intracellular fibres are the principal anchoring structures running from microvilli (locked into the attachment cement) on the surface of the disk to the underlying connective tissue of the attachment stalk.     

Adhesive Papillae of Phallusia mamillata Larvae: Morphology and Innervation

The swimming larvae of most solitary ascidians belonging to the Ascidiidae family bear three anterior, simple conic adhesive papillae. They secrete adhesive substances that are used to effect transitory settlement at the beginning of the metamorphosis.The adhesive papillae of newly hatched Phallusia mamillata larvae examined by the SEM are covered by the tunic. When the larvae are about to settle, the tunic becomes fenestrated over the central part of the papilla and bulb-ended microvilli protrude through the holes. These papillae have two types of elongated cells: many peripheral cells and few larger central cells with microvilli and bundles of microtubules oriented along the major axis of the cells.We have done immunofluorescence experiments with an anti-beta-tubulin monoclonal antibody (clone 2-28-33) reacting with axonal microtubules. Only the central cells of the papillae were stained and the axons appeared to arise from the proximal ends of these cells. These axons form a long nerve that reaches the brain vesicle. Branches of the same nerve appear to connect to the basal ends of the peripheral cells. By confocal laser microscopy we were able to follow the course of the papillary nerve. The two nerves connecting the dorsal papillae fuse together into a single nerve that runs posteriorly. The nerve connecting the ventral papilla runs posteriorly for a long tract before fusing with the nerve of the dorsal papillae just near the brain.The reported observations raise the hypothesis that the central cells of the adhesive papillae might be primary sensory neurons and that they may have chemosensory function.     

Adaptations to benthic development: functional morphology of the attachment complex of the brachiolaria larva in the sea star Asterina gibbosa

The asteroid Asterina gibbosa lives all its life in close relation to the sea bottom. Indeed, this sea star possesses an entirely benthic, lecithotrophic development. The embryos adhere to the substratum due to particular properties of their jelly coat, and hatching occurs directly at the brachiolaria stage. Brachiolariae have a hypertrophied, bilobed attachment complex comprising two asymmetrical brachiolar arms and a central adhesive disc. This study aims at describing the ultrastructure of the attachment complex and possible adaptations, at the cellular level, to benthic development. Immediately after hatching, early brachiolariae attach by the arms. All along the anterior side of each arm, the epidermis encloses several cell types, such as secretory cells of two types (A and B), support cells, and sensory cells. Like their equivalents in planktotrophic larvae, type A and B secretory cells are presumably involved in a duo-glandular system in which the former are adhesive and the latter de-adhesive in function. Unlike what is observed in planktotrophic larvae, the sensory cells are unspecialized and presumably not involved in substratum testing. During the larval period, the brachiolar arms progressively increase in size and the adhesive disc becomes more prominent. At the onset of metamorphosis, brachiolariae cement themselves strongly to the substratum with the adhesive disc. The disc contains two main cell types, support cells and secretory cells, the latter being responsible for the cement release. During this metamorphosis, the brachiolar arms regress while post-metamorphic structures grow considerably, especially the tube feet, which take over the role of attachment to the substratum. The end of this period corresponds to the complete regression of the external larval structures, which also coincides with the opening of the mouth. This sequence of stages, each possessing its own adhesive strategy, is common to all asteroid species having a benthic development. In A. gibbosa, morphological adaptations to this mode of development include the hypertrophic growth of the attachment complex, its bilobed shape forming an almost completely adhesive sole, and the regression of the sensory equipment.     

Chronology of Development in the Starfish <Emphasis Type="Italic">Asterina pectinifera</Emphasis> from Vostok Bay,Sea of Japan

The development of the starfish Asterina (= Patiria) pectinifera (Muller et Troschel) from fertilization to metamorphosis took 27–28 days at 22°C and a salinity of 33–33.4‰. The embryonic development was completed by the release of swimming ciliary blastula from egg envelopes 13 h after fertilization. The larvae passed into the stage of gastrula and reached the stage of dipleurula in 35 h and the stage of bipinnaria in 3.5 days. At the stage of brachiolaria, by the 12th day of development, two lateral brachioles and one medioventral brachiole with papillae developed in the larvae. The attachment disk and the primordia of five radial canals of the juvenile starfish became visible by the 15th and 18th days, respectively. By the 24th–25th day, a differentiated primordium of a juvenile starfish had developed in the brachiolaria. The size of the larvae prior to settlement was 1765.4 ± 51.5 µm. Metamorphosis was completed one day after settlement.Original Russian Text Copyright ¢ 2005 by Biologiya Morya, Kashenko.     

