RNA Aptamers Inhibit the Growth of the Fish Pathogen Viral Hemorrhagic Septicemia Virus (VHSV)

Viral hemorrhagic septicemia virus (VHSV) is a serious disease impacting wild and cultured fish worldwide. Hence, an effective therapeutic method against VHSV infection needs to be developed. Aptamer technology is a new and promising method for diagnostics and therapeutics. It revolves around the use of an aptamer molecule, an artificial ligand (nucleic acid or protein), which has the capacity to recognize target molecules with high affinity and specificity. Here, we aimed at selecting RNA aptamers that can specifically bind to and inhibit the growth of a strain of fish VHSV both in vitro and in vivo. Three VHSV-specific RNA aptamers (F1, F2, and C6) were selected from a pool of artificially and randomly produced oligonucleotides using systematic evolution of ligands by exponential enrichment. The three RNA aptamers showed obvious binding to VHSV in an electrophoretic mobility shift assay but not to other tested viruses. The RNA aptamers were tested for their ability to inhibit VHSV in vitro using hirame natural embryo (HINAE) cells. Cytopathic effect and plaque assays showed that all aptamers inhibited the growth of VHSV in HINAE cells. In vivo tests using RNA aptamers produced by Rhodovulum sulfidophilum showed that extracellular RNA aptamers inhibited VHSV infection in Japanese flounder. These results suggest that the RNA aptamers are a useful tool for protection against VHSV infection in Japanese flounder.     

Permissive Effect of Insulin-like Growth Factor I (IGF-I) on Gonadotropin Releasing-hormone Action on In Vitro Growth Hormone Release,in Rainbow Trout (Oncorhynchus mykiss)

The aim of the work was to study the influence of insulin-like growth factor I (IGF-I) on GnRH-induced GH release by cultured pituitary cells of normally growing rainbow trout (Oncorhynchus mykiss), collected at different stages of gametogenesis. When pituitary cells were pre-incubated with human IGF-I (10⁻⁸ M) for 48 hours they became responsive to sGnRH (10⁻⁸ to 10⁻⁶ M) in the subsequent 24-hour incubation period, depending on the sexual stage, while not IGF-I pre-incubated cells were always non-responsive to GnRH. The permissive effect of IGF-I was detected in immature fish or those at the beginning of the gametogenesis, but not in mature fish. IGF-I inhibition of GH release during the preincubation period varies also with the sexual stage and is greater in immature than in mature fish. The permissive effect of IGF-I seems specific to somatotropes since IGF-I does not modify GnRH action on GtH₂ release. This work suggests that GnRH action on GH release can vary for a particular fish species depending on the physiological status.