Aurelin, a novel antimicrobial peptide from jellyfish Aurelia aurita with structural features of defensins and channel-blocking toxins

A novel 40-residue antimicrobial peptide, aurelin, exhibiting activity against Gram-positive and Gram-negative bacteria, was purified from the mesoglea of a scyphoid jellyfish Aurelia aurita by preparative gel electrophoresis and RP-HPLC. Molecular mass (4296.95 Da) and complete amino acid sequence of aurelin (AACSDRAHGHICESFKSFCKDSGRNGVKLRANCKKTCGLC) were determined. Aurelin has six cysteines forming three disulfide bonds. The total RNA was isolated from the jellyfish mesoglea, RT-PCR and cloning were performed, and cDNA was sequenced. A 84-residue preproaurelin contains a putative signal peptide (22 amino acids) and a propiece of the same size (22 amino acids). Aurelin has no structural homology with any previously identified antimicrobial peptides but reveals partial similarity both with defensins and K+ channel-blocking toxins of sea anemones and belongs to ShKT domain family.     

A method for in situ estimation of prey selectivity and predation rate in large plankton, exemplified with the jellyfish Aurelia aurita (L.)

A method to estimate predation rates of large predatory zooplankton, such as jellyfish and ctenophores, is outlined. Large plankton size allows direct visual tracking of the predator during the process of foraging. The presented method is novel in the sense that it measures predation rate of a specific individual plankton predator in situ.After prey has been evacuated from the gut of an individual predator, the predator is incubated in situ, and observed by SCUBA-divers who recapture the individual after a defined time. Given that this incubation time is shorter than prey digestion time, predation rate can be calculated as increase in gut content over time. Clearance rates for different prey can be calculated from predation rates and prey concentrations in the water, allowing accurate estimates of prey selectivity. Thus, the problem of unknown feeding history and feeding environment, which can otherwise be a problem in prey selectivity studies of in situ-captured predators, is circumvented. Benefits and limitations of the method are discussed.The method was applied to adult medusae of the common jellyfish Aurelia aurita. A large variation in number of captured prey was detected both among individual jellyfish and among the various oral arms and gastric pouches within individuals. Clearance rates varied strongly with prey type. The medusae selected large crustacean prey (cladocerans and copepods/copepodites) over echinoderm larvae and copepod nauplii. Prey distribution within the medusae indicates that both tentacles and oral arms were used as prey capturing sites. Food passage time from prey capturing organs to gastric pouches was estimated.     

Plate in the zone of oocyte and germinal epithelium contact in Scyphomedusa Aurelia aurita binds antibodies to ZP-domain protein mesoglein

The morphological features of oocyte and germinal epithelium (epithelial wall of germinal sinus) contact area in Scyphomedusa Aurelia aurita are described. Growing oocytes were divided into seven stages based on oocyte size. The structure revealed the area of contact between the oocyte and the germinal epithelium called the contact plate. During oocyte growth, single granules are fused into the homogeneous mass area of oocyte contact with epithelium. Plate components bound antibodies to mesoglein. It was assumed that the plate material contained ZP-domain proteins. Electrophoresis and immunoblotting results show that proteins immunologically similar to mesoglein have higher molecular masses, probably due to post-translation modifications, which are common for extracellular proteins. On the other hand, however, gonad proteins may be other representatives of jellyfish ZP-domain proteins. Further experiments should be conducted to clarify which alternative is true.     

An investigation of the Common Jellyfish,Aurelia aurita,in the harbour of Trabzon

Summary

Some allometric relationships of the Common Jellyfish, Aurelia aurita, a common species along the Turkish Eastern Black Sea coast, were studied in the harbour of Trabzon. Log bell diameter was linearly related to log wet weight: wet weight = 0.61 . diameter^1.88. Some physico-chemical parameters of the sea water in the harbour are also given.     

In situ estimation of ephyrae liberated from polyps of Aurelia aurita using settling plates in Tokyo Bay, Japan

The continuous changes in the number of newly established polyps of Aurelia aurita (L.) on settling plates under natural conditions were observed from August 1998 to September 1999 in Tokyo Bay, Japan. A sharp decline in survivorship of newly settled polyps was observed within the first few days, however, survivorship of polyps settled in October increased by budding up to 399% after two months. The number of discs in each strobila varied from 1 to 6, however, most of the strobilae formed single discs. The percentage ratios of the total number of ephyrae to the initial number of polyps on settling plates were generally lower than 10%, but the highest ratio of 594.4% was estimated for the polyps settled in October. It is considered that most of the liberated ephyrae originate from the polyps settled in October in Tokyo Bay. This study suggests that the occurrence of ripe medusae with planula larvae throughout the year contributes to the success of settlement and growth of the polyp stage in Tokyo Bay.     