Evaluation of the attachment strength of individuals of Asterina gibbosa (Asteroidea, Echinodermata) during the perimetamorphic period

A turbulent channel flow apparatus was used to determine the adhesion strength of the three perimetamorphic stages of the asteroid Asterina gibbosa, i.e. the brachiolaria larvae, the metamorphic individuals and the juveniles. The mean critical wall shear stresses (wall shear stress required to dislodge 50% of the attached individuals) necessary to detach larvae attached by the brachiolar arms (1.2 Pa) and juveniles attached by the tube feet (7.1 Pa) were one order of magnitude lower than the stress required to dislodge metamorphic individuals attached by the adhesive disc (41 Pa). This variability in adhesion strength reflects differences in the functioning of the adhesive organs for these different life stages of sea stars. Brachiolar arms and tube feet function as temporary adhesion organs, allowing repetitive cycles of attachment to and detachment from the substratum, whereas the adhesive disc is used only once, at the onset of metamorphosis, and is responsible for the strong attachment of the metamorphic individual, which can be described as permanent adhesion. The results confirm that the turbulent water channel apparatus is a powerful tool to investigate the adhesion mechanisms of minute organisms.     

ENZYMATIC ACTIVITIES RELATING TO THE RESPIRATION AND THE ONSET OF METAMORPHOSIS OF THE ASCIDIAN TADPOLE

The oxygen consumption of a single ascidian larva was measured. After hatching the consumption increases gradually. During the period of tail resorption it also increases gradually, but after the completion of tail resorption the consumption decreases conspicuously.
With the development of the larva after hatching, the activities of cytochrome oxidase and succinic dehydrogenase and of Janus green-reduction become detectable in the adhesive papillae, the proximal region of the tail, and the tail muscle. After the completion of tail resorption, these activities become indistinct.
These tissues underwent most profound morphological changes at the onset of metamorphosis. Soon after hatching, Janus green has no effect to induce metamorphosis. In larvae 4 hr after hatching, the shrinkage of adhesive papillae can be induced by Janus green-treatment. In 12 hr larvae, both the shrinkage of adhesive papillae and the tail resorption can be induced by Janus green. The enhancement of respiratory activities in the larvae after hatching may be related to the changes in the adhesive papillae and later to changes in the proximal region of the tail. Only when both of these changes occur can metamorphosis be induced.     

Evaluation of the attachment strength of individuals of Asterina gibbosa (Asteroidea,Echinodermata) during the perimetamorphic period

Abstract

A turbulent channel flow apparatus was used to determine the adhesion strength of the three perimetamorphic stages of the asteroid Asterina gibbosa, i.e. the brachiolaria larvae, the metamorphic individuals and the juveniles. The mean critical wall shear stresses (wall shear stress required to dislodge 50% of the attached individuals) necessary to detach larvae attached by the brachiolar arms (1.2 Pa) and juveniles attached by the tube feet (7.1 Pa) were one order of magnitude lower than the stress required to dislodge metamorphic individuals attached by the adhesive disc (41 Pa). This variability in adhesion strength reflects differences in the functioning of the adhesive organs for these different life stages of sea stars. Brachiolar arms and tube feet function as temporary adhesion organs, allowing repetitive cycles of attachment to and detachment from the substratum, whereas the adhesive disc is used only once, at the onset of metamorphosis, and is responsible for the strong attachment of the metamorphic individual, which can be described as permanent adhesion. The results confirm that the turbulent water channel apparatus is a powerful tool to investigate the adhesion mechanisms of minute organisms.     

