Not Yet Proven Crustacea: the Thylacocephala

The Thylacocephala is a recently recognized extinct group of uncertain, but probably crustacean, affinity, comprising fossils formerly assigned to many different crustacean groups. Key characters are three anterior pairs of subchelate limbs; at least 8 pairs of radiating gills, situated between the trunk and carapace, the most posterior gill lying subhorizontally; and the large posteriorly tapering battery of 8(–?20) elongated limb protopods with tenuous distal filaments. Thylacocephala bear huge anterior structures, variously identified as compound eyes or as a ‘cephalic sac’. Microsclerites borne on this structure may be external, perhaps setae, but need not imply affinity with cirripedes as has been argued. Internal microsclerites have been compared with crystalline cones of a compound eye. More detailed study of well-preserved material is required to resolve the nature of these microsclerites, to elucidate details of appendages, to locate antennae, and to ascertain limb homologies. Until that work is done, affinities of the Thylacocephala must remain uncertain as must the group's relevance to evolution of Crustacea. Such recent recognition of a distinctive functional model system (Dahl, E. 1976. Zoologica Scripta 5: 163–166) which persisted for at least 340 million years, shows the fossil record still has much to contribute to understanding the pattern of arthropod evolution.     

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     

Structure and cuticle formation of the reticulated carapace of the ostracode Bicornucythere bisanensis

Electron microscopic examination revealed fine structures of the adult carapace of Bicornucythere bisanensis , which consists mainly of epicutiele, procuticle, membranous layer, epidermal cells and subdermal cells. Piled membrane structure is recognized as an organic framework of the procuticle. Newly formed cuticle examined with specimens at various stages of the molt cycle clarified the process of cuticle formation. Analysis of the complicated process of cuticular ridge formation permits morphogenetic discussion of surface features of the ostracode carapace of many species. Cuticle formation in close relation to cell boundaries clearly indicates that each polygon of the carapace reticulation is formed by one epidermal cell. The cell-polygon correspondence suggests that ostracode reticulation is important for elucidation of the phylogenic and ontogenic development of multicellular organisms at cell level.
□ Ostracoda, carapace structure, biomineralization, cuticle formation.     

Carapace formation of the podocopid ostracode Semicytherura species (Crustacea: Ostracoda)

Details of ostracode carapace structures were examined by SEM and TEM. The podocopine ostracode Semicytherura kazahana has major ridges on the carapace surface and develops its prismatic layer inside the adult carapace. Electron microscopy at the final molt reveals that the major ridges arise from the highly dense formation of pits within the underlying swollen epidermis, and that disappearance of the epidermis in the presumptive area of the prismatic layer occurs after the calcification of the outer lamella cuticle, and just before synthesis of the membranous layer. These facts suggest that the formation of the carapace in Semicytherura takes place via a more complex process than that of the other podocopid ostracodes.     

Thylacocephala (Arthropoda: Crustacea?) from the Cretaceous of Lebanon and implications for thylacocephalan systematics

An intensive study of a collection of arthropods from the Cretaceous of Lebanon, formerly referred to as stomatopod larvae, reveals that these forms belong to the problematical arthropod class Thylacocephala. The species Protozoea hilgendorfi , P. damesi , and Pseuderichthus cretaceus display defining thylacocephalan characters such as a carapace enclosing the entire body bearing a large anterior optic notch; three pairs of large, raptorial appendages; and a posterior battery of small swimming limbs associated with muscle segments. Unique characters of at least the genus Protozoea are the numerous 'pits' covering the entire carapace, elongated anterior rostral and posterior spines, and a dorsal hinge indicative of a truly bivalved carapace. We note several anatomical curiosities that provide some insight into the anatomy, ecology and phylogenetic relationships of these and other thylacocephalan species. Structures on the carapace surface of thylacocephalans may represent sensory organs comparable to the receptors seen on thecostracan crustaceans. The affinities of the three species described are still not entirely clear, but a phylogenetic analysis of the entire group may resolve this.     

