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PubMed1991

Histamine: a putative afferent neurotransmitter in Limulus eyes.

Histamine has been proposed as a photoreceptor neurotransmitter in two major groups of arthropods, the insects and the crustacea. In this study biochemical and immunocytochemical approaches were used to examine the synthesis, endogenous content, and cellular distribution of histamine in the visual system of the horseshoe crab Limulus polyphemus, an ancient chelicerate arthropod. Studies with this animal have been critical to our understanding of the basic processes of vision. High-voltage paper electrophoresis was used to assay for histamine synthesis in Limulus tissues incubated with radiolabeled histidine; histamine synthesis was detected in the lateral, median, and ventral eyes and optic nerves and in the visual centers in the brain. Endogenous histamine, assayed as its orthophthalaldehyde derivative by high-performance liquid chromatography and electrochemical detection, was also detected in these tissues. Immunocytochemical analyses, with an antiserum directed against a protein conjugate of histamine, revealed histamine-like immunoreactivity in the somata of photoreceptors in each of the eyes and in the regions of the brain where the photoreceptors terminate. Histamine-like immunoreactivity was also intense in the cell bodies and axon collaterals of eccentric cells in the lateral eye and in eccentric cell projections in the brain. These results show that histamine is a major biogenic amine in the Limulus visual system, and they suggest that this amine is involved in transmitting visual information from the eyes to the brain and in lateral inhibition, a fundamental mechanism for processing visual information in the lateral eye.

Afferent Pathways↗
PubMed2005

Stages of embryonic development in the amphipod crustacean, Parhyale hawaiensis.

Studying the relationship between development and evolution and its role in the generation of biological diversity has been reinvigorated by new techniques in genetics and molecular biology. However, exploiting these techniques to examine the evolution of development requires that a great deal of detail be known regarding the embryonic development of multiple species studied in a phylogenetic context. Crustaceans are an enormously successful group of arthropods and extant species demonstrate a wide diversity of morphologies and life histories. One of the most speciose orders within the Crustacea is the Amphipoda. The embryonic development of a new crustacean model system, the amphipod Parhyale hawaiensis, is described in a series of discrete stages easily identified by examination of living animals and the use of commonly available molecular markers on fixed specimens. Complete embryogenesis occurs in 250 h at 26 degrees C and has been divided into 30 stages. This staging data will facilitate comparative analyses of embryonic development among crustaceans in particular, as well as between different arthropod groups. In addition, several aspects of Parhyale embryonic development make this species particularly suitable for a broad range of experimental manipulations.

Amphipoda↗
PubMed2004

Improvement of microanatomical research by combining corrosion casts with MicroCT and 3D reconstruction, exemplified in the circulatory organs of the woodlouse.

A SkyScan-1072 high-resolution desktop microtomograph was used to visualize the 3D morphology of the circulatory system of Porcellio scaber (Crustacea, Malacostraca, Isopoda) at the micrometer level. Prior to MicroCT investigation, the circulatory system was injected with a fast-hardening resin, permitting a better contrast of the vascular structures that are normally hollow after fixation. Stacks of tomographic images were used to reconstruct the circulatory system three-dimensionally. The spatial resolution achieved by the microtomograph is about 3.5 microm. The technique provides detailed tomographic images of the inner organs and makes precise 3D reconstructions of the circulatory system in arthropods possible. The results were compared with conventional histological data. MicroCT permits a high number of specimens to be screened in a short time, which is essential for the improvement of modern morphological research in a cladistic framework. Histological techniques, however, are still essential for gathering information at the cellular level.

Acrylic Resins↗
PubMed1975

Effect of limb regeneration on size increase at molt of the shore crabs Hemigrapsus oregonensis and Pachygrapsus crassipes.

Size increase at molt is reduced following multiple limb regeneration in the shore crabs, Hemigrapsus oregonensis and Pachygrapsus crassipes. Limb loss per se does not influence postmolt size. Effect of increasing number of regenerating limbs is additive. Postmolt size is programmed early in the premolt period of the preceding instar and is probably not readily influenced by water uptake mechanics at ecdysis. A simple model for growth, molting, and regeneration in heavily calcified Crustacea is developed from the viewpoint of adaptive strategies and energetic considerations.

