Discovery of the Genes in Response to White Spot Syndrome Virus (WSSV) Infection in Fenneropenaeus chinensis Through cDNA Microarray

Abstract

We used microarray technology to study differentially expressed genes in white spot syndrome virus (WSSV)-infected shrimp. A total of 3136 cDNA targets, including 1578 unique genes from a cephalothorax cDNA library and 1536 cDNA clones from reverse and forward suppression subtractive hybridization (SSH) libraries of Fenneropenaeus chinensis, plus 14 negative and 8 blank control clones, were spotted onto a 18 x 18 mm area of NH(2)-modified glass slides. Gene expression patterns in the cephalothorax of shrimp at 6 h after WSSV injection and moribund shrimp naturally infected by WSSV were analyzed. A total of 105 elements on the arrays showed a similar regulation pattern in artificially infected shrimp and naturally infected moribund shrimp; parts of the results were confirmed by semiquantitative reverse transcriptase-polymerase chain reaction (RT-PCR). The up-regulated expression of immune-related genes, including heat shock proteins (HSP70 and HSP90), trehalose-phosphate synthase (TPS), ubiquitin C, and so forth, were observed when shrimp were challenged with WSSV. Genes including myosin LC2, ATP synthase A chain, and arginine kinase were found to be down-regulated after WSSV infection. The expression of housekeeping genes such as actin, elongation factor, and tubulin is not stable, and so these genes are not suitable as internal standards for semiquantitative RT-PCR when shrimp are challenged by WSSV. As a substitute, we found that triosephosphate isomerase (TPI) was an ideal candidate of interstandards in this situation.

Differentially expressed genes in Penaeus monodon hemocytes following infection with yellow head virus

“…The other commonly induced genes were calcium-dependent genes and transcription-and translation-related genes. However, in this study, YHV infection of P. monodon did not affect the expression level of several calcium-dependent genes including calnexin, calponin, calreticulin and calcium-sensitive chloride channel, in contrast to the reported up-regulation of calreticulin in WSSV-infected shrimp at http://bmbreports.org BMB reports both the mRNA and protein levels (3,18). In addition, no calcium-dependent protein was identified by comparative proteomic analysis between normal and YHV-infected shrimp (6).…”

“…Downregulation of several cell structure proteins and energy metabolism enzymes such as arginine kinase, actin, ATP synthase β-subunit and ATPases have been observed in WSSV-infected shrimp (3,17). In addition, reduction of the carbohydrate metabolic enzyme, 6-phosphogluconate dehydrogenase, was identified from comparative proteomic analysis of normal and YHV-infected shrimp.…”

“…β-Tubulin homologues were down-regulated in YHV infection, but were up-regulated in WSSV infection. Tubulin has long been known to be involved in the assembly and transport of virus particles (3,4). However, the response of β-tubulin to YHV infection and its functions in viral pathogenesis of WSSV and YHV are currently unknown.…”

“…Shrimp immunity can be studied on one level through molecular characterization of immune effectors and analysis of gene expression in response to a given pathogen. Viral responsive genes, including those encoding structural proteins, defense and homeostasis proteins, energy metabolism enzymes, transcription or translation proteins and proteins of unknown function, have been identified in white spot syndrome virus (WSSV)-challenged shrimp and WSSV-resistant shrimp (2)(3)(4). In contrast, the responses of shrimp to YHV infection at the molecular level are currently poorly described.…”

Gene Expression Patterns During the Larval Development of European Sea Bass (Dicentrarchus Labrax) by Microarray Analysis

“…There is indeed a consensus that regulation of gene expression is a fundamental mechanism underlying the complex processes of cell differentiation and morphogenesis during ontogenesis of an organism. Recently, microarrays have dramatically accelerated many types of investigations in marine organisms, been especially used for comparative genomic hybridization (Fish; Moriya et al, 2004 and2007) and for monitoring the expression levels of thousands of genes simultaneously in order to study the effects of certain treatments (Algae; Jamers et al, 2006), diseases (Crustaceans; Wang et al, 2006) and transgenesis manipulation (Mori et al, 2007) on transcriptome. Over the last two decades, studies based on whole-body or tissue-specific microarray approaches have investigated gene expression profiles during embryogenesis in several species (Drosophila, White et al, 1999;Xenopus, Baldessari et al, 2005;mouse, Tanaka et al, 2000) including fish (zebrafish, Ton et al, 2002;sea bream, Sarropoulou et al, 2005).…”