Occurrence of immunoreactive enkephalins in a neurohemal organ and other nervous structures in the eyestalk of the shore crab,Carcinus maenas L. (Crustacea,Decapoda)

Summary Light-microscopical observations with immunofluorescence and peroxidase staining procedures revealed leu-enkephalin-like immunoreactivity in axon profiles of the sinus gland (SG) and in single small neurons in the optic ganglia of the eyestalk of Carcinus maenas. Electron microscopy of the SG showed reactivity to be associated with neurosecretory granules 82±23 nm in diameter. High performance liquid chromatography of SG-extracts revealed radioimmunoreactive substances with the retention times of synthetic met- and leu-enkephalin and met-enkephalin-Arg⁶-Phe⁷, respectively.     

Ultrastructure of the Y-organ of Astacus astacus (L.) (Crustacea) in relation to the moult cycle

Summary The electron microscopical investigation of Y-organs of Astacus astacus revealed that during intermoult (stage C) the cytoplasm is poorly developed and that it increases at premoult (stage D). It then shows the typical signs of steroid production, namely agranular endoplasmic reticulum and mitochondria of the tubular type. Furthermore, a larger type of mitochondria with a regular pattern of internal structure is described.Supported by Sächsische Akademie der Wissenschaften zu LeipzigWe are grateful for technical assistance to Mrs. B. Cosack und Mrs. A. Schmidt     

Ultrastructural shape and three-dimensional organization of the intracuticular canal systems in the mineralized cuticle of the green crab Carcinus maenas

Two main self-contained canal systems are present in the crab mineralized cuticle. The first, or fibre canal system, is constituted by simple, unbranched vertical canals containing axially running fibres in close association with myoepidermal junctions. The second, or pore canal system, is composed of procuticular pore canals and epicuticular channels that prolong the procuticular canals. In opposition to widespread opinion, pore canals make up a three-dimensional branched system extending from the apical plasma membrane of the epidermis up to the epicuticle. Branching occurs by projections of lateral horizontal from the vertical canals at the lower level of the pigmented layer and by innumerable ramifications of epicuticular canals. In agreement with Neville's model for insects, vertical procuticular pore canals of crustacean mineralized cuticle, and also fibre canals, exhibit a twisted ribbon structure reflecting the helicoidal arrangement of the horizontal chitin-protein microfibrils.     

Ontogeny of salinity tolerance and hyper-osmoregulation by embryos of the intertidal crabs Hemigrapsus edwardsii and Hemigrapsus crenulatus (Decapoda, Grapsidae): survival of acute hyposaline exposure

The adults of Hemigrapsus edwardsii and Hemigrapsus crenulatus are euryhaline crabs and strong hyper-osmoregulators. Their embryos are carried externally attached to the abdominal pleopods of female crabs, where they are exposed to temporal and spatial changes in salinity associated with their intertidal and estuarine habitats. Although embryos lack the branchial and excretory organs responsible for adult osmoregulation, post-gastrula embryos were highly tolerant of exposure to hypo-osmotic sea water. Detached eggs (embryos+envelopes), of both species, at all developmental stages between gastrulation and hatching, exhibited 80-100% survival for periods up to 96 h in sea water (osmolality, 1050 mmol kg(-1)) and in dilutions to 50%, 10%, and 1%. Cleavage stages were less tolerant of dilution; H. edwardsii, <50% survived 24 h in 10% sea water; H. crenulatus <50% survived 6 h in 10% sea water. Post-gastrulation stages strongly hyper-osmoregulated but cleavage stages were hyper-osmoconformers (maintaining internal osmolality approximately 150 mmol kg(-1) above external). Osmoregulatory capacity was reduced just prior hatching, particularly in H. crenulatus, although salinity tolerance remained high. Gastrulation therefore marks a critical stage in the ontogeny of osmoregulation and salinity tolerance. Total Na+/K(+)-ATPase activity increased greatly during embryogenesis of H. crenulatus (undetectable in blastulae; gastrulae 0.31+/-0.05 pmol P(i) embryo(-1) min(-1); pre-hatching 16.4+/-1.0 pmol P(i) embryo(-1) min(-1)). Na+/K(+)-ATPase activity increased in embryos exposed to dilute sea water for 24 h implicating regulation of this transporter in a short-term acclimation response.     

The cuticle of the crabs Cancer pagurus L. and Carcinus maenas (L.)

Fine closely-packed parallel fibres pass obliquely, at an angle of about 10° to the horizontal, through the cuticles of Cancer and Carcinus . They lie on axes parallel to the four faces of an obtuse pyramid, and have no counterpart in the model proposed by Bouligand (1965, 1971, 1972).
Replication of vertically broken surfaces of cuticle on cellulose acetate film shows that the laminae of the cuticle are discrete structural entities which preserve their identity around the angle formed by two vertical faces meeting at right-angles. This does not conform to the requirements of the Bouligand model.     

