Purification, characterization and cDNA cloning of a lectin from the brittle star Ophioplocus japonicus.

Lectins are carbohydrate-binding proteins that possess specific sugar-binding properties and are involved in various biological activities in different organisms. In this study, purification, characterization, and cDNA cloning of a brittle star lectin, designated as Ophioplocus japonicus agglutinin (OJA), were conducted. OJA was isolated from the brittle star O. japonicus by affinity chromatography on a Sephadex G-25 column, followed by ion-exchange chromatography on a Resource Q column. This lectin yielded distinct bands at approximately 176 or 17 kDa on sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) under non-reducing or reducing conditions, respectively. It also exhibited Ca2+-dependent hemagglutination activity, which, however, was not affected by other metal cations, such as Ba2+, Co2+, Cu2+, Zn2+, Fe2+, Mg2+, and Mn2+. The OJA activity was strongly inhibited by glucose and xylose among the monosaccharides tested, and by bovine thyroglobulin among the glycoproteins tested. Cloning of the OJA cDNA revealed that its primary structure contained the C-type lectin domain (CTLD). The results of this study showed that OJA is an echinoderm-derived glucose/xylose-specific lectin that belongs to the C-type lectin superfamily.

Molecular identification of larval bucephalids, Prosorhynchoides ozakii and Parabucephalopsis parasiluri , infecting the golden mussel, Limnoperna fortunei , by PCR-RFLP.

A polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) technique was developed for the molecular identification of 2 introduced bucephalid trematodes, Prosorhynchoides ozakii and Parabucephalopsis parasiluri . The method was applied for sporocysts and cercariae obtained from the golden mussel Limnoperna fortunei collected in the Uji River, Japan. The PCR-RFLP method showed that L. fortunei is the intermediate host of both trematode species. The present study thus recognizes the risk of L. fortunei , an invasive molluscan species, as a potential host for pathogenic trematodes.

Liolope copulans (Trematoda: Digenea: Liolopidae) parasitic in Andrias japonicus (Amphibia: Caudata: Cryptobranchidae) in Japan: Life cycle and systematic position inferred from morphological and molecular evidence.

The life cycle of Liolope copulans Cohn, 1902 (Trematoda: Digenea: Liolopidae), an intestinal parasite of the Japanese giant salamander Andrias japonicus (Temminck) (Amphibia: Caudata: Cryptobranchidae), was studied in the field and laboratory in Japan. This is the first description of mother sporocyst, daughter sporocyst and cercariae of a liolopid species. Non-oculate longifurcate pharyngeate cercariae were formed in lanceolate-cylindrical daughter sporocysts in Semisulcospira libertina (Gould) (Gastropoda: Sorbeoconcha: Pleuroceridae). They successfully developed to encapsulated metacercariae in cyprinid fishes, Nipponocypris sieboldii (Temminck and Schlegel) and Rhynchocypris lagowskii (Dybowski), by experimental infection. Cercariae had a V-shaped excretory vesicle with two looped arms, as in metacercariae and adults. Developmental stages from mother sporocyst to adult are described and illustrated. DNA sequencing was conducted for 28S and 18S rDNA of mother and daughter sporocysts, cercariae, and an adult. The result of molecular phylogenetic analysis suggests that L. copulans may be one of the basal taxa of the order Diplostomida Olson, Cribb, Tkach, Bray, and Littlewood, 2003, but its systematic position is still unclear because of the topological inconsistence between the 28S and 18S trees. Therefore, we tentatively place the family Liolopidae in the superfamily Diplostomoidea, mainly based on the morphology of sporocysts and cercariae.

Possible role of a taurine transporter in the deep-sea mussel Bathymodiolus septemdierum in adaptation to hydrothermal vents.

Various invertebrates inhabiting hydrothermal vents possess sulfur-oxidizing bacteria in their tissues; however, the mechanisms by which toxic sulfides are delivered to these endosymbionts remain unknown. Recently, detoxification of sulfides using thiotaurine, a sulfur-containing amino acid, has been suggested. In this study, we propose the involvement of a taurine transporter in sulfide detoxification in the deep-sea mussel Bathymodiolus septemdierum by demonstrating: (i) the abundance of its mRNA in the gill; (ii) its activity under a wide range of salinities; (iii) its low Michaelis constant value in taurine transportation; and (iv) its affinity for thiotaurine and the thiotaurine precursor, hypotaurine.

Expression and functional analysis of mussel taurine transporter, as a key molecule in cellular osmoconforming.

Most aquatic invertebrates adapt to environmental osmotic changes primarily by the cellular osmoconforming process, in which osmolytes accumulated in their cells play an essential role. Taurine is one of the most widely utilized osmolytes and the most abundant in many molluscs. Here, we report the structure, function and expression of the taurine transporter in the Mediterranean blue mussel (muTAUT), as a key molecule in the cellular osmoconforming process. Deduced amino acid sequence identity among muTAUT and vertebrate taurine transporters is lower (47-51%) than that among vertebrate taurine transporters (>78%). muTAUT has a lower affinity and specificity for taurine and a requirement for higher NaCl concentration than vertebrate taurine transporters. This seems to reflect the internal environment of the mussel; higher NaCl and taurine concentrations. In addition to the hyperosmotic induction that has been reported for cloned taurine transporters, the increase in muTAUT mRNA was unexpectedly observed under hypoosmolality, which was depressed by the addition of taurine to ambient seawater. In view of the decrease in taurine content in mussel tissue under conditions of hypoosmolality reported previously, our results lead to the conclusion that muTAUT does not respond directly to hypoosmolality, but to the consequent decrease in taurine content. By immunohistochemistry, intensive expression of muTAUT was observed in the gill and epithelium of the mantle, which were directly exposed to intensive osmotic changes of ambient seawater.

Scallop DMT functions as a Ca2+ transporter.

We identified a DMT (divalent metal transporter) homologous protein that functions as a Ca(2+) transporter. Scallop DMT cDNA encodes a 539-amino-acid protein with 12 putative membrane-spanning domains and has a consensus transport motif in the fourth extracellular loop. Since its mRNA is significantly expressed in the gill and intestine, it is assumed that scallop DMT transports Ca(2+) from seawater by the gill and from food by the intestine. Scallop DMT lacks the iron-responsive element commonly found in iron-regulatory proteins, suggesting that it is free of the post-transcriptional regulation from intracellular Fe(2+) concentration. Scallop DMT distinctly functions as a Ca(2+) transporter unlike other DMTs, however, it also transports Fe(2+) and Cd(2+) similar to them.