Animals↗
PubMed1995

Proteins of crustacean exoskeleton: IV. Partial amino acid sequences of exoskeletal proteins from the Bermuda land crab, Gecarcinus lateralis, and comparisons to certain insect proteins.

As in all decapod Crustacea, the exoskeleton of the land crab Gecarcinus lateralis consists of four layers. Prior electrophoretic analysis of proteins extracted from these layers revealed an abundance of small M(r) proteins with acidic pIs are found in insect cuticle (O'Brien et al. [1991 Biol. Bull., 181:427-441). Further, immunological cross-reactivity between crab exoskeletal proteins and insect cuticular proteins has been demonstrated (Kumari and Skinner [1993] J. Exp. Zool., 265:195-210). Partial amino acid sequences of a number of proteins from the four exoskeletal layers are described here. Proteins were electrophoresed on two-dimensional (2D) gels, transferred to polyvinylidene difluoride (PVDF) membranes, and stained; individual spots were recovered and their N-termini were sequenced. In addition, a 14-kDa protein (pI = 5.4) from membranous layer (ML14) was eluted from 2D gels and digested with endoproteinase Lys-C; N-termini of its constituent peptides were sequenced. The two epicuticular proteins differed from each other. Three proteins with identical electrophoretic mobility isolated from exocuticle, endocuticle, and membranous layer appeared to have identical N termini, while another electrophoretically identical set from the three layers appeared identical with each other but differed in three positions from the first set. Two proteins from the membranous layer both had a mass of 25 kDa but different isoelectric points. Their sequences were indistinguishable from each other but clearly distinct from another membranous layer protein. Another distinct sequence was found in a 14-kDa protein from endocuticle, while a less acidic pair of 14-kDa proteins from endocuticle and membranous layer were quite similar to one another. The three internal peptide fragments from ML14 were distinct, but one had regions similar to the ML14 N terminus. One crab exoskeletal protein sequence was similar to some structural proteins of vertebrates, whereas others had motifs found in insect cuticular proteins. The sequence similarities identified did not account for the antibody cross-reactivity.

Amino Acid Sequence↗
PubMed2005

Ontogeny of the Marmorkrebs (marbled crayfish): a parthenogenetic crayfish with unknown origin and phylogenetic position.

Development, growth, and egg production of the Marmorkrebs (marbled crayfish), a crayfish with parthenogenetic reproduction, uncertain geographic origin, and taxonomic position, was studied under laboratory conditions. Length and weight increments strongly depended on temperature being highest at 30 degrees C, and lowest at 15 degrees C. At 25 degrees C, cephalothorax length and weight increased by 17.5 mm and 1700 mg, respectively, in the course of 150 d, whereas at 15 degrees C these parameters increased by only 7 mm and 100 mg during the same period of time. Photoperiod slightly affected growth at 25 degrees C. During growth experiments, mortality was lower at 20 degrees C compared to higher (25 degrees , 30 degrees C) or lower temperatures (15 degrees C), and lower under short-day than under long-day conditions. Females matured early (at an age of 141-255 d, a cephalothorax length of 14-21.5 mm, and a weight of 0.63-2 g) compared to other crayfish species. Reproductive females with a cephalothorax length of between 25-35 mm produced large clutches (up to 416 eggs) and brooding periods varied between 22 and 42 d. In order to establish a staging scheme for Marmorkrebs embryos, embryos were photographed, externally visible ontogenetic events charted, and dissected embryos stained with a nuclear dye. These experiments indicate that their development is virtually identical to that of other crayfish. In conclusion, these results and others show that the Marmorkrebs may be taken as a representative valid model organism for future developmental studies on Crustacea.

Acclimatization↗
PubMed2004

Molecular characterization of a cDNA encoding vitellogenin in the coonstriped shrimp, Pandalus hypsinotus and site of vitellogenin mRNA expression.

