FN Archimer Export Format
PT J
TI Haemocytes collected from experimentally infected Pacific oysters, Crassostrea gigas: Detection of ostreid herpesvirus 1 DNA, RNA, and proteins in relation with inhibition of apoptosis
BT 
AF MARTENOT, Claire
   GERVAIS, Ophelie
   CHOLLET, Bruno
   HOUSSIN, Maryline
   RENAULT, Tristan
AS 1:1;2:1;3:1;4:2;5:3;
FF 1:PDG-RBE-SG2M-LGPMM;2:PDG-RBE-SG2M-LGPMM;3:PDG-RBE-SG2M-LGPMM;4:;5:PDG-RBE;
C1 IFREMER, Inst Francais Rech Exploitat Mer, Lab Genet & Pathol Mollusques Marins, La Tremblade, France.
   LABEO Frank Duncombe, Caen, France.
   IFREMER, Dept Ressources Biol & Environm, Nantes, France.
C2 IFREMER, FRANCE
   LABEO FRANK DUNCOMBE, FRANCE
   IFREMER, FRANCE
SI LA TREMBLADE
   NANTES
SE PDG-RBE-SG2M-LGPMM
   PDG-RBE
IN WOS Ifremer jusqu'en 2018
   DOAJ
   copubli-france
IF 2.766
TC 27
UR https://archimer.ifremer.fr/doc/00386/49715/50244.pdf
LA English
DT Article
AB Recent transcriptomic approaches focused on anti-viral immunity in molluscs lead to the assumption that the innate immune system, such as apoptosis, plays a crucial role against ostreid herpesvirus type 1 (OsHV-1), infecting Pacific cupped oyster, Crassostrea gigas. Apoptosis constitutes a major mechanism of anti-viral response by limiting viral spread and eliminating infected cells. In this way, an OsHV-1 challenge was performed and oysters were monitored at three times post injection to investigate viral infection and host response: 2h (early after viral injection in the adductor muscle), 24h (intermediate time), and 48h (just before first oyster mortality record). Virus infection, associated with high cumulative mortality rates (74% and 100%), was demonstrated in haemocytes by combining several detection techniques such as real-time PCR, real-time RT PCR, immunofluorescence assay, and transmission electron microscopy examination. High viral DNA amounts ranged from 5.46×104 to 3.68×105 DNA copies ng-1 of total DNA, were detected in dead oysters and an increase of viral transcripts was observed from 2, 24, and 48hpi for the five targeted OsHV-1 genes encoding three putative membrane proteins (ORFs 25, 41, and 72), a putative dUTPase (ORF 75), and a putative apoptosis inhibitor (ORF 87). Apoptosis was studied at molecular and cellular levels with an early marker (phosphatidyl-serine externalisation measured by flow cytometry and epifluorescence microscopy) and a later parameter (DNA fragmentation by terminal deoxynucleotidyltransferase-mediated dUTP nick end labeling assay (TUNEL)). The down-regulation of genes encoding proteins involved in the activation of the apoptotic pathway (TNF and caspase 3) and the up-regulation of genes encoding anti-apoptotic proteins (IAP-2, and Bcl-2) suggested an important anti-apoptosis phenomenon in haemocytes from OsHV-1 infected oysters at 24 and 48hpi. Additionally, more phosphatidyl-serines were externalized and more cells with DNA fragmentation were observed in haemocytes collected from artificial seawater injected oysters than in haemocytes collected from OsHV-1 infected oysters at 24 and 48hpi, suggesting an inhibition of the apoptotic process in presence of the virus. In conclusion, this study is the first to focus on C. gigas haemocytes, cells involved in the host immune defense, during an OsHV-1 challenge in controlled conditions by combining various and original approaches to investigate apoptosis at molecular and cellular levels. 
PY 2017
PD MAY
SO Plos One
SN 1932-6203
PU Public Library Science
VL 12
IS 5
UT 000401672400045
DI 10.1371/journal.pone.0177448
ID 49715
ER

EF