Ionoregulatory and urinary responses to emersion in the mangrove crab Ucides cordatus and the intertidal crab Carcinus maenas

The mangrove crab Ucides cordatus showed a significant reduction in its output of isosmotic urine (to 60% of submerged rates) in humid, emersed conditions when in apparent water balance. A similar, but lesser, reduction was observed in the intertidal temperature species Carcinus maenas. Some of the decrease is effected by increased water withdrawal of an isosmotic fluid in the antennal gland (inulin urine to haemolymph concentration ratio=1.27) in Ucides but most is due to the reduced filtration rate which decreases from 3.41 to 2.19 ml·100 g^-1·day^-1 (in Carcinus from 7.37 to 5.88). In Carcinus, inulin urine to haemolymph concentration ratios are within the range 1.34–1.41 and show significant change on emersion. Comparisons of ion and inulin clearance ratios revealed interspecific differences in renal handling of Na⁺ and Cl^-. During emersion total Na⁺ efflux is markedly reduced in both species. Urinary Na⁺ losses are responsible for most of the efflux in Carcinus but represent only 50% of the total in Ucides. Significant Na⁺ uptake from interstitial water occurs in Ucides. REductions in ion and water loss appear to be mainly dependent on decreases in urine output (clearance rate). However, some released urine was found to accumulate in the branchial chambers in both species. Whether or not extrarenal reabsorption of ions from released urine occurs in the gill chambers in either crab species remains to be proved.Abbreviations CPM counts per minute - ECF extracellular fluid - k rate constant for inulin clearance - k ^Na rate constant for sodium efflux - J _out ^Na sodium efflux rate - Q _tot total radioactivity injected - SEM standard error of mean - SW sea water - 9-SW sea water, salinity 9 mg·ml^-1 (ppt) - 26-SW sea water, salinity 26 mg·ml^-1 (ppt) - U/B ratio of urine (U) to haemolymph (B) concentrations - V _c1 clearance rate - V _u urine output - V _a inulin space     

The ultrastructure of the sinus gland of Carcinus maenas (Crustacea: Decapoda)

Summary The sinus gland of Carcinus maenas consists of the swollen axonal endings of the neurosecretory cells of the major ganglia and acts as a storage release centre for the membrane bound neurosecretory material. These neurosecretory granules fall into five different types based on size and electron density. Their contents are released by exocytosis of the primary granules or smaller units budded from the primary granules.I thank Professor E. Naylor for his constant advice and Professor E. W. Knight-Jones, Department of Zoology, University College, Swansea, for the provision of laboratory facilities. I am grateful to the Science Research Council for the financial support. Finally, I thank the Electron Microscope Unit, Southampton General Hospital, where the work was completed.     

Comparative morphology of larvae of coastal crabs (Crustacea: Decapoda: Varunidae)

The larval development of three crabs of the Varunidae family, (Hemigrapsus sanguineus, H. penicillatus, and H. longitarsis), widely spread in Russian waters of the Sea of Japan, were studied under laboratory conditions. At a temperature of 20–22°C and a salinity of 32‰ about 30% of larvae a complete developmental cycle, including five zoeal stages and megalopa, took from 22 to 30 days. All larval stages are illustrated and described in detail. Zoea I and zoea II of the studied crabs are not distinguishable. Zoea III–V of these species differ in the number of dorsomedial setae on the abdominal somite I and in the number of setae on the posterodorsal arch. The megalopae of three Hemigrapsus species possess a different number of segments, aesthetascs and setae on the antennular exopod. In spite of the great similarity of larvae of genera Hemigrapsus and Eriocheir, the latter possesses a number of distinctive features in all developmental stages, supporting the separation of these genera.     

An ultrastructural investigation of the hypocerebral organ of the adult Euperipatoides kanangrensis (Onychophora, Peripatopsidae)

The hypocerebral organs of Euperipatoides kanangrensis are a pair of spherical vesicles located ventral to the cerebral ganglia. They develop in the embryo from the most anterior pair of ventral organs, in the antennal segment. The wall of each hypocerebral organ is a dense epithelium of elongate cells with peripheral nuclei. The cytoplasm of the cells includes numerous mitochondria, Golgi bodies and microtubules. The small lumen, located eccentrically within the organ, contains concentrically layered electron-dense material resembling cuticle.Each hypocerebral organ is enclosed by a layer of extracellular matrix continuous with that surrounding the adjacent cerebral ganglion. There are no nerve connections between ganglion and organ, but cellular connections traverse the intervening matrix and could serve as a communication pathway. The ultrastructure of the hypocerebral organs indicates that they are glands.     

