Comparative SEM Studies of Lattice Organs: Putative Sensory Structures on the Carapace of Larvae from Ascothoracida and Cirripedia (Crustacea Maxillopoda Thecostraca)

Putative sensory structures, called lattice organs, were studied with scanning electron microscopy from ascothoracid or cypris larvae representing a wide range of families of the Ascothoracida and Cirripedia. These organs, situated dorsally on the carapace were, with few exceptions, always found in two anterior and three posterior pairs. The lattice organ morphology displayed by the Ascothoracida, a seta–like structure with a terminal pore, is believed to be the most plesiomorphic condition. Within the Cirripedia lattice organ morphology varied from types resembling the Ascothoracida in the Acrothoracica and the lepadomorph Capitulum mitella, to an elongate pore field with a larger terminal pore in most Thoracica and Rhizocephala. Akentrogonid Rhizocephala seem to display the most apomorphic condition. While lattice organ morphology was generally constant at the family level, cases were seen where closely related species such as Chthamalus stellatusand Chthamalus montaguishowed minor, but clear cut differences. Lattice organs in 2 + 3 pairs are argued to represent a synapomorphy for the Ascothoracida and the Cirripedia. The results confirm that the cyprid morphology at the ultrastructural level will prove to be of high value in estimating phylogeny within the Cirripedia.     

The Chemoreceptive Lattice Organs in Cypris Larvae Develop from Naupliar Setae (Thecostraca: Cirripedia, Ascothoracida and Facetotecta)

Lattice organs are peculiar chemoreceptors found only in the Crustacea Thecostraca (Facetotecta, Ascothoracida, Cirripedia). In these taxa, five pairs occur in the head shield (carapace) of the terminal larval instar (y-cyprid, ascothoracid larva, cyprid), which is the settlement stage. Lattice organs represent an autapomorphy for the Thecostraca but their evolutionary origin and possible homologues in other Crustacea remain obscure. We have used scanning electron microscopy to describe the setation pattern of the head shield in late nauplii of one species of Ascothoracida, one species of Facetotecta and several species of the Cirripedia Thoracica, Acrothoracica, and Rhizocephala. The naupliar head shield always carries two pairs setae situated anteriorly near the midline. Each of these setae carry a single pore, and positional, structural and ontogenetic evidence show that these setae are homologous in all the examined species and that they represent precursors of the two anterior pairs of lattice organs of the succeeding larval stage, viz., the ascothoracid larva (Ascothoracida), y-cyprid (Facetotecta), and cyprid (Cirripedia). This leads us to infer that lattice organs are among the most highly modified sensilla in all Crustacea and they have in most cases lost all external resemblance to a seta. The nauplii of the Rhizocephala carry an additional three pairs of setae situated more posteriorly on the head shield and they could be precursors of the three posterior pairs of lattice organs. All other species examined lack these posterior setae, except the Facetotecta which have one posteriorly situated pair.     

TEM studies on the lattice organs of cirripede cypris larvae (Crustacea, Thecostraca, Cirripedia)

 Lattice organs consist of five pairs of sensory organs situated on the dorsal carapace in cypris larvae of the Crustacea Cirripedia. The lattice organs in cypris larvae of Trypetesa lampas (Acrothoracica) and Peltogaster paguri (Rhizocephala) represent the two main types found in cirripedes, but only minor differences exist at the TEM level. Each lattice organ is innervated by two bipolar, primary receptor cells. The inner dendritic segment of each receptor cell carries two outer dendritic segments. The outer dendritic segments contain modified cilia with a short ciliary segment (9×2+0 structure). Two sheath cells envelop the dendrite except for the distal ends of the outer dendritic segments. This distal end enters a cavity in the carapace cuticle and reaches a terminal pore situated at the far end of the cavity. The cuticle above the cavity is modified. In both species the epicuticle is partly perforated by numerous small pores and the underlying exocuticle is much thinner and less electron dense than the regular exocuticle. Lattice organs very probably have a chemosensory function and are homologous with the sensory dorsal organ of other crustacean taxa. Accepted: 18 August 1998     

