National Center for Genetic Engineering and Biotechnology (BIOTEC), National Science and Technology Development Agency (NSTDA), Klong 1, Klong Luang, Pathumthani 12120, Thailand.
BMC Mol Biol. 2011 Apr 12;12:15. doi: 10.1186/1471-2199-12-15.
Poor reproductive maturation in captive male broodstock of the black tiger shrimp (Penaeus monodon) is one of the serious problems to the farming industries. Without genome sequence, EST libraries of P. monodon were previously constructed to identify transcripts with important biological functions. In this study, a new version of cDNA microarray, UniShrimpChip, was constructed from the Peneaus monodon EST libraries of 12 tissues, containing 5,568 non-redundant cDNA clones from 10,536 unique cDNA in the P. monodon EST database. UniShrimpChip was used to study testicular development by comparing gene expression levels of wild brooders from the West and East coasts of Thailand and domesticated brooders with different ages (10-, 14-, 18-month-old).
The overall gene expression patterns from the microarray experiments revealed distinct transcriptomic patterns between the wild and domesticated groups. Moreover, differentially expressed genes from the microarray comparisons were identified, and the expression patterns of eight selected transcripts were subsequently confirmed by reverse-transcriptase quantitative PCR (RT-qPCR). Among these, expression levels of six subunits (CSN2, 4, 5, 6, 7a, and 8) of the COP9 signalosome (CSN) gene family in wild and different ages of domesticated brooders were examined by RT-qPCR. Among the six subunits, CSN5 and CSN6 were most highly expressed in wild brooders and least expressed in the 18-month-old domesticated group; therefore, their full-length cDNA sequences were characterized.
This study is the first report to employ cDNA microarray to study testicular development in the black tiger shrimp. We show that there are obvious differences between the wild and domesticated shrimp at the transcriptomic level. Furthermore, our study is the first to investigate the feasibility that the CSN gene family might have involved in reproduction and development of this economically important species.
人工养殖的斑节对虾（Penaeus monodon）亲虾繁殖成熟度低是养殖行业面临的严重问题之一。由于缺乏基因组序列，此前曾构建了斑节对虾 EST 文库，以鉴定具有重要生物学功能的转录本。在这项研究中，从泰国西海岸和东海岸的野生亲虾和不同年龄（10、14、18 月龄）的人工养殖亲虾的 12 种组织的斑节对虾 EST 文库中构建了一个新的 cDNA 微阵列 UniShrimpChip，该微阵列包含来自斑节对虾 EST 数据库中的 10536 个独特 cDNA 的 5568 个非冗余 cDNA 克隆。UniShrimpChip 用于通过比较来自泰国西海岸和东海岸的野生亲虾和不同年龄（10、14、18 月龄）的人工养殖亲虾的睾丸发育过程中的基因表达水平来研究睾丸发育。
微阵列实验的整体基因表达模式揭示了野生和家养群体之间明显的转录组模式。此外，还鉴定了微阵列比较中差异表达的基因，并通过逆转录定量 PCR（RT-qPCR）随后验证了 8 个选定转录本的表达模式。其中，在野生和不同年龄的家养亲虾中，COX9 信号小体（CSN）基因家族的 6 个亚基（CSN2、4、5、6、7a 和 8）的表达水平通过 RT-qPCR 进行了检测。在这 6 个亚基中，CSN5 和 CSN6 在野生亲虾中表达最高，在 18 月龄的家养组中表达最低；因此，对其全长 cDNA 序列进行了表征。
本研究首次采用 cDNA 微阵列研究斑节对虾的睾丸发育。我们表明，在转录组水平上，野生和家养亲虾之间存在明显差异。此外，我们的研究首次调查了 CSN 基因家族可能参与这种经济重要物种的繁殖和发育的可行性。