Reproduction and larval morphology of broadcasting and viviparous species in the Cryptasterina species complex

The Cryptasterina group of asterinid sea stars in Australasia comprises cryptic species with derived life histories. C. pentagona and C. hystera have planktonic and intragonadal larvae, respectively. C. pentagona has the gonochoric, free-spawning mode of reproduction with a planktonic lecithotrophic brachiolaria larva. C. hystera is hermaphroditic with an intragonadal lecithotrophic brachiolaria, and the juveniles emerge through the gonopore. Both species have large lipid-rich buoyant eggs and well-developed brachiolariae. Early juveniles are sustained by maternal nutrients for several weeks while the digestive tract develops. C. hystera was reared in vitro through metamorphosis. Its brachiolariae exhibited the benthic exploration and settlement behavior typical of planktonic larvae, and they attached to the substratum with their brachiolar complex. These behaviors are unlikely to be used in the intragonadal environment. The presence of a buoyant egg and functional brachiolaria larva would not be expected in an intragonadal brooder and indicate the potential for life-history reversal to a planktonic existence. Life-history traits of species in the Cryptasterina group are compared with those of other asterinids in the genus Patiriella with viviparous development. Modifications of life-history traits and pathways associated with evolution of viviparity in the Asterinidae are assessed, and the presence of convergent adaptations and clade-specific features associated with this unusual mode of parental care are examined.     

A gastric-brooding asteroid, Smilasterias multipara

The gastric-brooding asterinid sea star, Smilasterias multipara, broods from late August to early November in the shallow sublittoral zone of southeastern Australia. We observed males and females spawning in the laboratory. They shed gametes through gonopores on the sides of the arms. The eggs were orange, about 1.0 mm in diameter, and heavier than seawater. They were externally fertilized by sperm, and placed into the stomach of the female by the tube feet. Twenty-four hours after fertilization, the first cleavage occurred. Cleavage was equal, total, and radial. Development via a non-feeding lecithotrophic brachiolaria was direct, there being no planktrotrophic bipinnaria or brachiolaria larva. Embryos developed, through wrinkled blastula and gastrula stages, into brachiolariae with arms. All of the surfaces of the brachiolaria were covered by cilia. At metamorphosis, a starfish rudiment appeared on the posterior portion of the larval body, while the anterior portion of the larval body was absorbed. Two months after fertilization, metamorphosis was complete. After metamorphosis, juveniles in the stomach grew six pairs of tube feet in each arm. Juveniles, 3 mm in diameter, emerged from the mouth of the mother in early November. Developmental evidence suggests that this asteroid has evolved mechanisms for the protection of larvae and juveniles from gastric digestion.     

Dopamine and serotonin modulate the onset of metamorphosis in the ascidian Phallusia mammillata

Neurotransmitters play an important role in larval metamorphosis in different groups of marine invertebrates. In this work, the role of dopamine and serotonin during metamorphosis of the ascidian Phallusia mammillata larvae was examined. By immunofluorescence experiments, dopamine was localized in some neurons of the central nervous system and in the adhesive papillae of the larvae. Dopamine and serotonin signaling was inhibited by means of antagonists of these neurotransmitters receptors (R(+)-SCH-23390, a D(1) antagonist; clozapine, a D(4) antagonist; WAY-100635, a 5-HT(1A) antagonist) and by sequestering the neurotransmitters with specific antibodies. Moreover, dopamine synthesis was inhibited by exposing 2-cell embryos to alpha-methyl-l-tyrosine. Dopamine depletion, obtained by these different approaches, caused early metamorphosis, while serotonin depletion delayed the onset of metamorphosis. The opposite effects were obtained using agonists of the neurotransmitters: lisuride, a D(2) agonist, inhibited metamorphosis, while DOI hydrochloride and 8-OH-DPAT HBr, two serotonin agonists, promoted it. So, it is possible to suppose that dopamine signaling delayed metamorphosis while serotonin signaling triggers it. We propose a mechanism by which these neurotransmitters may modulate the timing of metamorphosis in larvae.     

Localization of hatching enzyme in embryos and larvae of the sea-squirt Ciona intestinalis

Summary

A polyclonal antibody raised against the hatching enzyme of Ciona intestinalis (D'Aniello et al., 1997) was used on larvae of different ages in whole mount immunofluorescence experiments in order to localize the cells secreting the enzyme. After staining with FITC-conjugated second antibody, the larvae were observed by confocal microscopy. Larvae just before hatching (9–10 hours after fertilization) showed the presence of the enzyme in the peripheral cells of the adhesive papillae. The newly hatched larvae showed fluorescence also in the epidermal cells of the tip of the tail. Higher magnification confocal images of the papillae revealed bright fluorescence both in peripheral cells of the papillae and in the cavity between the tunic and the apex of the papillae (hyaline cap).