Cuticle morphogenesis in crustacean embryonic and postembryonic stages

The crustacean cuticle is a chitin-based extracellular matrix, produced in general by epidermal cells and ectodermally derived epithelial cells of the digestive tract. Cuticle morphogenesis is an integrative part of embryonic and postembryonic development and it was studied in several groups of crustaceans, but mainly with a focus on one selected aspect of morphogenesis. Early studies were focused mainly on in vivo or histological observations of embryonic or larval molt cycles and more recently, some ultrastructural studies of the cuticle differentiation during development were performed. The aim of this paper is to review data on exoskeletal and gut cuticle formation during embryonic and postembryonic development in crustaceans, obtained in different developmental stages of different species and to bring together and discuss different aspects of cuticle morphogenesis, namely data on the morphology, ultrastructure, composition, connections to muscles and molt cycles in relation to cuticle differentiation. Based on the comparative evaluation of microscopic analyses of cuticle in crustacean embryonic and postembryonic stages, common principles of cuticle morphogenesis during development are discussed. Additional studies are suggested to further clarify this topic and to connect the new knowledge to related fields.     

How do mandibles sense? – The sensory apparatus of larval mandibles in Palaemon elegans Rathke, 1837 (Decapoda,Palaemonidae)

The mandibles of decapod zoea-I larvae are robustly built masticating mouthparts equipped with several processes and spines. Superficial examination of these sturdy, inflexible structures can suggest that they are lacking sensory receptors. However, detailed TEM analysis of their ultrastructure revealed up to 11 sensillar cell clusters on the gnathal edges of the mandibles of the zoea-I in Palaemon elegans Rathke, 1837. Based on ultrastructural criteria we distinguish 7 types of sensilla: mechanoreceptors, chemoreceptors and mechano- and chemoreceptors. One sensory unit located at the base of the ‘lacinia mobilis’ exhibits the typical features of a crustacean mechanosensitive sensillum with an external seta and corresponding ultrastructure. Another unit shows features indicating bimodal contact chemosensitivity. A third one is similar to known olfactory chemoreceptors.Using the concept of modality-specific structures we analyse the structure and functional morphology of each sensillum, and give a comprehensive overview of the sensory abilities of zoea mandibles. We take a closer look at the ultrastructure of the ‘lacinia mobilis’, providing further features to trace its evolutionary history in Decapoda, and thus contributing to a better understanding of malacostracan phylogeny.     

Ultrastructural investigation of the nuchal organs ofPygospio elegans (Polychaeta). I. Larval nuchal organs

The structural differentiation of the nuchal organs during the post-embryonic development ofPygospio elegans is described. The sensory organs are composed of two cell types: ciliated cells and bipolar primary sensory cells, constituting the nuchal ganglion, which is associated with both the sensory epithelium and the brain. Since the sensory neurons are largely integrated into posterolateral parts of the cerebral ganglion, the nuchal organs are primary presegmental structures. The microvilli of the ciliated cells form a cover over the cuticle with a presumed protective function. An extracellular space extends between cuticle and sensory epithelium. The distal dendrites of the sensory cells terminate in sensory bulbs, bearing one modified sensory cilium each that projects into the olfactory chamber, embedded within the secretion of the ciliated cells. During development, the nuchal organs increase in size. This is accompanied by a shift in position, an expansion of the sensory area, and secretory activity of the ciliated cells. The nuchal ganglion differentiates into three nuchal centres forming three distinct sensory areas around the ciliated region. Each nuchal complex reveals two short nuchal nerves comprising the sensory axons, which enter the posterior circumesophageal connective. The sensory cells lying in the brain exhibit neurosecretory activity; the sensory cilia enlarge their surface area by dilating and branching. Nuchal organs accomplish the basic structural adaptions of chemoreceptors and show structural analogies to arthropod olfactory sensilla; thus, there is every reason to suppose chemoreceptor function.     