Larval abundance and recruitment of Carcinus maenas L. close to its southern geographic limit: a case of match and mismatch

In the Ria Formosa lagoon (S. Portugal), Carcinus maenas larvae have been observed only during the cold part of the year (October–May), when the water temperature stays below 23 °C. Larvae suffer high mortality during day time (about 80% during 6 h). Hence, they will only develop outside the lagoon, as proved by the observation of zoea I stages exclusively. Main recruitment starts in April, i.e. much later than the first larvae appear. Small crabs prefer Zostera noltii patches and colonize other sites when the carapace width exceeds about 5 mm. A reserve of some small crabs is found in Zostera noltii patches nearly all year round. It is postulated that one of the key factors for successful propagation of the species is a match between physiological reactions (here in particular temperature effect on reproductive cycle and larval release), and the ecological conditions for larval survival and recruitment, such as predator impact, food availability or currents.     

Electron microscopic study of the mandibular glands of Kalotermes flavicollis Fabr. (Isoptera; Calotermitidae)

Summary The mandibular glands of Kalotermes were examined in different castes. They showed sexual dimorphism in the soldiers and primary reproductives. Moreover, in female soldiers and queens, mandibular gland cells contained numerous crystalline structures of mitochondrial origin. The role of these glands (secretion of saliva or pheromone) is discussed.

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Résumé Les glandes mandibulaires de Calotermes sont étudiées dans les différentes castes. Elles présentent un dimorphisme sexuel chez les soldats et les sexués. Après la mue imaginale, les cellules glandulaires ont toujours une activité sécrétrice. De plus, chez les soldats femelles et les reines, elles contiennent de nombreuses formations cristallines d'origine mitochondriale. Le rôle de ces glandes (sécrétion de salive ou de phéromone) est discuté.

     

Synapse-like contacts between axons of the pineal tract and the subcommissural organ in Rana perezi (Anra) and their absence in Carassius auratus (Teleostei): ultrastructural tracer studies

The present study was designed to investigate the controversial subject of the existence of a neural input from the pineal organ via the pineal tract to the subcommissural organ (SCO) in teleosts and anurans. Horseradish peroxidase was injected into the pineal organ and pineal tract of Carassius auratus and Rana perezi. Within the pinealofugal fibers the tracer was visualized at the light-and electron-microscopic levels either by immunocytochemistry using an anti-peroxidase serum, or by revealing the enzymatic activity of peroxidase. In both species, labeled myelinated and unmyelinated fibers of the pineal tract were readily traced by means of electron microscopy. In R. perezi, numerous terminals contacting the SCO cells in a synapse-like (synaptoid, hemisynaptic) manner bore the label, whereas a different population of endings was devoid of the tracer, indicating that in this species the SCO receives a dual neural input, one of pineal origin, the other of unknown source and nature. In the SCO of C. auratus, neither labeled nor unlabeled synapse-like contacts were found. Thus, in this latter species, a direct neural input to the SCO is missing. It is concluded that the secretory activity of the SCO can be controlled by different mechanisms in different species, and that more than one neural input mechanism may operate in the same species.     

Localization of pigment-dispersing hormone (PDH) immunoreactivity in the central nervous system of Carcinus maenas and Orconectes limosus (Crustacea), with reference to FMRFamide immunoreactivity in O. limosus

Summary By use of antisera raised against synthetic pigment-dispersing hormone (PDH) of Uca pugilator and FMRFamide, the distribution of immunoreactive structures in the central nervous system (CNS) of Carcinus maenas and Orconectes limosus was studied by light microscopy. In both species, a total of 10–12 PDH-positive perikarya occur amongst the anterior medial, dorsal lateral and angular somata of the cerebral ganglion (CG). In C. maenas, one PDH-perikaryon was found in each commissural ganglion (COG) and several more in the thoracic ganglion. In O. limosus, only four immunopositive perikarya could be demonstrated in the ventral nerve cord, i.e., two somata in the anterior and two in the posterior region of the suboesophageal ganglion (SOG). PDH-immunoreactive tracts and fiber plexuses were present in all central ganglia of both species, and individual axons were observed in the connectives. FMRFamide-immunoreactivity was studied in O. limosus only. Neurons of different morphological types were found throughout the entire CNS, including numerous perikarya in the anterior medial, anterior olfactory, dorsal lateral and posterior cell groups of the CG. Four perikarya were found in the COG, six large and numerous smaller ones in the SOG, and up to eight cells in each of the thoracic and abdominal ganglia. In each ganglion, the perikarya form fiber plexuses. Axons from neurons belonging to the CG could be traced into the ventral nerve cord; nerve fibers arising from perikarya in the SOG appeared to project to the posterior ganglia. In none of the structures examined colocalization of PDH- and FMRF-amide-immunoreactivity was observed.Dedicated to Prof. K.-E. Wohlfarth-Bottermann on the occasion of his 65th birthday     

Ultrastructure of Haller's organ in the tick Amblyomma americanum (L.)