Cypris larvae of acrothoracican barnacles (Thecostraca: Cirripedia: Acrothoracica)

We used SEM to investigate the morphology of the cypris larvae from a range of species of the Cirripedia Acrothoracica, representing all three families and including the first detailed account of cyprids in the highly specialized Cryptophialidae. Special attention was given to the head shield (carapace), the lattice organs, the antennules, the thoracopods, the telson and the furcal rami. The cypris larvae of the Acrothoracica fall into two morphological groups; those of the Trypetesidae and Lithoglyptidae have a well-developed carapace (head shield) that can completely enclose the body and sports fronto-lateral pores, numerous short setae and lattice organs perforated by numerous small, rounded pores and a single, conspicuous terminal pore. The fourth antennular segment has the setae arranged in subterminal and terminal groups. There is a developed thorax with natatory thoracopods and a distinct abdomen and telson. In comparison, the cyprids of the Cryptophialidae exhibit apomorphies in the morphology of the carapace, the antennules and the thorax, mostly in the form of simplifications and reductions. They have a much smaller head shield, leaving parts of the body directly exposed. The shield is conspicuously ornamented by deep pits and hexagonally arranged ridges and bears a few, very long setae but lacks fronto-lateral pores. The lattice organs have numerous elongated pores, but no large, terminal pore. The fourth antennular segment has all the setae clustered in one terminal group. The thorax and thoracopods are rudimentary and not suitable for swimming. These reductions and simplifications in morphology correlate with cryptophialid cyprids being unable to swim. They can only disperse by antennular walking resulting in small, but highly gregarious populations of adults. The variations in antennular morphology and telson structure were traced for the genera of the families Lithoglyptidae and Trypetesidae. The traditional non-cladistic taxonomy in the suborders Pygophora (Cryptophialidae+Lithoglyptidae) and Apygophora (Trypetesidae) was based largely on symplesiomorphies in adult morphology and cannot be upheld. The Lithoglyptidae and Trypetesidae may form a monophylum, but evidence remains scarce. We expect that the use of larval (cyprid) characters will in the future play an important part in more detailed phylogenetic analyses of the Acrothoracica and also shed new light on their reproductive ecology.     

External morphology of the two cypridiform ascothoracid-larva instars of Dendrogaster: The evolutionary significance of the two-step metamorphosis and comparison of lattice organs between larvae and adult males (Crustacea, Thecostraca, Ascothoracida)