The swimming larvae maintain the fluorescence in the peripheral cells and in the hyaline cap for some hours until the beginning of metamorphosis, whereas the fluorescence of the tip of the tail disappears.

Following application of the antibody to Phallusia mamillata, the peripheral cells of the papillae of the newly hatched larvae were fluorescent and a bright fluorescence was also present between the two layers of the tunic above the papillae and the anterior part of the cephalenteron. We never observed fluorescence in the cells of the epidermis of the tail.

Retinoic acid (RA) treatment has been used to confirm the localization on the papillae of the cells secreting the hatching enzyme. The larvae of Ciona intestinalis were able to hatch because the cells of the tip of the tail positively reacted to immunofluorescence stain with anti-hatching enzyme antibody. On the contrary Phallusia tnamillata larvae failed to hatch and did not show anti-hatching enzyme reaction in the tail.     

Ascidian neural crest-like cells: phylogenetic distribution, relationship to larval complexity, and pigment cell fate

Migratory neural crest-like cells, which express the cell surface antigen HNK-1 and develop into pigment cells, have recently been identified in the ascidian Ecteinascidia turbinata. Here we use HNK-1 expression as a marker to determine whether neural crest-like cells are responsible for pigment development in diverse ascidian species. We surveyed HNK-1 expression and tyrosinase activity in 12 ascidian species, including those with different adult organizations, developmental modes, and larval sizes and complexities. We observed HNK-1 positive cells in every species, although the timing of HNK-1 expression varied according to the extent of larval complexity. HNK-1 expression was initiated during the late tailbud stage in species in which adult features are formed precociously in large complex larvae. In contrast, HNK-1 positive cells did not appear until the swimming tadpole or juvenile stage in species with small simple larvae in which most adult features appear after metamorphosis. Double labeling experiments indicated that HNK-1 and tyrosinase are expressed in the same subset of pigment-forming mesenchymal cells in species with complex or simple larvae. In addition, the absence of HNK-1 and tyrosinase expression in albino morphs of the colonial ascidian Botryllus schlosseri suggested that the major fate of neural crest-like cells is to become pigment cells. The results suggest that ascidian neural crest-like cells and vertebrate neural crest cells had a common origin during chordate evolution and that their primitive function was to generate body pigmentation.     

Hemps, a novel EGF-like protein, plays a central role in ascidian metamorphosis

All chordates share several characteristic features including a dorsal hollow neural tube, a notochord, a pharynx and an endostyle. Unlike other chordate taxa, ascidians have a biphasic life-history with two distinct body plans. During metamorphosis, the larval nerve cord and notochord degenerate and the pharyngeal gill slits and endostyle form. While ascidians, like other marine invertebrates, metamorphose in response to specific environmental cues, it remains unclear how these cues trigger metamorphosis. We have identified a novel gene (Hemps) which encodes a protein with a putative secretion signal sequence and four epidermal growth factor (EGF)-like repeats which is a key regulator of metamorphosis in the ascidian Herdmania curvata. Expression of Hemps increases markedly when the swimming tadpole larva becomes competent to undergo metamorphosis and then during the first 24 hours of metamorphosis. The Hemps protein is localised to the larval papillae and anterior epidermis of the larva in the region known to be required for metamorphosis. When the larva contacts an inductive cue the protein is released, spreading posteriorly and into the tunic as metamorphosis progresses. Metamorphosis is blocked by incubating larvae in anti-Hemps antibodies prior to the addition of the cue. Addition of recombinant Hemps protein to competent larvae induces metamorphosis in a concentration-dependent manner. A subgroup of genes are specifically induced during this process. These results demonstrate that the Hemps protein is a key regulator of ascidian metamorphosis and is distinct from previously described inducers of this process in terrestrial arthropods and aquatic vertebrates.     