In order to determine the primary structure of vitellogenin in a protandric species, the coonstriped shrimp Pandalus hypsinotus, we previously purified four vitellin components (designated as VnA, VnB, VnC, and VnD, respectively), and chemically analyzed their partial amino acid sequences. In this study, we subsequently cloned a cDNA encoding vitellogenin in this species based on the N-terminal and internal amino acid sequences of VnA, as well as the N-terminal sequence of VnC. The open reading frame of this cDNA encoded a pro-vitellogenin in which vitellins were arranged as follows: NH2-VnA-VnB-VnC/D-COOH. The deduced amino acid sequence possessed a single consensus cleavage sequence, R-X-K/R-R, along the lines of vitellogenins reported in other crustaceans and insects, and the N-terminal sequence of VnB was immediately preceded by this sequence. The comparison of primary structures revealed the existence of a basic and characteristic structure for the vitellogenin molecule in decapod crustacean species, and phylogenetic analysis reflected the current taxonomic classifications of Crustacea. An approximately 8 kb-long transcript of the vitellogenin gene was detected in the hepatopancreas of female shrimps having a gonadosomatic index higher than 1.0 by Northern blot analysis, but was not observed in the hepatopancreas and gonads of male shrimps and the hepatopancreas of female shrimps having a gonadosomatic index lower than 1.0. These results indicate that the hepatopancreas is responsible for vitellogenin synthesis.

Amino Acid Sequence↗
PubMed2004

Life stages and reproductive components of the Marmorkrebs (marbled crayfish), the first parthenogenetic decapod crustacean.

Recently, we briefly reported on the first case of parthenogenesis in the decapod Crustacea which was found in the Marmorkrebs or marbled crayfish, a cambarid species of unknown geographic origin and species identity. Curiously, this animal is known only from aquarium populations, where it explosively propagates. By means of light and electron microscopic techniques we have now investigated the reproductive components of this crayfish, using more than 100 specimens ranging from hatchling to repeatedly spawned adult. Additionally, we documented its principal life stages. Our results revealed that the external sexual characters and also the gonads of the marbled crayfish are purely female, making this fast-reproducing species a good model for investigating female reproductive features in crayfish. Testicular tissues, ovotestes, or male gonoducts, gonopores, or gonopods were never found, either in small juveniles or large adult specimens, confirming the parthenogenetic nature of this crayfish. Parthenogenesis may have arisen spontaneously or by interspecific hybridization since Wolbachia-like feminizing microorganisms were not found in the ovaries. The external sexual characters of the marbled crayfish are first recognized in Stage 4 juveniles and are structurally complete approximately 2 months after hatching in specimens of approximately 2 cm total length. In the same life stage the ovary is fully differentiated as well, although the oocytes are in previtellogenic and primary vitellogenic stages only. The architecture of the mature ovary and also the synchronous maturation of cohorts of primary vitellogenic oocytes by secondary vitellogenesis are in general agreement with data published on ovaries of bisexual crayfish. New results were obtained with respect to the muscular nature of the ovarian envelope and its extensive proliferation after the first spawning, the distribution of hemal sinuses in the ovarian envelope and in the interstitium around the oogenetic pouches, the high transport activity of the follicle cells, and the colonization of oogenetic pouches by previtellogenic oocytes that originate in the germaria. Investigation of the nuclei of oocytes in the germaria and oogenetic pouches revealed no signs of meiosis, as usually found in females of bisexual decapods, suggesting that parthenogenesis in the marbled crayfish might be an apomictic thelytoky. The detection of new rickettsial and coccidian infections in the ovary and further organs raises fears that the marbled crayfish might endanger native European species by transmission of pathogens once escaped into the wild.

Animals↗
PubMed2002

A new look at an old visual system: structure and development of the compound eyes and optic ganglia of the brine shrimp Artemia salina Linnaeus, 1758 (Branchiopoda, anostraca).