We describe the external morphology of the two cypridiform larval instars (first and second ascothoracid-larvae, or “a-cyprids”) of the ascothoracidan genus Dendrogaster. Ascothoracid-larvae of five species were studied with light and scanning electron microscopy, including both ascothoracid-larval instars in Dendrogaster orientalis Wagin. The first and second instars of the ascothoracid-larvae differ in almost all external features. The carapace of instar 1 has a smooth surface and lacks pores, setae, and lattice organs, while instar 2 has all these structures. The antennules of the first instar have only a rudimentary armament, the labrum does not encircle the maxillae, thoracopods 2-3 are not armed with a plumose coxal seta, and the abdomen is four-segmented (versus five-segmented in instar 2). Thus, the first ascothoracid-larva of Dendrogaster represents a transitional, generally brooded stage between the naupliar stages and the dispersive and fully functional second ascothoracid-larva that accomplishes settlement. The presence of two instars of ascothoracid-larvae (a-cyprids) in members of the order Dendrogastrida differs from the single cypridiform instar found in the Cirripedia (cyprid) and Facetotecta (y-cyprid), and we discuss the evolutionary significance of these ontogenies. We found lattice organs in both the second ascothoracid-larvae and in adult males of Dendrogaster. We could not observe both ascothoracid-larvae and males in any single species, but our data suggests that the lattice organs change significantly at the molt between these two instars. The lattice organs of second ascothoracid-larvae have no distinct keel and are situated in wide, shallow pits, whereas they have the ground pattern “crest-in-a-trough” morphology in adult males of two additional species examined for comparison. The positions of the terminal pores of lattice organs 1 and 2 also seem to change during maturation. These findings show that comparative data on lattice organ morphology for phylogenetic purposes must derive from strictly homologous instars, viz., the second ascothoracid-larva (a-cyprid) of the Ascothoracida, the y-cyprid of the Facetotecta, and the cyprid of Cirripedia. The ascothoracid-larvae of Dendrogaster and those of the family Ascothoracidae have four pairs of lattice organs, which suggests that this genus and family form a monophylum, to the exclusion of Ulophysema, which then brings into question the monophyly of the Dendrogastridae. Ulophysema is currently placed in the Dendrogastridae, but its second ascothoracid-larva has lattice organs of different and more plesiomorphic number and morphology. We briefly review lattice organ morphology across the Thecostraca. These organs are normally considered structures of the cypridiform larva and their presence in adult (males) Ascothoracida is unique in the Thecostraca. The continued morphological modification of these sensory structures in males compared to ascothoracid-larvae may suggest that they originated in adult thecostracans, but have come to be functional in the cypridiform larvae as well.     

Cypris morphology in the barnacles Ibla and Paralepas (Crustacea: Cirripedia Thoracica) implications for cirripede evolution

We used scanning electron microscopy (SEM) to describe cypris morphology in species of the barnacles Ibla and Paralepas, both of which are pivotal in understanding cirripede evolution. In Ibla, we also studied late naupliar stages with video and SEM. Special emphasis was put on the lattice organs, the antennules and the thorax and telson. In Paralepas we had settled specimens only and could therefore only investigate the carapace with the lattice organs. Cyprids of Ibla quadrivalvis and Paralepas dannevigi have five sets of lattice organs, grouped as two anterior and three posterior pairs. The organs are of the pore‐field type and the terminal pore is situated anteriorly in the first pair, just as in the Rhizocephala and the Thoracica. In Ibla the armament of antennular sensilla resembles that found in the Thoracica but differs from the Rhizocephala. The absence of setules on the A and B setae sited terminally on the fourth antennular segment is a similarity with the Acrothoracica. The attachment disc is angled rather than facing distally and is encircled by a low cuticular velum. The thoracopods have two‐segmented endopods and exopods as in the Thoracica, but the number, shape, and position of thoracopodal setae differ somewhat from other species of that superorder. Both Ibla and Paralepas cyprids have a deeply cleaved telson, but no independent abdominal part. In cypris morphology, Ibla and Paralepas show several synapomorphies with the clade comprising Rhizocephala and Thoracica and there are no specific apomorphies with either the Acrothoracica, the Rhizocephala or any particular subgroup within the Thoracica. This is in agreement with recent molecular evidence that Ibla (Ibliformes) is the sister taxon to all other Thoracica and the ibliforms therefore become the outgroup of choice for studying character evolution within the superorder. Paralepas, and other pedunculated barnacles without shell plates, are apparently not primitive but are secondarily evolved and nested within the Thoracica. J. Morphol., 2009. © 2008 Wiley‐Liss, Inc.     

Deep phylogeny and character evolution in thecostraca (crustacea: maxillopoda)

The thecostracans include the Facetotecta, Ascothoracida, and Cirripedia and show great diversity in both morphology and biology. This makes them ideal models for studying evolutionary adaptations of the larval and adult body-plan, lifestyle, and reproduction. Surprisingly, despite all the work published since Darwin's seminal monographs, few studies have tested evolutionary hypotheses about Thecostraca within a phylogenetic context. In this review, we combine a Bayesian phylogenetic method and multilocus sequence data to reconstruct the evolutionary history of 12 key thecostracan phenotypic traits associated with their lifecycle, larval biology, reproduction, and adult morphology. Our analyses show that thecostracan biological diversity resulted both from unique innovations and from events of convergence. This provides an opportunity to reevaluate previous classifications of the Thecostraca and the theories relating to the origin and diversification of this taxon.     