Butterflies of the Cambrian benthos? Shield position in bradoriid arthropods

Mode of preservation and method of recovery strongly influences our understanding of the life habits of extinct organisms. Bradoriid arthropods were abundant, and diverse members of early Cambrian ecosystems and most life reconstructions display these animals with the two shields of the carapace open in a ‘butterfly’ configuration. This favoured reconstruction is largely based on the abundance of ‘crack‐out’ specimens preserved in this position (e.g. Kunmingella from the early Cambrian of China). In contrast, large collections of acid processed bradoriids from the Arrowie Basin of South Australia (Cambrian Stage 3) are preserved with a narrow gape at the ventral margin or completely closed with the carapace folded along the dorsal midline. The relative abundance of conjoined, closed (or partially closed) specimens from the lower Cambrian Hawker Group succession suggests that at least some bradoriid taxa were capable of withdrawing appendages and tightly closing the shields, challenging the common view that the majority of bradoriids usually held their carapaces open in a ‘butterfly’ configuration during life. New data show that layers of the bradoriid carapace are continuous through the dorsal fold with no evidence for complex articulating structures as in ostracod hinges. The relatively pliable, sclerotized or lightly mineralized calcium phosphate composition of the carapace and the simple, flexible dorsal fold facilitated opening and closing of the shields. Despite not being closely related, ostracods share close biomechanical and ecological similarities with bradoriids. The evolution of more complex articulating hinge structures – together with well‐developed musculature – in ostracods during the Early Ordovician, may have provided more efficient means for shield articulation and movement, thus promoting the ecological success of ostracods throughout the Phanerozoic.     

Molecular composition of the wall of insect olfactory sensilla-the chitin question

Identification of chitin in sensory hairs of olfactory sensilla of silkmoths was performed using two independent methods. Firstly, ultrathin sections were labelled with gold-conjugated wheat germ agglutinin and showed positive labelling in the cuticule of sensilla as well as in the antennal cuticle. Secondly, isolated sensory hairs and body scales were subjected to analytical pyrolysis in combination with gas chromatography and mass spectrometry. Chromatograms of both sensory hairs and scales, included several pyrolysis products, which unequivocally demonstrate the contribution of chitinous moieties to the chemical composition of both types of cuticle. This study supports the notion that even the very thin cuticle of olfactory sensilla is composed of both an epi- and a true exocuticle. The carbohydrate components of the latter cuticle most probably are responsible for the extremely high resilience and breaking limit of these delicate structures.     

Revising the definition of the crustacean seta and setal classification systems based on examinations of the mouthpart setae of seven species of decapods

The crustacean cuticle has numerous projections and some of these projections, the setae, have important mechanical as well as sensory functions. The setae display a wide diversity in their external morphology, which has led to great problems separating setae from other projections in the cuticle and problems in making a consistent classification system. Here, the cuticular projections on the mouthparts of seven species of decapods are examined by scanning and transmission electron microscopy. A new definition is given: a seta is an elongate projection with a more or less circular base and a continuous lumen; the lumen has a semicircular arrangement of sheath cells basally. From the details of the external morphology the mouthpart setae are divided into seven types: pappose, plumose, serrulate, serrate, papposerrate, simple and cuspidate setae, which are suggested to reflect mechanical functions and not evolutionary history. This classification system is compared with earlier systems.  © 2004 The Linnean Society of London, Zoological Journal of the Linnean Society , 2004, 142 , 233–252.     

The crustacean cuticle does not record chronological age: New evidence from the gastric mill ossicles

A proposed method to determine chronological age of crustaceans uses putative annual bands in the gastric mill ossicles of the foregut. The interpretation of cuticle bands as growth rings is based on the idea that ossicles are retained through the moult and could accumulate a continuous record of age. However, recent studies presented conflicting findings on the dynamics of gastric mill ossicles during ecdysis. We herein study cuticle bands in ossicles in four species of commercially important decapod crustaceans (Homarus gammarus, Nephrops norvegicus, Cancer pagurus and Necora puber) in different phases of the moult cycle using dissections, light microscopy, micro-computed tomography and cryo-scanning electron microscopy. Our results demonstrate that the gastric mill is moulted and ossicles are not retained but replaced during ecdysis. It is therefore not plausible to conclude that ossicles register a lifetime growth record as annual bands and thereby provide age information. Other mechanisms for the formation of cuticle bands and their correlation to size-based age estimates need to be considered and the effect of moulting on other cuticle structures where ‘annual growth bands’ have been reported should be investigated urgently. Based on our results, there is no evidence for a causative link between cuticle bands and chronological age, meaning it is unreliable for determining crustacean age.     