Immunohistochemical analysis of adhesive papillae of Clavelina lepadiformis (Müller, 1776) and Clavelina phlegraea (Salfi, 1929) (Tunicata,Ascidiacea)

Almost all ascidian larvae bear three mucus secreting and sensory organs, the adhesive papillae, at the anterior end of the trunk, which play an important role during the settlement phase. The morphology and the cellular composition of these organs varies greatly in the different species. The larvae of the Clavelina genus bear simple bulbous papillae, which are considered to have only a secretory function. We analysed the adhesive papillae of two species belonging to this genus, C. lepadiformis and C. phlegraea, by histological sections and by immunolocalisation of β-tubulin and serotonin, in order to better clarify the cellular composition of these organs. We demonstrated that they contain at least two types of neurons: central neurons, bearing microvilli, and peripheral ciliated neurons. Peripheral neurons of C. lepadiformis contain serotonin. We suggest that these two neurons play different roles during settlement: the central ones may be chemo- or mechanoreceptors that sense the substratum, and the peripheral ones may be involved in the mechanism that triggers metamorphosis.Key words: Settlement, neurotransmitter, serotonin, β-tubulin, papillary nerves, metamorphosis.Ascidians (phylum Chordata; subphylum Tunicata) are sessile filter-feeding organisms that can be found in all benthic marine environments and develop through a swimming tadpole larva. Larvae of colonial ascidians have a short planktonic life that can vary from minutes to hours (Burighel and Cloney, 1997). Prior to metamorphosis, the larva attaches to the substratum by means of peculiar organs of ectodermic origin, located in the anterior region of the trunk. These organs, known as adhesive papillae, secrete sticky substances and effect primary adhesion of the larva to the substrate (Cloney, 1977). They have an important role in the initiation of settlement and metamorphosis and there is evidence that, at least in some species, they participate in substrate selection (Torrence and Cloney, 1983; Svane and Young, 1989; Groppelli et al., 2003). In many species, they are organised in a triangular field, whereas in others they are aligned along the mid-sagittal plane of the trunk. Adhesive papillae have been classified into two types: eversible papillae, typical of some colonial species, composed by several cell types and rapidly changing shape as they touch the substrate, and non-eversible papillae, typical of solitary species, which do not change shape after settlement (Burighel and Cloney, 1997).With few exceptions, all adhesive papillae are formed by elongated secretory and sensory cells, which are recognised as primary neurons (Cloney, 1977, 1979). It has been proposed that sensory cells may detect the chemical and physical characteristics of the substratum at potential sites for settlement and metamorphosis (Young and Braithwaite, 1980; Groppelli et al., 2003).The presence of primary neurons in the papillae has been reported in the larvae of several species such as Distaplia occidentalis, Diplosoma macdonaldi, Phallusia mammillata, Ciona intestinalis and Ascidia malaca (Cloney, 1977;Torrence and Cloney, 1983; Sotgia et al., 1998; Takamura, 1998; Gianguzza et al., 1999).These neurons have axons that join together to form the papillary nerves that enter the central nervous system at the level of the sensory vesicle (Imai and Meinertzhagen, 2007).Recently, different neurotransmitters have been localised in the sensory neurons of the papillae of different species. The presence of GABAergic neurons has been reported in the papillae of Ciona savygni (Brown et al., 2005) and of Ciona intestinalis (Zega et al., 2008), while serotonergic neurons have been localised in the papillae of Phallusia mammillata (Pennati et al., 2001) and Botrylloides leachi (Pennati et al., 2007). Moreover, it has been demonstrated that serotonin plays a role in the mechanism triggering metamorphosis in ascidians (Zega et al., 2005).After attachment, all papillae retract to draw the larva closer to the substratum. In the colonial ascidian Distaplia occidentalis, the process of retraction is reversibly inhibited by cytochalasin B, suggesting that microfilaments are involved in this process (Cloney, 1979).Clavelina lepadiformis is a colonial species, whose larvae bear non-eversible simple bulbous papillae.They have been described as being formed only by columnar glandular cells, whose secreted material is responsible for the sticky properties of the organ. These papillae do not contain sensory cells and were considered the simplest among those studied (Turon, 1991).In this work, the morphology of the adhesive papillae of C. lepadiformis and C. phlegraea was further investigated by histological analysis and immunolabelling techniques in order to clarify the actual cellular composition and function of these organs.     

