Changes in axonal transport in neurones of Asterias vulgaris and Asterias forbesei produced by colchicine and dimethyl sulfoxide.
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Complement-like activity in the sea star, Asterias forbesi.
The coelomic fluid (CF) of Asterias forbesi was examined for the presence of complement-like activity. Cell-free CF contains hemolytic activity for unsensitized rabbit erythrocytes (E) but not unsensitized or antibody-coated sheep E. Target E sensitivity is not dependent upon phylogenetic proximity of erythrocyte species to Asterias. The activity is optimally assayed at 25 degrees C, has a sigmoidal dose-response curve, and is heat-sensitive (56 degrees C). The activity is divalent cation dependent, although Ca++ and Mg++ are not required. Nonlytic CF does not contain an inhibitor; rather, combining nonlytic and lytic CFs results in an increase in hemolysis. Activity can be consumed by treatment with zymosan, pronase, trypsin, and PMSF. Enzymatic desialidation converts sheep E into activators of the hemolytic pathway. No phospholipase activity is present in CF. Biological function is demonstrated by bactericidal activity against Vibrio tubiashii by lytic CF. These results suggest that a primitive complement system is operating in Asterias.
Animals↗
Catalytic properties of the CMP-N-acetylneuraminic acid hydroxylase from the starfish Asterias rubens: comparison with the mammalian enzyme.
The biosynthesis of N-glycolylneuraminic acid (Neu5Gc) was investigated in cell-free extracts of the starfish Asterias rubens, which is one of the evolutionarily least-advanced species known to possess Neu5Gc-containing glycoconjugates. As in higher animals, Neu5Gc is synthesised in Asterias rubens by the action of a CMP-Neu5Ac hydroxylase. Enzyme activity was detected in all starfish tissues tested, the turnover being the greatest in the gonads. The enzyme from this tissue has a temperature optimum between 25 and 33 degrees C and a pH optimum between pH 6.0 and 6.4. This hydroxylase exhibits many characteristics in common with the mammalian enzyme. For example, the enzyme is extracted in a predominantly soluble form. Oxygen and a reduced pyridine nucleotide are necessary for activity, with NADH being the most effective cofactor. Furthermore, the activation of the hydroxylase by exogenously added iron salts and the potent inhibitory effects of several iron ligands point to the involvement of a non-haem iron cofactor. The enzyme has a high affinity for the substrate CMP-Neu5Ac, the apparent Km being 18 microM. In contrast to the mammalian enzyme, the hydroxylase from Asterias rubens is not inhibited by increased ionic strength and cannot be activated by non-ionic detergents. Moreover, the CMP-Neu5Ac turnover increased linearily with increasing protein concentration. In accordance with other enzymes in starfish, seasonal changes in the CMP-Neu5Ac hydroxylase activity were also observed.
Animals↗
Immunocytochemical mapping of the novel echinoderm neuropeptide SALMFamide 1 (S1) in the starfish Asterias rubens.
The recent isolation and characterization of the SALMFamide neuropeptides S1 and S2 from the starfish Asterias rubens has initiated a series of studies on their distribution. Specific antisera have been raised against S1 and used in light-microscopical immunocytochemistry. The results of this study reveal for the first time a possible hyponeural innervation of the visceral musculature of the gut and the widespread neuronal distribution of S1, (i) in axons and cell bodies of both ectoneural and hyponeural regions of the radial nerve cord and circumoral nerve ring, (ii) in the nerve ring and nerve plexus of the tube feet, (iii) in the apical muscle, (iv) in skin, and (v) extensively throughout the digestive system. These discoveries are of particular interest in terms of the possible functional roles for S1 in Asterias rubens.
Amino Acid Sequence↗
Protein phosphorylation during 1-methyladenine-induced maturation of Asterias oocytes.
Maturation was induced in Asterias oocytes with 1-methyladenine (1-MA) at a final concentration of 2 microM. At 5, 10, and 30 min of treatment, oocytes were homogenized and the cytosolic fraction was prepared. The cytosol was incubated with [gamma-32P]ATP and [gamma-32P]GTP. The phosphorylated proteins were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and the radioactivity in the gels was determined by autoradiography. The cytosol prepared from 1-MA-treated oocytes incubated with [gamma-32P]ATP showed a marked increase in the radiolabeling of proteins with estimated molecular weights of 70,000 and 62,000 Da. With [gamma-32P]GTP a 56,000-Da protein showed increased radiolabeling. The present finding suggests that an early biochemical event of 1-MA-induced oocyte maturation in Asterias is the stimulation of phosphorylation of specific proteins.