Exoskeletal cuticle differentiation during intramarsupial development of Porcellio scaber (Crustacea: Isopoda)

Exoskeletal crustacean cuticle is a calcified apical extracellular matrix of epidermal cells, illustrating the chitin-based organic scaffold for biomineralization. Studies of cuticle formation during molting reveal significant dynamics and complexity of the assembly processes, while cuticle formation during embryogenesis is poorly investigated. This study reveals in the terrestrial isopod Porcellio scaber, the ultrastructural organization of the differentiating precuticular matrices and exoskeletal cuticles during embryonic and larval intramarsupial development. The composition of the epidermal matrices was obtained by WGA lectin labelling and EDXS analysis. At least two precuticular matrices, consisting of loosely arranged material with overlying electron dense lamina, are secreted by the epidermis in the mid-stage embryo. The prehatching embryo is the earliest developmental stage with a cuticular matrix consisting of an epicuticle and a procuticle, displaying WGA binding and forming cuticular scales. In newly hatched marsupial larva manca, a new cuticle is formed and calcium sequestration in the cuticle is evident. Progression of larval development leads to the cuticle thickening, structural differentiation of cuticular layers and prominent cuticle calcification. Morphological characteristics of exoskeleton renewal in marsupial manca are described. Elaborated cuticle in marsupial larvae indicates the importance of the exoskeleton in protection and support of the larval body in the marsupium and during the release of larvae in the external environment.     

Transmission and scanning electron microscopic studies of crustacean shell disease in fish lice of the genus Argulus (Crustacea: Branchiura)

Juvenile and adult specimens of the branchiuran crustacean parasite, Argulus foliaceus , have been found to exhibit the brown areas of damaged cuticle typical of crustacean shell disease. Transmission and scanning electron microscopy of such lesions at a variety of sites on the body surface have demonstrated that they harbour dense populations of bacteria and occasionally fungi, in zones of superficial cuticular erosion. Complete penetration of the exoskeleton was inferred in instances when circular lesions were associated with bacteraemia of underlying soft tissues.     

Sensory structures of Archaeostraca (Phyllocaridida, Crustacea)

SUMMARY The carapace of some Archaeostraca exhibits two kinds of structures that are best interpreted as corresponding to sensory organs. Dorsal ones are located either along the dorsal margin of the valves of the carapace, or on the median dorsal plate. Anterolateral ones are located in the anterior region of both valves. Both structures are respectively homologous of the anterior and posterior sensory organs known from extant Leptostraca and other Crustacea. These structures are described in adult Archaeostraca from the Ordovician to the Carboniferous. It is expected that they could have baroreceptor, mechanoreceptor, and/or chemoreceptor functions in some cases, and a function in osmotic regulation in others.     

Early pattern of calcification in the dorsal carapace of the blue crab, Callinectes sapidus