Comparative analysis of thyroxine distribution in ascidian larvae

The ascidian endostyle is a mucus-secreting pharyngeal organ, it has iodine-concentrating activity and the biosynthesis of thyroid hormones has been well documented. According to our recent findings, ascidians possess thyroid hormones, which are localized in mesenchymal cells. We have studied the presence and localization of l-thyroxine (T₄) in Ascidia malaca (Traustedt), Ascidiella aspersa (Müller), Phallusia mamillata (Cuvier) and Ciona intestinalis (Linnaeus) larvae and its involvement in metamorphosis. In vivo treatment of swimming larvae with exogenous T₄ and thiourea (a thyroid hormone synthesis inhibitor), demonstrate the presence of T4 during larval development. These results were confirmed by in vitro experiments utilizing dot blotting, radioimmunoassay and immunoperoxidase staining. The hormone was localized in mesenchymal cells of all four ascidians, spread out in the body cavity, under the adhesive papillae and around the intestine. The presence of TH in mesenchymal cells could be related to blood cells, musculature and heart tissue differentiation. The results suggest that this hormone could be involved in the control of metamorphosis.     

Morphogenesis and phenotypic divergence in two developmental morphs of Streblospio benedicti (Annelida,Spionidae)

Abstract. The morphology of marine invertebrate larvae is strongly correlated with egg size and larval feeding mode. Planktotrophic larvae typically have suites of morphological traits that support a planktonic, feeding life style, while lecithotrophic larvae often have larger, yolkier bodies, and in some cases, a reduced expression of larval traits. Poecilogonous species provide interesting cases for the analysis of early morphogenesis, as two morphs of larvae are produced by a single species. We compared morphogenesis in planktotrophic and lecithotrophic morphs of the poecilogonous annelid Streblospio benedicti from the trochophore stage through metamorphosis, using observations of individuals that were observed alive, with scanning electron microscopy, or in serial sections. Offspring of alternate developmental morphs of this species are well known to have divergent morphologies in terms of size, yolk content, and the presence of larval bristles. We found that some phenotypic differences between morphs occur as traits that are present in only one morph (e.g., larval bristles, bacillary cells on the prostomium and pygidium), but that much of the phenotypic divergence is based on heterochronic changes in the differentiation of shared traits (e.g., gut and coelom). Tissue and organ development are compared in both morphs in terms of their structure and ontogenetic change throughout early development and metamorphosis.     

Larval adhesive organs and metamorphosis in ascidians

Summary The larva of Distaplia occidentalis bears three cup-shaped adhesive papillae, each with a prominent axial protrusion. At the onset of metamorphosis these organs rapidly evert through fenestrations in the cuticular layers of tunic exposing hyaline caps of adhesive. Additional adhesive material is secreted from collocytes during eversion. The stickiness of the papillae facilitates attachment to a variety of substrates.Each papilla is composed of more than 900 cells; six different types were identified. The wall of the cup contains about 260 myoepithelial cells with long attenuated processes. These extend from the rim of the cup to the base in the parietal (inner) layer. The apices of the myoepithelial cells are held in place by 11 pairs of specialized anchor cells bearing long bulbous microvilli. When the myoepithelial cells contract they force the axial protrusion forward and transform the papilla into a hyperboloidal configuration. The papilla is innervated by small motor fibers, but sensory fibers were not detected. The adhesive papillae of Distaplia are discussed in relationship to nine other recognizable types of papillae in the ascidians.     

Characterization of the lecithotrophic larval development of the temperate New Zealand asterinid Stegnaster inflatus

In the family Asterinidae, development through a planktonic lecithotrophic brachiolaria larva is common and has evolved independently several times. Here, we describe the lecithotrophic development of the asterinid Stegnaster inflatus, a species endemic to New Zealand. Early development through the blastula and gastrula stages is short, with hatching at the brachiolaria stage occurring within 48 hr. After hatching, larvae are negatively buoyant, and without aeration remain near the bottom of the culture containers. The settled benthic juvenile stage was reached in ~2 weeks. The brachiolaria of S. inflatus shares common characteristics with the planktonic brachiolaria of other asterinids in that the brachiolar attachment apparatus comprises three brachia and a central adhesive disc, although the latter is thin and appears to be reduced. Mortensen (1925, Videns kabelige Meddelelser fra Dansk naturhistorisk Forening i København, 79 (15), 261‐420) had hypothesized that individuals of S. inflatus might brood within the “cave” formed in the interambulacral space between the arms. We found no evidence for brooding, but hypothesize that S. inflatus may have demersal development, on or near the bottom, which has implications for larval dispersal and population structure.