Shekhar M S, Gomathi A, Gopikrishna G, Ponniah A G
Genetics and Biotechnology Unit, Central Institute of Brackishwater Aquaculture, 75 Santhome High Road, R.A. Puram, Chennai, 600028 India.
Virusdisease. 2015 Jun;26(1-2):9-18. doi: 10.1007/s13337-014-0243-7. Epub 2015 Feb 10.
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.
白斑综合征病毒(WSSV)仍然是全球感染对虾的最具毁灭性的病毒病原体。已对来自三个地理分离株的WSSV基因组进行了解码和表征,并且在开发各种检测该病毒的分子诊断方法方面取得了重大进展。然而,关于宿主免疫基因对WSSV发病机制反应的信息有限。进行了微阵列分析,以分析黑虎虾凡纳滨对虾在感染WSSV后的基因表达。对在6、24、48小时和濒死阶段从感染WSSV的虾中收集的鳃组织进行差异基因表达分析。考虑了40,059个独特序列的虾cDNA用于设计微阵列芯片。将来自健康和感染WSSV的虾的Cy3标记的cRNA与微阵列中的所有DNA斑点进行杂交,结果显示在感染后的不同时间间隔分别有8,633个和11,147个基因上调和下调。这些众多基因的表达改变代表了多种功能,如免疫反应、渗透调节、细胞凋亡、核酸结合、能量和代谢、信号转导、应激反应和蜕皮。通过微阵列分析观察到的基因表达谱变化提供了分子见解以及在对虾感染WSSV期间不同时间间隔上调和下调的基因框架。通过对四个参与细胞凋亡的差异表达基因(翻译控制肿瘤蛋白、凋亡抑制蛋白泛素缀合酶E2和半胱天冬酶)的基因表达水平进行实时分析,验证了微阵列数据。本文讨论了细胞凋亡相关基因在感染WSSV的对虾中的作用。