The pattern of calcium carbonate deposition was observed in the dorsal carapace of premolt (D2-D3) and early postmolt (0-48 h) blue crabs, Callinectes sapidus, using scanning (SEM) and transmission (TEM) electron microscopy. Samples of dorsal carapace for SEM were quick-frozen in liquid nitrogen, subsequently lyophilized, and viewed using secondary and backscattered electrons as well as X-ray maps of calcium. Pieces of lyophilized cuticle were also embedded in epoxy resin and subsequently sectioned and viewed with TEM and SEM. Fresh pieces of dorsal carapace for TEM were also fixed in 2.5% glutaraldehyde in phosphate buffer followed by postfixation in 1% OsO4 in cacodylate buffer. Calcium concentrations were determined using atomic absorption spectrophotometry and quantitative X-ray microanalysis. Calcium accumulation began in the cuticle at 3 h postmolt at the epicuticle/exocuticle boundary and at the distal and proximal margins of the interprismatic septa (IPS). The bidirectional calcification of the IPS continued until the two fronts met at 5-8 h postmolt. The roughly hexagonal walls of the IPS formed a honeycomb-like structure that resulted in a rigid cuticle. The walls of the canal containing sensory neurons also calcified at 3 h, thereby imparting rigidity to the structure and additional strength to the cuticle. Examination of thin sections of lyophilized cuticle and fixed cuticle revealed that the first mineral deposited is more soluble than calcite and is probably amorphous calcium carbonate. The amorphous calcium carbonate is transformed to calcite along a front that follows the original deposition and is probably controlled by a specialized matrix within the IPS. Since amorphous calcium carbonate is isotropic, it would also make the mineral in the exocuticle stronger by an equal distribution of mechanical stress.     

Electron microscopic and preparative methods for the analysis of isopod cuticle

The crustacean cuticle consists of a complex organic matrix and a mineral phase. The physical and chemical properties of the cuticle are corellated to the specific functions of cuticular elements, leading to a large variety in its structure and composition. Investigation of the structure-function relationship in crustacean cuticle requires sophisticated methodological tools for the analysis of different aspects of the cuticular architecture. In the present paper we report improved preparation methods that, in combination with various electron microscopic techniques, have led to new insights of cuticle structure and composition in the tergite cuticle of Porcellio scaber. We used thin sections of non-decalcified tergites and decalcified resin embedded material for transmission electron microscopy and scanning transmission electron microscopy. Etched sagittal planes of bulk tergite samples were analysed with field emission scanning electron microscopy. We have found a distinct distal region within the exocuticle that differs from the subjacent proximal exocuticle in the arrangement of fibres. Within this distal exocuticle chitin-protein fibrils assemble to fibres with diameters between 15 and 50 nm that are embedded in a mineral matrix. In the proximal exocuticle and the endocuticle fibrils do not assemble to fibres and are surrounded by mineral individually. Furthermore, we show that the pore canals are filled with mineral, and demonstrate that mild etching of polished sagittal cuticle surfaces reveals regions containing mineral of diverse solubility.     

The structure and development of a surface pattern on the cuticle of the green vegetable bug Nezara viridula

Adult Nezara possess an area of cuticle on the metathoracic sternite with a surface pattern of minute mushroom-like projections. The surface pattern is formed by two types of cell, one forming projections and the other depressions. The surface pattern is largely determined before epicuticle deposition by deformation of the epidermal surface. This is brought about by interactions between the moulting fluid secretions and the cells. Maintenance of shape of the cells may be associated with the presence of oriented cytoplasmic microtubules. The mushroom area may function as an external reservoir causing retarded evaporation of defensive secretions.     

Ultrastructure of the water-movement-sensitive sensilla in the medicinal leech

Behavioral and physiological experiments have shown that medicinal leeches are able to detect low amplitude surface waves, and further, that the transduction of this stimulus modality occurs primarily, if not exclusively, at the annular sensilla (Young, Dedwylder, and Friesen, 1981; Friesen, 1981). Here we examine the morphology of these specialized sensory structures using light, scanning electron, and transmission electron microscopes. We found that three types of ciliated sensory cells occur at the sensilla: (1) a uniciliate cell, with an axial cilium that projects at least 12 μm beyond the cuticle; (2) a multiciliate cell with from two to four grouped cilia that extend 1–3 μm beyond the cuticle; and (3) a second multiciliate cell, whose cilia project parallel to the body surface but remain within the cuticle. The cilia of all three cell types arise from the cuplike depressions which form the apices of slender, elongated cells (approximately 2 μm diameter × 50 μm length). A complexly interconnected ring of microvilli surrounds the cilium of the uniciliate cells. The morphology of the uniciliate cells closely resembles the structure of vibration-sensitive sensory neurons found in other species. We propose, based on previous results and our new findings, that the uniciliate receptor cells are the sensillar movement receptors which mediate leech sensitivity to water movements.