Remarkable convergent evolution in specialized parasitic Thecostraca (Crustacea)

Background  

The Thecostraca are arguably the most morphologically and biologically variable group within the Crustacea, including both suspension feeders (Cirripedia: Thoracica and Acrothoracica) and parasitic forms (Cirripedia: Rhizocephala, Ascothoracida and Facetotecta). Similarities between the metamorphosis found in the Facetotecta and Rhizocephala suggests a common evolutionary origin, but until now no comprehensive study has looked at the basic evolution of these thecostracan groups.     

The Phylogenetic Position of the Rhizocephala: Are They Truly Barnacles?

Incorporation of the Rhizocephala in the Cirripedia, reflecting the traditional view that these parasites evolved from a setose feeding barnacle, has recently been challenged in favour of rhizocephalans being the sister group to all other Thecostraca or a scenario where they evolved from a free-living, ‘precirripede’ ancestor. Adult morphology is useless in discussing the monophyly of the Cirripedia, since rhizocephalan adults are too reduced to furnish any phylogenetic evidence. But numerous, detailed similarities in nauplii and cyprids of the Thoracica, Acrothoracica and Rhizocephala as well as the ultrastructure of their sperm are synapomorphic relative to other Thecostraca and indicate that these three orders form a monophylum. There is evidence that the stylet in the rhizocephalan kentrogon is homologous to an element in the ancestral mouth field. If so, the Rhizocephala probably evolved before setose feeding was adopted, and constitute the sister group to the Acrothoracica and Thoracica. This conclusion is based on frail evidence so the term Cirripedia should be retained to comprise the Rhizocephala, Thoracica, and Acrothoracica. These three orders all possess remarkably similar cyprids, adapted to accomplish irreversible settlement by cement secretion and initiate metamorphosis, so their last common ancestor was most probably a permanently sessile organism.     

When similar beginnings lead to different ends: Constraints and diversity in cirripede larval development

Summary

Cirripedes are fascinating models for studying both functional constraints and diversity in larval development. Adult cirripedes display an amazing variation in morphology from sessile suspension feeders that still retain many crustacean characters to parasites that have lost virtually all arthropod traits. In contrast, cirripede larval development follows a common scheme with pelagic larvae comprising a series of nauplii followed by a cyprid. Variations are mostly concerned with whether or not the nauplii are feeding and the degree of abbreviation of development, culminating in species where the larvae hatch as cyprids. The cypris larvae are very similar among the ingroups of the Cirripedia, but interesting variations occur in structures used for substrate location and attachment. The cyprid is specialized to both swim through the water and actively explore the substratum by walking on the antennules and using an array of sensory organs in search for a suitable site to attach. This unique morphology and behavior of the cyprid have enabled the Cirripedia to colonize widely different habitats ranging from hard rock to soft animal tissue. Yet, the cyprid can metamorphose into juveniles as different as a setose feeding barnacle and the vermiform stages of the parasitic forms. This emphasizes the importance of the cyprid as one of the key features for the evolutionary success of the Cirripedia.     

