Gene expression profiling in gill tissues of White spot syndrome virus infected black tiger shrimp Penaeus monodon by DNA microarray
- 261 Downloads
White spot syndrome virus (WSSV) continues to be the most devastating viral pathogen infecting penaeid shrimp the world over. The genome of WSSV has been deciphered and characterized from three geographical isolates and significant progress has been made in developing various molecular diagnostic methods to detect the virus. However, the information on host immune gene response to WSSV pathogenesis is limited. Microarray analysis was carried out as an approach to analyse the gene expression in black tiger shrimp Penaeus monodon in response to WSSV infection. Gill tissues collected from the WSSV infected shrimp at 6, 24, 48 h and moribund stage were analysed for differential gene expression. Shrimp cDNAs of 40,059 unique sequences were considered for designing the microarray chip. The Cy3-labeled cRNA derived from healthy and WSSV-infected shrimp was subjected to hybridization with all the DNA spots in the microarray which revealed 8,633 and 11,147 as up- and down-regulated genes respectively at different time intervals post infection. The altered expression of these numerous genes represented diverse functions such as immune response, osmoregulation, apoptosis, nucleic acid binding, energy and metabolism, signal transduction, stress response and molting. The changes in gene expression profiles observed by microarray analysis provides molecular insights and framework of genes which are up- and down-regulated at different time intervals during WSSV infection in shrimp. The microarray data was validated by Real Time analysis of four differentially expressed genes involved in apoptosis (translationally controlled tumor protein, inhibitor of apoptosis protein, ubiquitin conjugated enzyme E2 and caspase) for gene expression levels. The role of apoptosis related genes in WSSV infected shrimp is discussed herein.
KeywordsMicroarray WSSV Penaeus monodon
Authors acknowledge Department of Biotechnology, Ministry of Science and Technology, India for funding this study under the project “Molecular studies on sequential pathogenesis of WSSV and defence mechanism in Penaeus monodon”.
- 6.Wang H, Lin A, Yii D, Chang Y, Kou G, Lo C. DNA microarrays of the White spot syndrome virus genome: genes expressed in the gills of infected shrimp. Mar Biotechnol. 2004;6:S106–11.Google Scholar
- 11.Robalino J, Almeida JS, McKillen D, Colglazier J, Trent HF III, Chen YA, Peck ME, Browdy CL, Chapman RW, Warr GW, Gross PS. Insights into the immune transcriptome of the shrimp Litopenaeus vannamei: tissue-specific expression profiles and transcriptomic responses to immune challenge. Physiol Genomics. 2007;29:44–56.PubMedCrossRefGoogle Scholar
- 14.Kou GH, Peng SE, Chiu YL, Lo CF. Tissue distribution of White spot syndrome virus (WSSV) in shrimp and crabs. In: Flegel TW, editor. Advances in shrimp biotechnology. Bangkok: National Center for Genetic Engineering and Biotechnology; 1998. p. 267–71.Google Scholar
- 15.Clavero-Salas A, Sotelo-Mundo RR, Gollas-Galván T, Hernández-López J, Beatriz Peregrino-Uriarte A, Muhlia-Almazán A, Yepiz-Plascencia G. Transcriptome analysis of gills from the white shrimp Litopenaeus vannamei infected with White spot syndrome virus. Fish Shellfish Immunol. 2007;23:459–72.PubMedCrossRefGoogle Scholar
- 27.Flegel TW, Pasharawipas T. Active viral accommodation: a new concept for crustacean response to viral pathogen. In: Flegel TW, editor. Advances in Shrimp Biotechnology. Bangkok: National Centre for Genetic Engineering and Biotechnology; 1998. p. 245–50.Google Scholar
- 34.Wang PH, Wan DH, Gu ZH, Qiu W, Chen YG, Weng SP, Yu XQ, He JG. Analysis of expression, cellular localization, and function of three inhibitors of apoptosis (IAPs) from Litopenaeus vannamei during WSSV infection and in regulation of antimicrobial peptide genes (AMPs). PLoS One. 2013;8(8):e72592.PubMedCentralPubMedCrossRefGoogle Scholar