Mesogleal cells of the jellyfish Aurelia aurita are involved in the formation of mesogleal fibres

The extracellular matrix of the jellyfish Aurelia aurita (Scyphozoa, Cnidaria), known as the mesoglea, is populated by numerous mesogleal cells (Mc). We determined the pattern of the Mc and the mesoglea, raised polyclonal antibodies (RA47) against the major mesogleal protein pA47 (47 kDa) and checked their specificity. In the mesoglea, RA47 stains pA47 itself. In immunoblots of Mc, RA47 stains bands of 120 kDa and 80 kDa; weaker staining is observed at pA47. The same staining pattern is seen on blots of jellyfish epidermal cells and of whole Hydra (Hydrozoa) or isolated mesoglea of Hydra. Our data indicate that pA47 is synthesized by Mc and epidermal cells as high molecular precursors. Using immunostaining techniques, we showed Mc to be involved in the formation of mesogleal non-collagenous (called "elastic" in classic morphological studies) fibres. The biochemical and morphological data suggest that Mc originate from the epidermis.     

Food and feeding of Aurelia aurita in Tokyo Bay with an analysis of stomach contents and a measurement of digestion times

In situ seasonal variations in stomach contents of Aurelia aurita (L.) in Tokyo Bay, Japan, were analyzed. Copepods, such as Oithona davisae Ferrari & Orsi were the predominant food items of A. aurita from June to November. The mean digestion time measured in incubation experiments was 0.95 h. Daily rations calculated using stomach content data and digestion times were 2.2–21.8 mg C ind^–1 corresponding to 0.58–5.56% of body carbon. The ingestion rate increased significantly with an increase in medusa size, although no significant relationship was found between medusa size and carbon specific daily ration. The zooplankton community in Tokyo Bay is characterized by the significant dominance of O. davisae and it is assumed that the prosperity of A. aurita is caused by the high abundance of the O. davisae population. It is suggested that a food chain comprised of microflagellates, cyclopoid copepods O. davisae, and A. aurita is the most significant one in Tokyo Bay and only a small portion of production is transferred to fish.     

Molecular characterization of aspartylglucosaminidase,a lysosomal hydrolase upregulated during strobilation in the moon jellyfish,Aurelia aurita

The life cycle of the moon jellyfish, Aurelia aurita, alternates between a benthic asexual polyp stage and a planktonic sexual medusa (jellyfish) stage. Transition from polyp to medusa is called strobilation. To investigate the molecular mechanisms of strobilation, we screened for genes that are upregulated during strobilation using the differential display method and we identified aspartylglucosaminidase (AGA), which encodes a lysosomal hydrolase. Similar to AGAs from other species, Aurelia AGA possessed an N-terminal signal peptide and potential N-glycosylation sites. The genomic region of Aurelia AGA was approximately 9.8 kb in length and contained 12 exons and 11 introns. Quantitative RT-PCR analysis revealed that AGA expression increased during strobilation, and was then decreased in medusae. To inhibit AGA function, we administered the lysosomal acidification inhibitors, chloroquine or bafilomycin A1, to animals during strobilation. Both inhibitors disturbed medusa morphogenesis at the oral end, suggesting involvement of lysosomal hydrolases in strobilation.     

5alpha, 8alpha-Epidioxysterol sulfate from a diatom Odontella aurita

A 5alpha,8alpha-epidioxysterol sulfate was isolated from the cultured diatom Odontella aurita (NIES 589), and its structure was elucidated by spectroscopic methods.     

Formation of Chaotic Patterns of the Gastrovascular System in the Ontogenesis of the Medusa Aurelia aurita

Only the first branching of the perradial and interradial channels of the gastrovascular system of Aurelia aurita (L.) (Scyphozoa: Ulmaridae) in the metaephyra stage is always uniform. Further branch formation is variable and random. New branches originate most often at the annular channel, but some of them grow from the radial channels. The early formation of irregular and chaotic patterns of the radial channel branching is caused by the asynchrony and topographic variability of the generation of new branches of different channels. During further morphogenesis, the spatial and temporal variability of branching increases, which leads to an increased state of chaos in the morphology of the branching channels of each individual. Lack of rigid determinancy of the gastrovascular system channel branching and plasticity of this system in the course of the entire ontogenesis provide possible evidence of its adaptive responses, for example, to the disruption of tetraradial symmetry or rearrangements after damage.     