Developmental Patterns and Hypotheses of Homology in the Antennules of Thecostracan Nauplius Larvae (Crustacea)

Abstract Antennular development is summarized for non–brooded nauplius larvae of the Ascothoracida (families Lauridae and Petrarcidae) and for selected nauplii of the Facetotecta. In the Cirripedia Thoracica, nauplii of the Lepadomorpha and Scalpellomorpha/Sessilia are shown to differ in their patterns of antennular setal addition and division into articles, and one or two possible apomorphies for each group are identified. Several variations of the scalpellomorphan/sessilian pattern are outlined. These data on the Ascothoracida, Facetotecta, and Cirripedia provide the basis for modifying an earlier proposal of structural homologies among the naupliar antennules of thecostracan maxillopodans. ‘Virtual’ articles (ones that are never completely free) are invoked to account for setae that ‘jump’ across article boundaries at molts and for supposedly homologous setae found on adjacent articles in different taxa. The identities of the preaxial setae in the Cirripedia are ambiguous and force consideration of two models for that group and for the thecostracan antennular Bauplan; the latter may have included up to 11 or 12 articles, more than the previously supposed 8 or 9. The scoring of characters used in an earlier cladistic study of thecostracan and other maxillopodan taxa is not seriously affected. A comparison with the Cambrian maxillopodan Bredocarissuggests that division of the antennule into discrete articles in extant thecostracans is an apomorphic state and that yet one more apical article once existed in the maxillopodan antennule.     

Gregarious settlement in cypris larvae:the effects of cyprid age and assay duration

Gregarious behaviour of marine larvae is perhaps most clearly associated with finding a suitable habitat in a changeable or restricted environment, or with finding other conspecifics with which to mate. Prior work has shown that in settlement assays using cypris larvae of the barnacle Balanus amphitrite, gregarious interactions significantly affected the interpretation of experiments testing the activity of organic settlement promoters and inhibitors. Other studies have also shown effects of cyprid age and pheromone concentration on settlement behaviour. However, the effects of interactions between gregariousness and these two factors are not known. The aim of this study was to test the hypotheses that i) as cyprids age the effects of gregariousness become less apparent, and ii) as the duration of the experiment increases gregarious effects become more apparent, using cypris larvae of B. amphitrite and Balanus improvisus. Three age classes of cyprids were used at six densities in a fully factorial design. For B. improvisus cyprids significant gregarious effects occurred between 3 or more larvae, and although larval age and experiment duration had significant main effects, there were no interactions between these important factors and gregariousness. For B. amphitrite cyprids significant gregarious effects also occurred with 3 larvae per well, though this effect was strongly dependent upon experiment duration. B. amphitrite cyprid sensitivity to conspecific cues does not change with age, although increasing experiment duration and age interact to increase settlement. Differences between species may be due to different thresholds to conspecific larval cues, or B. improvisus cyprids release much more larval temporary adhesive during exploration.     

The setae of parasitic Liphyra brassolis butterfly larvae form a flexible armour for resisting attack by their ant hosts (Lycaenidae: Lepidoptera)

Caterpillars of the lycaenid butterfly, Liphyra brassolis, live inside the nests of arboreal weaver ants, Oecophylla smaragdina, and eat their brood. Observations of mature larvae suggest that they are impervious to relentless ant molestation, yet they lack sclerotized cuticular plates. We document a novel form of integumental defence that imparts protection from ant attack whilst maintaining the flexibility necessary to walk with a hydraulic skeleton. Analysis of the trunk integument and cuticular structures of early and late instars of L. brassolis using light microscopy, scanning electron microscopy, and histology revealed three new setae types (disc, clavate, and lanceolate), as well as three new cuticular structures (pored sockets, cuticular pores, and cuticular domes). The unique cuticle is covered with lanceolate setae, which act as endocuticular struts, and overlapping scale‐like sockets, which form a hard, flexible integument. The imperfect armour of the early‐instar larvae suggests that abundant, putatively secretory pores are likely to be homologous to pore cupola organs (PCOs) found in other lycaenid larvae and thus may exude semiochemicals to allay ant aggression. The importance of these pores presumably wanes as structural (setal) cuticular defenses are reinforced in later instars, when adult ants have been observed attacking caterpillars to no avail. The caterpillar's antennae are unusual and seem to be involved in manipulating ant larvae into the caterpillar's mouth. Behavioural observations indicate that the dexterity of these structures is associated with eating ants (myrmecophagy).     

