Who knows not where an anemone does wear his sting? Could polypeptides released from the columnar vesicles of Bunodosoma cangicum induce apoptosis in the ZF-L cell line?

We provide ultrastructural and cytological evidence that the tentacles of the sea anemone Bunodosoma cangicum does not contain cytotoxic venom. However, we show that the stimulated secretion of an apparent mixture of biomolecules containing polypeptides from the columnar vesicles of Bunodosoma cangicum is apparently a potent inducer of apoptosis in the zebrafish cell line, ZF-L. Microscopic fluorescence, cell morphology and flow cytometric assays confirm the apoptotic activity. Crude vesicle venom was partially purified by size exclusion chromatography. PAGE analysis shows that this venom contains low weight polypeptides but no measurable protein. The apoptotic activity is heat labile, and the observed peptides concurrent with this activity have a molecular weight of approximately 2000 Da. This manuscript is the first report of biologically active molecules and peptides associated with columnar vesicles of anemones, and the first to confirm that the tentacles of B. cangicum do not contain cytotoxic venom, and express spirocytes exclusively.

Quantitative study of proteomic alterations in a Zebrafish (danio rerio) cell line infected with the Singapore Grouper Iridovirus (SGIV).

We demonstrate, for the first time, that Singapore Grouper Iridovirus (SGIV) can successfully infect a Zebrafish cell line. Combined with the recent availability of the complete zebrafish (danio rerio) genome, this provides an opportunity to investigate virus-host interactions at the molecular level. Using iTRAQ labeling and two-dimensional LC/MS/MS quantitative proteomics, 157 zebrafish proteins exhibiting significant alterations in expression levels following SGIV infection were identified. Gene ontology analysis revealed that SGIV controls a wide aspect of zebrafish host machinery to ensure replication and propagation. In order to probe the mechanism underlying SGIV infection in Zebrafish cells, we used an anti-sense morpholino to knockdown orf86r, an immediate early viral gene that encodes the SGIV protein ORF86R. The expression profile of certain host proteins involved in replication was altered upon knockdown. In particular, expression of CNOT, a non-enzymatic subunit of the CCR4-NOT transcription complex was markedly affected. Taken together, these findings provide a new insight on the function of the essential viral protein ORF86R. Our results show that Singapore Grouper Iridovirus infection of a Zebrafish cell line is a useful new tool to study virus-host interactions.

Zebrafish liver (ZFL) cells are able to mount an anti-viral response after stimulation with Poly (I:C).

The zebrafish (Danio rerio) is a widely used model species for biomedical research and is also starting to be a model for aquaculture research. The ZFL cell line, established from zebrafish liver, has been mostly used in toxicological and ecotoxicological studies. However, no studies have previously characterised this cell line in regard to its immunological response. The aim of this work was to study the gene expression response of the ZFL cell line after incubation with different prototypical immune stimuli, such as lipopolysaccharide (LPS), peptidoglycan (PGN), zymosan, and with a special focus on the dsRNA Poly (I:C). Using PCR, microarrays, and confocal microscopy we have explored the response of the ZFL cells against Poly (I:C). This study shows that the ZFL is able to uptake very efficiently the Poly (I:C) and mount a strong anti-viral response. We can conclude that ZFL could be used not only in toxicological studies, but also in studying anti-viral responses in zebrafish.