ScienceAsia 33 (2007): 165-174 |doi: 10.2306/scienceasia1513-1874.2007.33.165

Analysis of Gene Expression in Haemocytes of Shrimp Penaeus monodon Challenged with White Spot Syndrome Virus by cDNA Microarray

Ratree Wongpanya^a, Takashi Aoki^b, Ikuo Hirono^b, Motoshige Yasuike^b and Anchalee Tassanakajon^a*

 
ABSTRACT: A cDNA microarray technique was employed to identify differentially-expressed genes in the black tiger shrimp (Penaeus monodon) challenged with white spot syndrome virus (WSSV). The cDNA chip was composed of 718 unique genes from P. monodon and 308 unique genes from the kuruma shrimp (Marsupenaeus japonicus). The cDNA arrays were hybridized with Cy3/Cy5-labeled probes generated from the haemocyte RNA of P. monodon injected with WSSV. The gene expression profile was determined at different time points after injection. The results showed differential expression in haemocytes of WSSV-challenged P. monodon. The number of up- and down-regulated genes was highest at 24 h after WSSV injection. The pathogen invasion resulted in up-regulation of ribosomal proteins, indicating an increase in protein synthesis. The expressions of several immune-related genes also changed. The mRNA expressions of three highly responsive genes (calmodulin, tubulin and asialoglycoprotein receptor) in response to shrimp pathogens, WSSV and the luminescence bacteria Vibrio harveyi, were verified by real-time PCR and confirmed the involvement of these genes in shrimp defense. Our results suggested the WSSV infection altered the expression of a wide range of cellular genes. The microarray analysis identified several clones with unknown function that were upregulated by WSSV infection. The roles of these clones in shrimp immune response need to be further investigated.

KEYWORDS: microarray, shrimp, real-time PCR, gene expression,WSSV.

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