Identification of methyl farnesoate in the cypris larva of the barnacle, Balanus amphitrite, and its role as a juvenile hormone

Previous investigations have shown that insect juvenile hormone (JH) and its analogues induce precocious metamorphosis of barnacle cypris larvae. In the present study, methyl farnesoate (MF; structurally identical to JH III, except for the absence of an epoxide group) has been shown to have a concentration-dependent effect on the development of cyprids of the barnacle Balanus amphitrite. Analysis of cypris extracts by gas chromatography-mass spectrometry with selected ion monitoring (GC-MS-SIM) confirmed the presence of endogenous MF. These data provide evidence that MF functions as a juvenilizing hormone in barnacle cyprids, an effect that hitherto has not been noted.     

On the morphology of antennular sensory and attachment organs in cypris larvae of the deep‐sea vent/seep barnacles,Ashinkailepas and Neoverruca

Barnacle cypris larvae show high morphological variation in the organs used in search of and attaching to a substratum. This variation may represent adaptation to the habitat of the species. Here, we studied SEM level morphologies of cypris antennular sensory and attachment organs in a deep‐sea vent endemic species (Neoverruca sp.) and a vent/seep inhabiting species (Ashinkailepas seepiophila). We compare them with three species from other environments. The antennular morphologies of Neoverruca sp. and A. seepiophila were similar, which is consistent with recent molecular studies showing a close relationship of the two species. The setation pattern of the antennules was very conservative among species from various environments. In contrast, striking differences were observed in the structure of the attachment organ (the third antennular segment). Neoverruca sp. and A. seepiophila had no velum or a skirt surrounding the attachment disc on the third segment, while other cirripede cyprids almost always have either of these structures. In addition, both cyprids of A. seepiophila and Neoverruca sp. had the attachment disc angled toward the substratum, whereas it faces distally in cyprids from hard bottom inhabiting barnacles. We suggest that both velum/skirt and the angle of the attachment disc play an important role, when the antennules are contacting the substratum during surface exploration. Differences in attachment organ structures may be highly adaptive, enabling cirripede species to enter new habitats during evolution. J. Morphol. 277:594–602, 2016. © 2016 Wiley Periodicals, Inc.     

Size and settling behaviour in male and female cypris larvae of the parasitic barnacle Sacculina carcini Thompson (Crustacea: Cirripedia: Rhizocephala)

Twenty-one broods from different externae of Sacculina carcini Thompson were cultured to the cypris stage. The size of the cyprids was measured and the larvae subjected to settling upon unparasitized crabs (Garcinus maenas (L.)) and small juvenile externae. The cyprids occur in two sizes that may appear singly or together in the same brood. Small cyprids are of the female sex that settle upon crabs and are infective, while large cyprids are of the male sex and only settle upon juvenile externae. These results are in agreement with other well-studied rhizocephalans.     

Combined effects of temperature and salinity on larval development and attachment of the subtidal barnacle Balanus trigonus Darwin

The combined effects of temperature and salinity on larval development and attachment of Balanus trigonus Darwin (Cirripedia, Balanidae) was examined under controlled laboratory conditions. Whilst larval survivorship was not affected (>70%), the duration of larval development was significantly affected by temperature and salinity. The effect of temperature was comparatively stronger than that of salinity. The majority of nauplius II larvae metamorphosed into cypris stage after 4-5 and 10-11 days at 28 °C (22-34‰) and 18 °C (22-34‰), respectively. Temperature, salinity and the duration of assay had a significant effect on cypris attachment with significant interaction among these main effects. Maximum (>80% in 6 days) and minimum percent attachment (0% in 6 days) on polystyrene surfaces were observed at 24 °C (34‰) and 18 °C (22‰), respectively. At high temperature (28 °C) and low salinity (22-26‰), larvae rapidly (4 days) developed into cyprids, but less than 33% attached. These results suggest that low larval attachment rates may lead to the low recruitment of B. trigonus in Hong Kong waters during summer when the water temperature is high (about 28 °C) and salinity is low (<26‰).     