Compared to research carried out on decapod crustaceans, the development of the visual system in representatives of the entomostracan crustaceans is poorly understood. However, the structural evolution of the arthropod visual system is an important topic in the new debate on arthropod relationships, and entomostracan crustaceans play a key role in this discussion. Hence, data on structure and ontogeny of the entomostracan visual system are likely to contribute new aspects to our understanding of arthropod phylogeny. Therefore, we explored the proliferation of neuronal stem cells (in vivo incorporation of bromodeoxyuridine) and the developmental expression of synaptic proteins (immunohistochemistry against synapsins) in the developing optic neuropils of the brine shrimp Artemia salina Linnaeus, 1758 (Crustacea, Entomostraca, Branchiopoda, Anostraca) from hatching to adulthood. The morphology of the adult visual system was examined in serial sections of plastic embedded specimens. Our results indicate that the cellular material that gives rise to the visual system (compound eyes and two optic ganglia) is contributed by the mitotic activity of neuronal stem cells that are arranged in three band-shaped proliferation zones. Synapsin-like immunoreactivity in the lamina ganglionaris and the medulla externa initiated only after the anlagen of the compound eyes had already formed, suggesting that the emergence of the two optic neuropils lags behind the proliferative action of these stem cells. Neurogenesis in A. salina is compared to similar processes in malacostracan crustaceans and possible phylogenetic implications are discussed.

Age Factors↗
PubMed1982

All-or-none control of the bursting properties of the pacemaker neurons of the lobster pyloric pattern generator.

In Crustacea the central pattern generator for the pyloric motor rhythm (filtration to the midgut) is known to be located within the stomatogastric ganglion (STG); its cycling activity is known to be organized by three endogenous burster neurons acting as pacemakers and driving 11 follower neurons. In Homarus, recordings from the isolated stomatogastric nervous system (Fig. 1) indicate that (1) the pyloric output can be generated only when the STG is afferented (i.e., connected to the more rostral oesophageal and commissural ganglia) (Fig. 2) and (2) the deafferentation of the STG results in a complete loss of the bursting properties of the pacemaker neurons (Fig. 4). Manipulation of the STG inputs responsible for unmasking the properties of the pacemakers strongly suggests that (1) they are not phasic inputs (Fig. 5) and (2) they are long-term acting inputs (Fig. 6). These results provide evidence for a neural all-or-none control of the bursting properties of the pacemaker neurons of a motor pattern generator.

Afferent Pathways↗
PubMed1990

Conservation of antigen 3G6: a crystalline cone constituent in the compound eye of arthropods.

A monoclonal antibody (MAb), 3G6, highly selective for neuropil glia in the CNS of the house cricket Acheta domesticus, also demonstrates remarkable selectivity for the nonneuronal crystalline cone cells of the compound eye. MAb 3G6 labels cone cells in eucone eyes throughout Insecta, from ancestral forms such as the bristle tail to the more recent honeybee; eucone structures are also recognized in Crustacea. Analogous nonneural structures found in pseudocone or acone eyes also express detectable 3G6 immunoreactivity. Immunoblot analysis demonstrates that MAb 3G6 binds to similar Mr85 kDa glycoproteins in the cricket CNS and retina, corresponding to the glial and crystalline cone forms of the antigen. Further, polypeptides of similar relative mass are also recognized in the eucone eye of the butterfly Pieris and the pseudocone eye of the fly Calliphora. The properties and function of glycoprotein 3G6 in the CNS and retina are yet to be explored. However, the finding that a unique antigen is highly conserved within the crystalline cone or analogous regions of the retina throughout the Arthropoda lends support at the molecular level to the notion that the arthropod compound eye has a monophyletic origin.

Animals↗
PubMed1994

Regeneration and molting effects on a proprioceptor organ in the Dungeness crab, Cancer magister.

