title: Proteomic analysis of hemolymph from poly(I:C)-stimulated Crassostrea gigas.
abstract: Synthetic double stranded RNA (Poly(I:C)) injection of Crassostrea gigas results in a systemic antiviral response involving many evolutionary conserved antiviral effectors (ISGs). Compared to mammals, the timing of C. gigas ISG expression to viral or poly(I:C) injection is delayed (>12 h p.i.). It could be interpreted that a cytokine is responsible for the systemic, but delayed expression of C. gigas ISGs. We therefore analysed the acellular fraction of C. gigas hemolymph by two-dimensional electrophoresis (2-DE) to identify hemolymph proteins induced by poly(I:C). Poly(I:C) injection increased the relative intensity of four protein spots. These protein spots were identified by tandem mass spectrometry (LC-MS/MS) as a small heat shock protein (sHSP), poly(I:C)-inducible protein 1 (PIP1) and two isoforms of C1q-domain containing protein (C1qDC). RT-qPCR analysis confirmed that the genes encoding these proteins are induced in hemocytes of C. gigas injected with poly(I:C) (p < 0.05). Proteomic data from this experiment corroborates previous microarray and whole transcriptome studies that have reported up-regulation of C1qDC and sHSP during mass mortality events among farmed oysters.
authors.affiliation: Department of Biological Sciences, Macquarie University, Sydney, NSW, Australia; Sydney Institute of Marine Science, Chowder Bay, Mosman, Sydney, NSW, Australia. Electronic address: tim.green@mq.edu.au.
authors.forename: Timothy J
authors.initials: TJ
authors.lastname: Green
authors.affiliation: Department of Human Physiology and Centre for Neuroscience, Flinders University, Adelaide, SA, Australia.
authors.forename: Timothy
authors.initials: T
authors.lastname: Chataway
authors.affiliation: Department of Biological Sciences, Macquarie University, Sydney, NSW, Australia; Sydney Institute of Marine Science, Chowder Bay, Mosman, Sydney, NSW, Australia.
authors.forename: Aroon R
authors.initials: AR
authors.lastname: Melwani
authors.affiliation: Department of Biological Sciences, Macquarie University, Sydney, NSW, Australia; Sydney Institute of Marine Science, Chowder Bay, Mosman, Sydney, NSW, Australia.
authors.forename: David A
authors.initials: DA
authors.lastname: Raftos
mesh_terms: D000818:Animals; D000998:Antiviral Agents / Q000494:pharmacology; D060829:Arthropod Proteins / Q000378:metabolism*; D002352:Carrier Proteins / Q000378:metabolism; D049896:Crassostrea / Q000378:metabolism*; D050888:Heat-Shock Proteins, Small / Q000378:metabolism; D006434:Hemocytes / Q000378:metabolism; D006458:Hemolymph / Q000378:metabolism*; D011070:Poly I-C / Q000494:pharmacology; D040901:Proteomics
chemical_list: D000998:Antiviral Agents; D060829:Arthropod Proteins; D002352:Carrier Proteins; D050888:Heat-Shock Proteins, Small; D011070:Poly I-C
keywords: Antiviral; C1q-domain containing protein; Crassostrea; Two-dimensional electrophoresis; poly(I:C)