Organization of the subumbrellar musculature in the ephyra,juvenile, and adult stages of Aurelia aurita Medusae

We used fluorescently labeled phalloidin to examine the subumbrellar musculature of the scyphozoan jellyfish Aurelia aurita in a developmental series from ephyra to adult medusa. In the ephyra, the swim musculature includes a disc‐like sheet of circular muscle, in addition to two radial bands of muscle in each of the eight ephyral arms. The radial muscle bands join with the circular muscle, and both circular and radial muscle act together during each swim contraction. As the ephyra grows into a juvenile medusa, arms tissue is resorbed as the bell tissue grows outward, so eventually, the ephyral arms disappear. During this process, the circular muscle disc also grows outward and the radial muscle bands of the arms also disappear. At this time, a marginal gap appears at the bell margin, which is devoid of circular muscle cells, but has a loose arrangement of radial muscle fibers. This marginal gap is preserved as the medusa grows, and contributes to the floppy nature of the bell margin. Radial distortions in the circular muscle layer involve muscle fibers that run in random directions, with a primarily radial orientation. These are believed to be remnants of the radial muscle of the ephyral arms, and the distortions decrease in number and extent as the medusa grows. Since the mechanics of swimming changes from drag‐based paddling in the ephyra to marginal rowing in the adult medusa, the development of the marginal gap and the presence of radial distortions should be considered in terms of this mechanical transition.     

Behavioural responses of zooplankton to the presence of predatory jellyfish

The major objective of this study was to investigate the behavioural responses of several zooplankton species to the presence of the scyphozoan jellyfish, Catostylus mosaicus. Specific aims included: identifying taxa that were not captured by C. mosaicus; investigating whether some of these taxa were able to detect and avoid water that had been exposed to C. mosaicus; and determining if positive phototaxis of crab megalopae was suppressed in the presence of C. mosaicus. Zooplankton not caught by C. mosaicus were identified by comparing the zooplankton present in the water column to those on the oral arms of the jellyfish. C. mosaicus mainly caught mollusc veligers and copepods, but did not catch crab megalopae, small prawns or post-flexion fish larvae. The hypothesis that these taxa were able to avoid swimming in water exposed to C. mosaicus was tested using water-choice experiments in a flume tank. A significant proportion (18-25%) of larval barramundi (Lates calcarifer) avoided swimming in the plume of water that had been exposed to C. mosaicus but mud crab (Scylla serrata) megalopae and juvenile prawns showed no response. The effect of C. mosaicus on the positive phototaxis of S. serrata megalopae was tested using 1 m tall glass towers. Megalopae were exposed to one of four treatments: filtered seawater (a control), an oral arm of C. mosaicus, an oral arm that had been sealed in plastic, and mucus from C. mosaicus. Megalopae migrated higher into the water column in the control than in treatments containing cues from the jellyfish. These findings suggest that blooms of jellyfish may induce behavioural changes in some zooplankton.     

Two-step purification and in vitro characterization of a hemolysin from the venom of jellyfish Cyanea nozakii Kishinouye

Hemolysin is one of the most hazardous components in the venom of Cyanea nozakii Kishinouye. Here we describe the purification and in vitro characterization of the hemolysin, which we named CnPH. The CnPH was isolated by anion-exchange and size-exclusion chromatography from the nematocyst venom. Two protein bands with molecular masses of 20 kDa, 60 kDa respectively were shown in the reducing SDS-PAGE analysis of the CnPH. And Approximately 5 μg/mL of the CnPH resulted in 50% hemolysis of the erythrocyte suspension. The hemolytic activity of the CnPH was both temperature and pH dependent. Moreover, it was significantly inhibited in the presence of divalent metal cations, including Cu²⁺, Mg²⁺, Mn²⁺, Zn²⁺ and Ca²⁺, but enhanced in the presence of EDTA. However, how CnPH performs its hemolytic activity is not yet clear, therefore the mechanism of the hemolytic activity of the CnPH is under research.