Suppression substractive hybridisation (SSH) and real time PCR reveal differential gene expression in the Pacific cupped oyster, Crassostrea gigas, challenged with Ostreid herpesvirus 1

Renault, Tristan; Faury, Nicole; Barbosa-Solomieu, Valérie; Moreau, Kévin

doi:10.1016/j.dci.2011.02.004

Cited by 144 publications

(140 citation statements)

References 56 publications

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“…The concentration of cells was adjusted to 3 ϫ 10 6 cells/ml in FSW, and 1 ml of the cell suspension was dispensed in each well of a 24-well plate. The plates were incubated at 15°C for 1 h for settlement and further infected following the procedure previously described by Renault et al (22). All experiments were performed at 15°C.…”

Section: Methodsmentioning

confidence: 99%

“…The OsHV-1 Var was kindly provided by Tristan Renault (IFREMER, France) as a viral suspension obtained from infected oyster hemocytes (22). An HSV-1 clinical isolate (Ͼ99% homology to the isolate SK087 US4-6 genes; GenBank accession number AY240815.1) and an HSV-2 clinical isolate (Ͼ99% homology to the isolate 99-62039 US4 gene; GenBank accession number HM011430.1) were provided by the Virology Service of Hospital Universitario Central de Asturias (Oviedo, Spain), propagated in Vero cells (European Collection of Authenticated Cell Cultures [ECACC] no.…”

Section: Methodsmentioning

confidence: 99%

See 1 more Smart Citation

Antiviral Activity of Myticin C Peptide from Mussel: an Ancient Defense against Herpesviruses

Novoa

Romero

Álvarez

et al. 2016

J Virol

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Little is known about the antiviral response in mollusks. As in other invertebrates, the interferon signaling pathways have not been identified, and in fact, there is a debate about whether invertebrates possess antiviral immunity similar to that of vertebrates. In marine bivalves, due to their filtering activity, interaction with putative pathogens, including viruses, is very high, suggesting that they should have mechanisms to address these infections. In this study, we confirmed that constitutively expressed molecules in naive mussels confer resistance in oysters to ostreid herpesvirus 1 (OsHV-1) when oyster hemocytes are incubated with mussel hemolymph. Using a proteomic approach, myticin C peptides were identified in both mussel hemolymph and hemocytes. Myticins, antimicrobial peptides that have been previously characterized, were constitutively expressed in a fraction of mussel hemocytes and showed antiviral activity against OsHV-1, suggesting that these molecules could be responsible for the antiviral activity of mussel hemolymph. For the first time, a molecule from a bivalve has shown antiviral activity against a virus affecting mollusks. Moreover, myticin C peptides showed antiviral activity against human herpes simplex viruses 1 (HSV-1) and 2 (HSV-2). In summary, our work sheds light on the invertebrate antiviral immune response with the identification of a molecule with potential biotechnological applications. IMPORTANCESeveral bioactive molecules that have potential pharmaceutical or industrial applications have been identified and isolated from marine invertebrates. Myticin C, an antimicrobial peptide from the Mediterranean mussel (Mytilus galloprovincialis) that was identified by proteomic techniques in both mussel hemolymph and hemocytes, showed potential as an antiviral agent against ostreid herpesvirus 1 (OsHV-1), which represents a major threat to the oyster-farming sector. Both hemolymph from mussels and a myticin C peptide inhibited OsHV-1 replication in oyster hemocytes. Additionally, a modified peptide derived from myticin C or the nanoencapsulated normal peptide also showed antiviral activity against the human herpesviruses HSV-1 and HSV-2. Therefore, myticin C is an example of the biotechnological and therapeutic potential of mollusks.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Antiviral Activity of Myticin C Peptide from Mussel: an Ancient Defense against Herpesviruses

Novoa

Romero

Álvarez

et al. 2016

J Virol

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“…Numerous defence genes could also provide putative markers for selection (Gueguen et al 2003;Gonzales 2007;Gueguen et al 2009;Mateo et al 2010;Duperthuy et al 2011;Renault et al 2011). However, considering the proposed conceptual model, if their expression is limited by ROS production and related energy deficiency, it is probably better to prioritise the selection of genes preventing ROS production (strategy 1), or ROS detoxification genes (strategy 2) because these approaches will also control the expression of defence mechanisms.…”

Section: Is the Increased Mortality Observed In C Gigas Since 2008 Dmentioning

confidence: 99%

“…(2) different pathogens, such as bacteria (Vibrio splendidus, V. aestuarianus) and viruses: ostreid herpesvirus 1 (OsHV-1, and microvar OsHV-1 Renault et al 2011), infect the oysters; (3) very significant genetically-based resistance ("R" oysters) or susceptibility ("S" oysters) to summer mortality is possible, as revealed by divergent selection experiments. At the end of this program, the nature of the genetic mechanism that could form the basis of such a complex interaction and non-specific resistance remained a major question.…”

Section: Introductionmentioning

confidence: 99%

Review and perspectives of physiological mechanisms underlying genetically-based resistance of the Pacific oysterCrassostrea gigasto summer mortality

Samain

2011

Aquat. Living Resour.