Summary Haller's organ on the tarsus of the tick Amblyomma americanum (L.) (Acarina: Ixodidae; nymphal stage) was studied by scanning and transmission electron microscopy. It consists of a distal bristle group, (the anterior pit), and a proximal capsule which encloses several sensilla. The seven sensilla of the anterior pit (A1–A7) are all thick-walled and multi-innervated (2–9 neurons), but at least three different types can be differentiated. Sensilla A1 and A2 possess large, plugged pores (>1000 Å) and are the only sensilla with branching dendrites. A3 and A5 are characterized by a spoke-wheel arrangement of the cuticle wall and very fine pores (100–200 Å) penetrating the spokes centrally; A4, A6, and A7 do not exhibit any pore system but a single opening at the bristle tip is assumed.The capsule contains seven thin-walled, blunt-tipped sensilla, and several non-sensory cuticular projections (pleomorphs). All of these sensilla possess large plugged pores in the cuticle wall and numerous dendritic branches of several neurons (3–5) in the lumen. Glandular openings were found inside the capsule; their significance is discussed.The fine structure of Haller's organ supports the functions postulated by Lees (1948), namely olfaction for the capsule and humidity reception (among others) for the anterior pit.This research was supported in part by the Office of Naval Research, and by NIH Training grant ES 00069. Paper no. 3459 of the Journal Series of the North Carolina State University Agricultural Experiment Station, Raleigh.     

Organization of a phyllobranchiate gill from the green shore crab Carcinus maenas (Crustacea,Decapoda)

Summary The phyllobranchiate gills of the green shore crab Carcinus maenas have been examined histologically and ultrastructurally. Each gill lamella is bounded by a chitinous cuticle. The apical surface of the branchial epithelium contacts this cuticle, and a basal lamina segregates the epithelium from an intralamellar hemocoel. In animals acclimated to normal sea water, five epithelial cell types can be identified in the lamellae of the posterior gills: chief cells, striated cells, pillar cells, nephrocytes, and glycocytes. Chief cells are the predominant cells in the branchial epithelium. They are squamous or low cuboidal and likely play a role in respiration. Striated cells, which are probably involved in ionoregulation, are also squamous or low cuboidal. Basal folds of the striated cells contain mitochondria and interdigitate with the bodies and processes of adjacent cells. Pillar cells span the hemocoel to link the proximal and distal sides of a lamella. Nephrocytes are large, spherical cells with voluminous vacuoles. They are rimmed by foot processes or pedicels and frequently associate with the pillar cells. Glycocytes are pleomorphic cells packed with glycogen granules and multigranular rosettes. The glycocytes often mingle with the nephrocytes. Inclusion of the nephrocytes and glycocytes as members of the branchial epithelium is justified by their participation in intercellular junctions and their position internal to the epithelial basal lamina.     

The Use of Artificial Benthic Collectors for Assessment of Spatial Patterns of Settlement of Megalopae of Carcinus maenas (L.) (Brachyura: Portunidae) in the Lower Mira Estuary, Portugal

Artificial benthic collectors have been widely used for the assessment of settlement rates of decapod crustaceans. However, to date no consistent works have addressed spatial patterns of settlement in different estuarine habitats, and no specific studies targeted the interaction of artificial surfaces with the surrounding natural substrate. It may be expected that the artificial surface may produce a different thigmotactic response when compared to the natural substrate, which may limit the use of this technique for assessment of natural settlement rates. In this study the settlement rates of megalopae of the estuarine crab Carcinus maenas were addressed, specifically deploying artificial benthic collectors in different habitats both intertidal and subtidal in the lower Mira estuary. A number of experiments were performed concerning stratification and temporal fluctuations of settlement. Further, the interaction of collector surface with the surrounding substrate was investigated, by comparing settlement rates in natural and artificial substrates in different habitats. Results have shown significant differences in settlement between different estuarine habitats, both in spatially replicated experiments and in a high-resolution temporal experiment. However, comparison between settlement rates in artificial and natural substrates has shown that there is a strong interference between collectors and surrounding substrate, limiting interpretation of results concerning settlement rates in artificial substrate alone.