The effect of serotonin and octopamine on the optokinetic response of the crab Leptograpsus variegatus

A standard optokinetic response of the ipsilateral and contralateral (driven) eyes of the crab Leptograpsus variegatus to a sinusoidally oscillating striped drum was established. Optokinetic responses were then measured of animals that had been treated by introducing serotonin and octopamine into the blood stream via the heart and also into the neural tissue of the optic lobes via a micropipette. Both serotonin and octopamine enhance the optokinetic effect when applied in low doses. Experiments show that serotonin is most likely acting closer to the sensory input in the optokinetic system.     

Sodium currents in the giant axon of the crab Carcinus maenas

Measurements were made of the kinetics and steady-state properties of the sodium conductance changes in the giant axon of the crab Carcinus maenas. The conductance measurements were made in the presence of small concentrations of tetrodotoxin and as much electrical compensation as possible in order to minimize errors caused by the series resistance. After an initial delay of 10-150 microsec, the conductance increase during depolarizing voltage clamp pulses followed the Hodgkin-Huxley kinetics. Values of the time constant for the activation of the sodium conductance lay on a bell-shaped curve with a maximum under 180 microsec at -40 mV (at 18 degrees C). Values of the time constant for the inactivation of the sodium conductance were also fitted using a bell-shaped curve with a maximum under 7 msec at -70 mV. The effects of membrane potential on the fraction of Na channels available for activation studied using double pulse protocols suggest that hyperpolarizing potentials more negative than -100 mV lock a fraction of the Na channels in a closed conformation.     

Potassium currents in the giant axon of the crab Carcinus maenas

Measurements were made of the kinetic and steady-state characteristics of the potassium conductance in the giant axon of the crab Carcinus maenas. These measurements were made in the presence of tetrodotoxin, using the feedback amplifier concept introduced by Dodge and Frankenhaeuser (J. Physiol, (London) 143:76-90). The conductance increase during depolarizing voltage-clamp pulses was analyzed assuming that two separate potassium channels exist in these axons. The first potassium channel exhibited activation and fast inactivation gating which could be fitted using the m3h, Hodgkin-Huxley formalism. The second potassium channel exhibited the standard n4 Hodgkin-Huxley kinetics. These two postulated channels are blocked by internal application of caesium, tetraethylammonium and sodium ions. External application of 4 amino-pyridine also blocks these channels.     

Entrainment of bimodal circatidal rhythms in the shore crab Carcinus maenas

Individuals of the shore crab Carcinus maenas were exposed to artificial cycles, applied in tidal antiphase, of pairs of the three major environmental variables that entrain circatidal rhythmicity in this species: salinity, temperature, and hydrostatic pressure. During entrainment, the observed locomotor activity patterns were dominated by exogenous responses to high pressure, low temperature, or low salinity. In subsequent constant conditions, many of the crabs showed bimodal circatidal rhythms, with peaks phased to the times of expected high-tide characteristics of high pressure, low temperature, or high salinity. Similar bimodal rhythms were induced by exposing freshly captured crabs, with free-running circatidal rhythms, to tidal antiphase cycles of each of the three environmental variables applied individually. The hypothesis that circatidal rhythmicity in this species is controlled by at least two separate circatidal oscillators, with differential sensitivities to specific cyclical environmental variables, is discussed.     

Sex-specific mediation of foraging in the shore crab, Carcinus maenas

Experiments were conducted to investigate the sex-specific differences to feeding responses of the shore crab Carcinus maenas throughout the year. Results demonstrate that female shore crabs exhibit stronger feeding responses than males throughout the year with a significantly reduced feeding response in males during the summer months' reproductive season. We also studied the possible function(s) of the moulting hormone, 20-hydroxyecdysone (Crustecdysone) that has been described as a potential female-produced sex pheromone to initiate male reproductive behaviour in a number of crustaceans. We recently presented evidence that for shore crabs this is not the case and now show that the steroid is instead functioning as a sex-specific feeding deterrent protecting the moulting 'soft' female crabs. Whilst male shore crabs were deterred from prey (Mytilus edulis) and synthetic feeding stimulants glycine and taurine when these feeding stimulants were spiked with crustecdysone, intermoult female crabs were significantly less affected and rarely deterred from feeding. This sex specificity of the moulting hormone, in combination with the female sex pheromone, which has no anti-feeding properties, ensures that male crabs mate with soft-shelled, moulted females rather than engage in cannibalism, such as found frequently in cases when soft-shelled females are exposed to intermoult females.     