Adenine↗
Enzymatic properties of a new type of lysozyme isolated from Asterias rubens : comparison with the Nephthys hombergii (annelid) and hen lysozymes.
The enzymatic properties of Asterias rubens and Nephthys hombergii lysozymes were compared with those of hen egg-white lysozyme. The results allowed to conclude that the Asterias rubens lysozyme was representative of a new lysozyme type. The Nephthys hombergii lysozyme however could be classified among the hen type enzymes with a tendency to exhibit under certain conditions some properties attributed to the goose type lysozymes.
Acetylglucosamine↗
The lysozyme of the starfish Asterias rubens. A paradygmatic type i lysozyme.
On the basis of a partial N-terminal sequence, Jollès and Jollès previously proposed that the lysozyme from the starfish Asterias rubens represents a new form of lysozyme, called type i (invertebrate) lysozyme. Indeed, it differed from both the types c (chicken) and g (goose) known in other animals, as well as from plant and phage lysozymes. Recently, several proteins belonging to the same family have been isolated from protostomes. Here we report the complete mature protein sequence and cDNA sequence of the lysozyme from Asterias. These sequences vindicate the previously proposed homology between the starfish, a deuterostome, and protostome lysozymes. In addition, we present a structural analysis that allows us to postulate upon the function of several conserved residues.
Amino Acid Sequence↗
Tissue distribution of the SALMFamide neuropeptides S1 and S2 in the starfish Asterias rubens using novel monoclonal and polyclonal antibodies. I. Nervous and locomotory systems.
The recent isolation and characterization of the SALMFamide neuropeptides (S1 and S2) from the starfish Asterias rubens has initiated a series of single- and double-labelling immunocytochemical studies to ascertain their tissue distribution and cellular localization. Specific novel monoclonal and polyclonal antisera have been raised against these neuropeptides and used in optical immunocytochemistry (ICC). The results of the present study reveal, for the first time, the widespread neuronal distribution of S2 localized to axons and perikarya of the radial nerve cord and tube foot ectoneural nerve plexus. Double labelling revealed a predominantly separate localization for S1 and S2 immunoreactivity. The potential functional roles of S1 and S2 in the radial nerve cord (RNC) and tube feet of Asterias rubens are discussed.
Amino Acid Sequence↗
Biogeography of Asterias: North Atlantic climate change and speciation.
Fossil evidence suggests that the seastar genus Asterias arrived in the North Atlantic during the trans-Arctic interchange around 3.5 Ma. Previous genetic and morphological studies of the two species found in the Atlantic today suggested two possible scenarios for the speciation of A. rubens and A. forbesi. Through phylogenetic and population genetic analysis of data from a portion of the cytochrome oxidase I mitochondrial gene and a fragment of the ribosomal internal transcribed spacer region, I show that the formation of the Labrador Current 3.0 Ma was probably responsible for the initial vicariance of North Atlantic Asterias populations. Subsequent adaptive evolution in A. forbesi was then possible in isolation from the European species A. rubens. The contact zone between these two species formed recently, possibly due to a Holocene founding event of A. rubens in New England and the Canadian Maritimes.
Animals↗
Gamete compatibility and sperm competition affect paternity and hybridization between sympatric Asterias sea stars.
Gamete interactions may strongly influence speciation and hybridization in sympatric broadcast-spawning marine invertebrates. We examined the role of gamete compatibility in species integrity using cross-fertilization studies between sympatric Asterias sea stars from a secondary contact zone in the northwest Atlantic. In crosses between single males and single females, gametes of both species were compatible and produced viable, fertile hybrid offspring, but with considerable variation in the receptivity of eggs to heterospecific sperm. Differential compatibility of heterospecific gametes was detected in sperm competition studies in which we used a nuclear DNA marker to assign paternity to larval offspring. Several families showed conspecific sperm precedence in A. forbesi eggs, and one family showed competitive superiority of A. forbesi sperm fertilizing A. rubens eggs. Gametic interactions are an important component of prezygotic reproductive isolation in sympatric Asterias. The interaction between gametes of these closely related sea stars is consistent with the function of gamete recognition systems that are known to mediate fertilization success and speciation in other marine invertebrates.