Origin of the Ostracoda and their maxillopodan and hexapodan affinities

There are Cambrian fossils attributed to the Ostracoda but the extant subclasses Podocopa and Myodocopa do not appear until the Ordovician. At this time the morphologically similar, free-living ancestors of the now sedentary Thecostraca (Ascothoracida, Acrothoracica and Cirripedia) may have still been extant, and from an ecological point of view it seems likely that, by and large, ostracods replaced them. However, living ostracods have an abbreviated, direct development, and some key aspects of their morphology, such as the nature of the maxillary segment and abdomen, are conjectural. Thus the affinities between these and related taxa remain uncertain; e.g., while some contemporary carcinologists place Ostracoda as a taxon coordinate with the Branchiopoda, Remipedia, Cephalocarida, Maxillopoda, Malacostraca, others tentatively or unequivocally ally them with the Maxillopoda (generally Mystacocarida, Copepoda, Tantulocarida and Thecostraca, and sometimes Branchiura and Pentastomida). Others, largely involved with fossils, have stretched the definition of the Maxillopoda even further, to the point where it seems even less likely a monophyletic taxon. Until recently cladistic analyses utilizing genetic (largely 18S rDNA) as well traditional morphological characteristics have given confusing results regarding the affinities between these taxa, and an important one suggested the Ostracoda might even be diphyletic. Furthermore, a very recent genetic study utilizing protein encoding genes places a podocopine ostracod among the most primitive of the extant crustaceans (Branchiopoda, Cephalocarida Remipedia and Mystacocarida), and then generally at the base of a lineage leading to the Malacostraca, a lineage giving rise to copepods and cirripeds along the way. This indicates these so-called maxillopodan taxa evolved independently from a malacostracan-like ancestor, and if so they are convergent. And finally, from genetic studies it is not only becoming well documented the Crustacea rather than Myriapoda gave rise to the Hexapoda, but it appears the Hexapoda stem from among the lower rather than the higher crustaceans, possibly even from the Ostracoda. Whether there were terrestrial ostracods at the time hexapods appeared in the Lower Ordovician is unknown, but the modest diversity of terrestrial ostracods today are podocopines which also first appeared in the Lower Ordovician. Thus, if current interpretations of living ostracodan and fossil hexapodan body plans are largely correct, it can be hypothesized the Ostracoda are close to the ancestor of the Hexapoda.     

The relation between cypris ultrastructure and metamorphosis in male and female Sacculina carcini (Crustacea,Cirripedia)

Summary To elucidate current controversies on sex in rhizocephalan barnacles, broods of Sacculina carcini, infesting the shore crab Carcinus maenas, were raised to cyprids in the laboratory and followed through settlement and metamorphosis. Free-swimming cyprids were studied by transmission electron microscopy and occur in male and female morphological types, which differ in the structure of carapace cuticle, antennular cuticle, antennular glands, and the cells suspected of being the stem cells during metamorphosis. These dissimilarities are in addition to the already known differences in cypris size, in number of antennular sense organs, and in substrata settled on by morphological males and females. Metamorphosing males (trichogons) and females (kentrogons) are illustrated in interference phase-contrast micrographs. The morphological differences between male and female cyprids are directly related to their dissimilar metamorphosis. Hence, cyprids of male morphology are anatomically incapable of metamorphosing into kentrogons, while cyprids of female morphology cannot metamorphose into trichogons. The determination of sex in rhizocephalan barnacles is discussed.The results refute the hypothesis that sex in Sacculina carcini is determined environmentally, e.g., by the substratum encountered by the cyprids at settlement. It is concluded that sex is determined already in the free-swimming larvae and, most probably, already in the ovary. This agrees with the mode of sex determination in other species of the Rhizocephala Kentrogonida.