Cryptic invasions of the crab Carcinus detected by molecular phylogeography

Coastal marine ecosystems world-wide are threatened by invasions of nonindigenous species. The ubiquity of marine sibling species identifiable only by genetic analysis suggests that many invasions are cryptic and therefore undetected, causing an underestimation of the actual number and impacts of invading species. We test this hypothesis with European crabs in the genus Carcinus that have invaded five regions globally. Partial 16S ribosomal RNA gene sequences confirm sibling species status of morphologically similar Atlantic C. maenas and Mediterranean C. aestuarii . Based on 16S rRNA haplotypes, crabs from California, New England and Tasmania were all C. maenas . However, we report the cryptic multiple invasion of both species in Japan and South Africa, where only C. aestuarii and C. maenas , respectively, were previously recognized.     

Ecdysteroid metabolism in a crab: Carcinus maenas L

Ponasterone A (25-deoxy-20-hydroxyecdysone) and 20-hydroxyecdysone were the major ecdysteroids detected in crab hemolymph, although some ecdysone was also present. The metabolism of ponasterone A was examined in intermolt and premolt crabs either by injecting the radiolabeled hormone or by incubating tissues in its presence. Metabolites were extracted from the surrounding seawater and from tissues and separated by high-performance liquid chromatography. Ponasterone A metabolism proceeds through (1) C-25 and C-26 hydroxylation, followed by formation of inactivation products via oxidation of the terminal alcoholic group to a carboxylic residue, (2) conjugation, (3) "binding" to very polar compounds and (4) side-chain scission. The conversion of ponasterone A into 20-hydroxyecdysone, inokosterone (25-deoxy-20, 26-dihydroxyecdysone), 20, 26-dihydroxyecdysone and ecdysonoic acids, as well as the formation of conjugates and of very polar compounds, occurs in various tissues. These metabolites were excreted by both intermolt and premolt crabs.     

Carapace repair in the green crab,Carcinus maenas (L.)

Formation of a circular hole 8–10 mm in diameter in the calcified layers of the carapace from crabs in stage C₄ of the molt cycle stimulates the tissue under and adjacent to the injury to deposit a unique calcified cuticular material below the intact membranous layer. Deposition was followed for 69 days using light microscopic histology, histochemistry, and scanning electron microscopy. Quantitative analyses of CaCO₃ were conducted using atomic absorption spectrophotometry and Gran titration. Spatial distribution of CaCO₃ was determined with X-radiography. A scab is formed by day two under the injury. At four days the epithelium changes from squamous to columnar and deposits a PAS-positive layer with an irregular lamellar fine structure, followed by highly organized lamellae structurally similar to normal exocuticle. Histochemically, however, these lamellae resemble normal endocuticle. CaCO₃ is evident external to the outermost lamellae by day eleven as a fused mass of aragonite granules. The lamellar region calcifies proximally from the outer surface and is amorphous CaCO₃. Repair cuticle is approximately 20%CaCO₃ by weight.     

Characterization of microsatellite loci in the European green crab (Carcinus maenas)

Carcinus maenas (Decapoda: Portunidae) has proven a highly successful invasive marine species whose potential economic and ecological impacts are of great concern worldwide. Here, we characterize 14 polymorphic microsatellite loci in C. maenas and its sister species Carcinus aestuarii. These markers will prove useful for fine‐scale genetic analyses of native and introduced populations, for assessment of the sources and routes of invasion and for evaluation of post‐invasion population dynamics.     

A membrane-bound hemoglobin from gills of the green shore crab Carcinus maenas

Most hemoglobins serve for the transport or storage of O(2). Although hemoglobins are widespread in "entomostracan" Crustacea, malacostracans harbor the copper-containing hemocyanin in their hemolymph. Usually, only one type of respiratory protein occurs within a single species. Here, we report the identification of a hemoglobin of the shore crab Carcinus maenas (Malacostraca, Brachyura). In contrast to the dodecameric hemocyanin of this species, C. maenas hemoglobin does not reside in the hemolymph but is restricted to the gills. Immunofluorescence studies and cell fractioning showed that C. maenas hemoglobin resides in the membrane of the chief cells of the gill. To the best of our knowledge, this is the first time that a membrane-bound hemoglobin has been identified in eukaryotes. Bioinformatic evaluation suggests that C. maenas hemoglobin is anchored in the membrane by N-myristoylation. Recombinant C. maenas hemoglobin has a hexacoordinate binding scheme at the Fe(2+) and an oxygen affinity of P(50) = 0.5 Torr. A rapid autoxidation rate precludes a function as oxygen carrier. We rather speculate that, analogous to prokaryotic membrane-globins, C. maenas hemoglobin carries out enzymatic functions to protect the lipids in cell membrane from reactive oxygen species. Sequence comparisons and phylogenetic studies suggested that the ancestral arthropod hemoglobin was most likely an N-myristoylated protein that did not have an O(2) supply function. True respiratory hemoglobins of arthropods, however, evolved independently in chironomid midges and branchiopod crustaceans.