Suppression subtraction hybridization (SSH) and macroarray techniques reveal differential gene expression profiles in brain of sea bream infected with nodavirus

Abstract

Despite of the impact that viruses have on aquatic organisms, relatively little is known on how fish fight against these infections. In this work, the brain gene expression pattern of sea bream (Sparus aurata) in response to nodavirus infection was investigated. We used the SSH (Supression Subtractive Hybridization) method to generate a subtracted cDNA library enriched with gene transcripts differentially expressed after one day post infection. Some of the ESTs from the infected tissues fell in gene categories related to stress and immune responses. For the reverse library (ESTs expressed in controls compared with infected tissues) the most abundant transcripts were of ribosomal and mitochondrial nature. Several ESTs potentially induced by virus exposure were selected for in vivo expression studies. We observed a clear difference in expression between infected and control samples for two candidate genes, ubiquitin conjugating enzyme 7 interacting protein, which seems to play an important role in apoptosis and the interferon induced protein with helicase C domain 1 (mda-5) that contributes to apoptosis and regulates the type I IFN production, a key molecule of the antiviral innate response in most organisms.

Characterization of the BPI-like gene from a subtracted cDNA library of large yellow croaker (Pseudosciaena crocea) and induced expression by formalin-inactivated Vibrio alginolyticus and Nocardia seriolae vaccine challenges

“…These include Oncorhyncus mykiss injected with Vibrio anguillarum [2]; P. crocea injected with polyI:C [36]; Sparus aurata injected with Nodavirus [37]; Paralichthys olivaceus injected with PG [8]; Cyprinus carpio injected with PG, LPS and Con-A [9,10]; and Epinephelus awoara injected with LPS [11]. In this paper, a subtracted cDNA library from the head kidney of the large yellow croaker stimulated with PG has been constructed using SSH technology, and a novel Pc-BPI-L gene in large yellow croaker has been thus identified.…”

Identification and molecular cloning of Atlantic cod (Gadus morhua) activating transcription factor 3 (ATF3) transcript and its induction in spleen following intraperitoneal polyriboinosinic polyribocytidylic acid injection

“…cDNA libraries, such as suppression subtractive hybridization (SSH) libraries, have been extensively used in functional genomics projects of many fish species (e.g [11,12]. including Atlantic cod (Gadus morhua) (e.g [13e15].).…”

Nodavirus Infection of Sea Bass (Dicentrarchus labrax) Induces Up-Regulation of Galectin-1 Expression with Potential Anti-Inflammatory Activity

“…The percentages of the significant ESTs obtained were higher than for cDNA or subtracted libraries after experimental infection or immune stimulation of Japanese flounder (Paralichthys olivaceus) (30), carp (Cyprinus carpio) (31), salmon (S. salar) (32), and gilthead seabream (Sparus aurata) (33). Within the genes related to the immune system category, however, we identified genes that had already been described in those fish libraries (Table I) and are critical components in the mammalian immune response such as galectins (34,35), Cd209e (35), C-type lectins (31,32,34,36,37), pentraxins (32,38), MHC class II and heat-shock protein 90 (30,32,35), ␤ 2 -microglobulin, Ig H and L chains (30,32), and Tcomplex protein (32).…”