Decapoda Crustacea molt in order to grow; some species, such as the Dungeness crab Cancer magister, achieve a very large size. Does sensory neuron hyperplasia in internal proprioceptors accompany this growth? To determine this, neurons in propodite-dactylopodite (PD) chordotonal organs were counted in first walking legs of juvenile (5th through 9th instar) and adult (10th through 13th instar) C. magister. We found that the PD organs of J5 crabs have about 56 neurons; the number increases to about 61 neurons in J6 crabs. Significant hyperplasia now occurs because an average of 79 neurons are found in the PD organs of J7 crabs. Little to no hyperplasia accompanies the several succeeding juvenile and adult molts (ca. 82-86 neurons are present). Because autotomized limbs are regenerated upon molting, we also examined how the number of PD organ neurons in regenerated legs compares with those of pristine legs. Newly regenerated legs (termed 1st stage regenerates) have fewer sensory neurons than do their contralateral pristine partners (65 vs 81); larger regenerated legs which have attained nearly normal size as a result of additional molts (2nd stage regenerates) still have fewer neurons than their pristine partners (69 vs 81). Additionally, in contrast to those of pristine walking legs, the elastic strand of PD organs from 1st stage regenerates in a misshapen sheet containing a cluster of small neurons with no obvious functional organization. Nonetheless, neurophysiological recordings indicate that all the receptor types typical for pristine legs (movement and position cells) are represented. The PD organs of 2nd stage regenerates differentiate to the shape and neuronal organization of pristine legs.

Animals↗
PubMed2001

Structural and functional properties of hemocyanin from Cyanagraea praedator, a deep-sea hydrothermal vent crab.

Cyanagraea praedator (Crustacea: Decapoda: Brachyura) is an endemic species of the East Pacific Rise hydrothermal vents, living in the upper part of black smoker chimneys. Because we were seeking species that have made respiratory adaptations to the hydrothermal environment, we looked at Cyanograea hemocyanin (Hc) and determined its quaternary structure and the oxygen-binding properties in relation to temperature, pH, and lactate. C. praedator Hc is composed of dodecamers and hexamers, with dodecamers formed by the perpendicular association of two hexamers. The composition of these polymers was determined by electrophoresis and, for the first time, by electrospray mass spectrometry. Dodecamers and hexamers are composed of six subunits common to the two forms, with molecular mass ranging from 75,008 Da to 75,534 Da. In addition, we found two dodecamer-specific subunits, at 75,419 Da and 75,629 Da. The native hemocyanin possesses a high oxygen affinity (P(50) varies between 4 and 10 Torr at pH 7.5, 15 degrees C) and a large Bohr coefficient (Delta log P(50)/DeltapH approximately -1.8). Oxygen affinity is not affected by lactate or, surprisingly, temperature between 5 degrees C and 35 degrees C (DeltaH = 1.16 kJ/mol(1) 5-35 degrees C). Dialysis of native hemolymph elicited a significant increase in Hc-O(2) affinity (DeltaP(50) = 2.5 Torr at pH 7.5), an effect opposite the usual trend observed for crustacean hemocyanins. In this article these functional properties are interpreted in relation to characteristics of the environment.

Adaptation, Physiological↗
PubMed2004

Evidence for occurrence of an organophosphate-resistant type of acetylcholinesterase in strains of sea lice (Lepeophtheirus salmonis Krøyer).

Acetylcholinesterase (AChE) is the target of a major pesticide family, the organophosphates, which were extensively used as control agents of sea lice on farmed salmonids in the early 1990s. From the mid-1990s the organophosphates dichlorvos and azamethiphos were seriously compromised by the development of resistance. AChE insensitive to organophosphate chemotherapeutants has been identified as a major resistance mechanism in numerous arthropod species, and in this study, target-site resistance was confirmed in the crustacean Lepeophtheirus salmonis Krøyer isolated from several fish-farming areas in Norway and Canada. A bimolecular rate assay demonstrated the presence of two AChE enzymes with different sensitivities towards azamethiphos, one that was rapidly inactivated and one that was very slowly inactivated. To our knowledge this is the first report of target-site resistance towards organophosphates in a third class of arthropods, the Crustacea.

Acetylcholinesterase↗
PubMed2006

The use of a test battery in marine ecotoxicology: the acute toxicity of sodium dodecyl sulfate.