Animals↗
Clotting enzyme activity derived from the coelomocytes of the sea star Asterias forbesi.
Clotting enzyme activity was detected in lysates prepared from the coelomocytes of Asterias forbesi following incubation with endotoxin-activated Limulus amebocyte lysate. This activity was not detected in the cell-free coelomic fluid. The enzymatic activity from the sea star lysate hydrolyzed the synthetic substrate S2222 but not S2238 or S2251, and polymerized partially purified Limulus clottable protein. The clotting enzyme activity was not detected following treatment of the sea star cell lysate with endotoxin or with the clotting enzyme from Limulus lysate. The enzymatic activity, generated in sea star cell lysate by the activated Limulus lysate, was inhibited by the addition of benzamidine; and its effect was suppressed with rabbit anti-sera directed against Asterias whole cell lysate, but not with anti-sera directed against the previously reported Sea Star Factor.
Animals↗
Alteration and recovery of appetitive behaviour following nerve section in the starfish Asterias rubens.
The starfish Asterias rubens is an invertebrate deuterostome whose nervous system shows remarkable regenerative properties. To understand when full functionality of a damaged part of the nervous system recovers, and to follow nerve regeneration in detail, we carried out behavioural experiments with 29 starfishes that had the nerve in one of the arms sectioned in a mid-arm position. Loss and recovery of normal behaviour was followed by video analysis of animal performance in an appetitive behavioural test. When compared to 13 control (unoperated) animals, the appetitive response of freshly sectioned animals is normal initially, progressively deteriorates up to 40 days after the lesion, and then gradually improves until 60 days, when recovery is complete. This is true only when one of the leading arms in the appetitive test is a sectioned arm; turning the starfish so that both the leading arms facing the prey are unlesioned, results in normal behaviour even at 40 days after the cut. Thus, regeneration is a multi-step process whose time course coincides with anatomical regeneration. At intermediate times the animals have coordination problems in an appetitive behaviour test and these give some insights into how arms may inter-communicate to organize concerted movements.
Animals↗
Analysis of monosaccharides, fatty constituents and rare O-acetylated sialic acids from gonads of the starfish Asterias rubens.
A previous study (Bergwerff et al., Biochimie 74 (1992) 25-37) reported that sialic acids present in Asterias rubens gonads were essentially composed of 8-methyl-N-glycolylneuraminic acid (Neu5Gc8Me), a large part of it being acetylated in position 9. Using GC/MS of heptafluorobutyrate derivatives (Zanetta et al., Glycobiology 11 (2001) 663-676) on the chloroform/methanol soluble and insoluble fractions, we showed that most sialic acids were found in the latter and demonstrated that all sialic acids were derived from N-glycolylneuraminic acid, most of them being 8-methylated, but that the majority were also acetylated in position 4 or 7 (or both positions). GC/MS analyses of the constituents liberated using acid-catalysed methanolysis verified that major glycoprotein-bound glycans were N-linked and of the gluco-oligomannosidic type. Major fatty acids were poly-unsaturated (especially C20:4) and long-chain bases were C22:1 phytosphingosine and C22:2 6-hydroxysphingenine. Major monosaccharides found in the chloroform/methanol extract (quinovose and fucose) were derived from steroidal saponins.
Animals↗
Bioaccumulation and effects of PCBs and heavy metals in sea stars (Asterias rubens, L.) from the North Sea: a small scale perspective.
Sea stars (Asterias rubens L.) were collected in different stations distributed in the Southern Bight of the North Sea. Concentrations of four heavy metals and six PCB congeners were measured in two body compartments (body wall and pyloric caeca). In order to assess the potential harm of these contaminants, two biochemical parameters were measured in sea stars, viz. reactive oxygen species (ROS) production by amoebocytes and cytochrome P450 immunopositive protein (CYP1A IPP) induction in pyloric caeca. Sea stars from stations located in the plume of the Scheldt river showed the highest contamination levels. Other stations, similarly located, displayed lower levels. No simple relationship could be established between ROS production by sea star amoebocytes and contaminant levels measured in sea star tissues. CYP1A IPP induction displayed more contrasted responses, and highly significant regressions were found between PCB concentrations measured in pyloric caeca and CYP1A IPP. Both biological parameters were found to vary significantly over the study area. On the whole, data indicated that contamination levels and subsequent effects in sea stars were comparable to those described in previous large-scale studies, but that working at a smaller scale highlighted the existence of patterns of contamination which can blur general trends due to major contamination sources like contaminated rivers.