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-French oyster farming has been subject to severe mortalities during the summer months. Results from the research program "Morest", which ran from 2000 to 2006 and examined the possible causes of these mortalities, led to the construction of a model to explain the interaction between environmental factors, oyster physiology and different opportunistic pathogens underlying oyster summer mortality. Temperature, food, reproduction and stress were the main factors required for oyster mortality. Genetically-based resistance ("R" oysters) or susceptibility ("S" oysters) to summer mortality was revealed by divergent selection. Building on these results, a literature search was made in 2007 on the molecular origin of genetic resistance to such a complex mortality risk. The objectives were to lay a foundation for the preparation and orientation of future research directions and to improve understanding of the underlying physiological mechanism leading to summer mortality. Three years later, the resulting conceptual analysis reported here was presented as an introductory lecture to Physiomar 2010, a conference where many new results contributing to this research field were also reported. The literature review highlighted two major review articles: the first dealing with nutrition and reproduction (Schneider 2004), the second with reproduction, temperature, oxidative stress and mortality (Heineinger 2002). The effect of nutrition level on energy orientation to growth or reproduction is controlled by endocrine factors. Among these, neuropeptide Y (NPY), ghrelin and leptin neuropeptides appeared to be potential candidates involved in germ-soma orientation in relation to trophic conditions. Depending on reproductive effort and temperature, a metabolic stress resulting from the germ-soma conflict can appear, characterized by mitochondrial reactive oxygen species (ROS) production. Such an excess of ROS induces perturbations in mitochondrial activity leading to cell death. Many organisms, such as annual plants or the Pacific salmon, do not survive their first reproduction. In contrast, others increase stress resistance by selection of antioxidant processes (superoxide dismutase SOD, catalase, etc.) through evolution, and survive first reproduction. A similar difference was observed in the comparison made between R and S oysters, which differed in ROS production, SOD and catalase levels. Such factors controlling reproduction and ROS detoxification processes could therefore provide new markers for selection of oysters with better resistance to non-specific pathogens, complementing other classic selection approaches against specific pathogens or for improved immunity. This antioxidant defence mechanism is found in many organisms including vertebrates and in some invertebrates, including oysters. Its role needs to be considered in pathology events involving other aquaculture species and it may also contribute to explaining the increase in marine pathologies under anthropogenic environmental changes.

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“…Suppression subtractive hybridization (SSH) combines normalization and subtraction in a single procedure, thus allowing the isolation of differentially expressed cDNA that is overexpressed in one population but not in another. In recent years, SSH has been successfully applied as an efficient method to identify differentially expressed genes in oyster (Renault et al, 2011), shrimp (He et al, 2004;Pan et al, 2005;Zhao et al, 2007;James et al, 2010), clam (Prado-Alvarez et al, 2009, and fish (Cui et al, 2011;Li et al, 2011b). However, these studies all focused on the identification of immune-related genes with the challenge of specific pathogens.…”

Section: Introductionmentioning

confidence: 99%

Identification and assessment of differentially expressed genes involved in growth regulation in Apostichopus japonicus

Zhu

Su³

et al. 2013

Genet. Mol. Res.

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ABSTRACT. Rapid and efficient growth is a major consideration and challenge for global mariculture. The differential growth rate of the sea cucumber, Apostichopus japonicus, has significantly hampered the total production of the industry. In the present study, forward and reverse suppression subtractive hybridization libraries were constructed and sequenced from a fast-growth group and a slow-growth group of the sea cucumber. A total of 142 differentially expressed sequence tags (ESTs) with insertions longer than 150 bp were identified and further analyzed. Fifty-seven of these ESTs (approximately 40%) were functionally annotated for cell structure, energy metabolism, immunity response, and growth factor categories. Six candidate genes, arginine kinase, cytochrome c oxidase subunit I, HSP70, β-actin, ferritin, and the ADP-ribosylation factor, were further validated by quantitative PCR. Significant differences were found between the fast-and slow-growth groups (P < 0.05) for the expression levels of arginine kinase, cytochrome 3029 ©FUNPEC-RP www.funpecrp.com.br Genetics and Molecular Research 12 (3): 3028-3037 (2013) Differentially expressed genes in A. japonicus growth c oxidase, HSP70, the ADP-ribosylation factor, and β-actin. However, no significant difference was observed for ferritin. Our results provide promising candidate gene markers for practical size screening, and also further promote marker-assisted selective breeding of this species.

show abstract

Suppression substractive hybridisation (SSH) and real time PCR reveal differential gene expression in the Pacific cupped oyster, Crassostrea gigas, challenged with Ostreid herpesvirus 1

Cited by 144 publications

References 56 publications

Antiviral Activity of Myticin C Peptide from Mussel: an Ancient Defense against Herpesviruses

Antiviral Activity of Myticin C Peptide from Mussel: an Ancient Defense against Herpesviruses

Review and perspectives of physiological mechanisms underlying genetically-based resistance of the Pacific oysterCrassostrea gigasto summer mortality

Identification and assessment of differentially expressed genes involved in growth regulation in Apostichopus japonicus

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