For a toxicity assessment of substances entering the marine environment, it is preferable to carry out ecotoxicological tests on a base-set of taxa utilizing target species belonging to different trophic levels. In this study a battery composed of Vibrio fischeri (bacteria), Dunaliella tertiolecta (algae), Tigriopus fulvus (crustacea), Paracentrotus lividus (echinodermata), and Dicentrarchus labrax (pisces) was used for acute toxicity testing of sodium dodecyl sulfate (SDS). SDS is an anionic surfactant widely employed in industry, agriculture, and domestic usage and therefore is found in abundance in the environment, particularly in the sea. The mean values of EC50 obtained were 2.6, 4.8, 7.4, 3.2, 7.3 mg L(-1), respectively, for V. fischeri, D. tertiolecta, T. fulvus, P. lividus, and D. labrax. The results indicate the high acute toxicity of SDS with respect to all the trophic levels represented by the target species. In addition, they highlight the usefulness of employing a base-set of taxa rather than a single species in toxicological tests, in order to obtain more reliable information for the evaluation of toxicity and potential hazards to the marine environment of selected substances.

Aliivibrio fischeri↗
PubMed1998

Active brood care in an amphipod: influences of embryonic development, temperature and oxygen.

Female amphipods (Crustacea) carry their fertilized eggs in an external brood pouch until the fully formed juveniles emerge (passive brood care). They may also direct specific maternal activities towards the brood (active brood care). We show that Crangonyx pseudogracilis, which typically populates fresh-waters subject to wide fluctuations in temperature and dissolved oxygen, engages in a highly responsive form of active brood care. This involves a flexing motion by the female that expands the brood pouch and increases the suspension of the eggs in the surrounding medium, accompanied by ventilation of the brood pouch and the 'cycling' of eggs therein. Females also selectively eject nonviable eggs from their broods. We investigated the expression of this brood care behaviour in relation to intrinsic and extrinsic factors relevant to the development of broods. The time spent by females in this behaviour initially increased as embryos developed, but decreased once advanced embryos began to self-ventilate and to have a heart pulse. In addition, both increased temperature and decreased oxygen concentration resulted in increased levels of brooding behaviour. We thus propose that this behaviour functions to ameliorate the microclimate of the brood pouch and serves the changing metabolic demand of the brood, as influenced by the interaction of embryonic development with temperature/dissolved oxygen regime. In addition, this behaviour may be a key adaptation facilitating the success of this North American species as an invader of disturbed and polluted freshwaters in Europe and elsewhere. Evidence is emerging that other amphipods associated with harsh environmental conditions also show such active maternal brood care. Copyright 1998 The Association for the Study of Animal Behaviour.

Journal Article↗
PubMed1998

The role of calceoli in mate assessment and precopula guarding in Gammarus.

Gammarid crustacea exhibit precopula mate guarding in which a male will carry a potential mate beneath his ventral surface, guarding her for several days until she moults and lays her eggs. I investigated the role of the calceoli (bulb-like structures on the second antennae) in mate assessment and precopula pairing in two Gammarus species by comparing the behaviour of males with and without calceoli. Removal of the calceoli had no effect on the pattern of size-assortative mating, nor did it reduce the number of contacts made per unit time between potential mates. However, males without calceoli reduced their assessment of potential mates, were less likely to guard a suitable female and guarded for a shorter period. These data support the hypothesis that calceoli are used during contact assessment to determine a female's intermoult period and hence suitability for guarding. (c) 1998 The Association for the Study of Animal Behaviour.

Journal Article↗
PubMed1998

Cytochrome P450 enzymes belonging to the CYP4 family from marine invertebrates.

Six new cytochrome P450s assigned to the CYP4 family were identified from marine invertebrates belonging to the arthropod, mollusc, and echinoderm phyla. These are the first reported members of the CYP4 gene family from marine invertebrates, and extends the finding of the CYP4 family of cytochrome P450s to molluscs and echinoderms. Members of each phyla (echinodermata, arthropoda (crustacea), and mollusca) expressed genes belonging to the CYP4C subfamily in their respective digestive tissues. A mollusc, the mussel Mytilus galloprovincialis, expressed a gene belonging to a new CYP4 subfamily, CYP4Y. In Northern blotting experiments with digestive tissues of M. galloprovincialis, the expression of the CYP4Y1 gene was found to be inhibited by increasing concentrations of the hydrocarbon beta-naphthoflavone. Thus, the potential use of marine invertebrate CYP4 genes as biomarkers of xenobiotic exposures may be warranted.

Amino Acid Sequence↗