Animals↗
A new triterpene glycoside from Asterias rollentoni.
A new triterpene glycoside, rollentoside A, has been isolated from Asterias rollentoni Bell and identified as 3beta-O-{3-O-methyl-beta-D-xylopyranosyl-(1-->3)-O-beta-D-glucopyranosyl-(1-->4)-O-beta-D-quinopyranosyl-(1-->2)-O-beta-D-xylopyranosyl}-16-beta-acetoxy-23S-acetoxy-holost-7-ene (1), together with a new natural product, rollentoside B (2). The structures of compounds1 and 2 were elucidated by extensive 1D and 2D NMR investigation (1H-1H COSY, TOCSY, HSQC, HMBC, NOESY).
Animals↗
A chemical and histochemical study of the lipides of the pyloric cecum of the starfish, Asterias forbesi.
The lipides of the diverticula of Asterias forbesi have been studied by histochemical and biochemical means. Correlations between results obtained by histochemical examination of sections, and chemical analysis of isolated lipide have been made, particularly with respect to phosphatides, steroids, and aldehyde lipides. The results of the histochemical study were in good agreement with the chemical data as to the nature of the phosphatide fraction, the presence of acetone-soluble aldehyde lipides, and the composition of the free droplet fat. Homogenized diverticula were differentially centrifuged in order to establish the distribution of types of lipides in the various cellular components. In addition, data have been presented which demonstrate a direct correlation between the titer of alpha-glycerol ethers and that of acetone-soluble lipide acetals in the unsaponifiable fraction.
Animals↗
Computer-assisted evaluation of isoelectric focusing patterns in electrophoretic gels: identification of smoothhounds (Mustelus mustelus, Mustelus asterias) and comparison with lower value shark species.
In this work, the most commercially important Selachian species, smoothhound (Mustelus mustelus) and starry smoothhound (Mustelus asterias), have been identified by polyacrylamide isoelectric focusing (IEF-PAGE), along with several shark species of minor commercial value. In Italy, these two smoothhound species are commonly subjected to fraudulent substitution with lesser valued sharks. After the electrophoretic runs, the band patterns of the two Mustelus species were compared with those of dogfish (Scyliorhinus canicula), spurdog (Squalus acanthias), blue shark (Prionace glauca) and black-mouthed dogfish (Galeus melanostomus), both visually and with gel analysis software. The actual isoelectric points were then submitted to cluster analysis to differentiate the single species, despite the possible occurrence of electrophoretic variations or protein polymorphism. Every shark showed species-specific band patterns and could therefore be well differentiated, as confirmed by statistical analysis.
Acrylic Resins↗
Purification of an antibody-like protein from the sea star Asterias rubens (L.).
Cells from sea star (Asterias rubens) axial organs stimulated with trinitrophenyl (TNP) or fluoresceinyl-haptened polyacrylamide beads and subsequently stimulated in vitro with the same antigen produced and released a specific antibody-like protein which induced lysis of haptened sheep erythrocytes in the presence of serum complement. The anti-TNP antibody-like protein isolated by ammonium sulfate precipitation, gel filtration and affinity chromatography exhibited a single precipiting peak after crossed immunoelectrophoresis against rabbit antiserum to partially purified culture supernatant. The anti-TNP antibody-like protein gave a specific affinity precipitate in crossed affino-electrophoresis using a p-nitrobenzoyl-substituted gel. The analysis by sodium dodecyl sulfate-polyacrylamide gel electrophoresis under both reducing and nonreducing conditions evidenced a unique 30-kDa polypeptide chain. According to gel filtration experiments, the molecular weight of the major component isolated by affinity chromatography was about four times higher. Therefore, the antibody-like molecule could be a tetrameric protein devoid of any disulfide bond.
Animals↗