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Ecdysteroid-responsive genes: E75 and Retinoid X Receptor (RXR), essential molt-regulating genes in the shrimp, Fenneropenaeus chinensis
其他题名Ecdysteroid-responsive genes: E75 and Retinoid X Receptor (RXR), essential molt-regulating genes in the shrimp, Fenneropenaeus chinensis
JOSEPRIYA THANKARAJ ALPHONSAL
学位类型博士
2009-01-05
学位授予单位中国科学院海洋研究所
学位授予地点海洋研究所
关键词蜕皮激素响应基因 E75和retinoid X受体 蜕皮 中国明对虾
摘要The study presented in this thesis is an attempt to explore the potential involvement of two important molt-regulating genes: generally called ecdysone responsive genes, E75 and RXR in the molting process of Chinese shrimp, Fenneropenaeus chinensis. Full-length cDNAs encoding FcE75 and FcRXR were obtained using PCR technique. Genomic structure of FcRXR with 7 introns depicts the existence of several isoforms in shrimp and we identified two, namely RXR-1 and RXR-2. Quantitative Real time PCR confirmed the prominent expression of FcE75 and FcRXR mRNAs in the late pre-molt period (D3) of shrimp. Moreover, a marked tissue-specific difference in the expression of FcE75 and FcRXR transcripts were also seen which revealed that the expression of FcE75 could be regulated in a tissue-specific manner. Interestingly, the application of double strand RNA into the shrimp efficiently decreased the FcE75 and FcRXR expression levels. Observations suggested that FcE75 silencing in vivo completely arrested the molting process and caused death of the shrimp. Analysis of the animals which failed to molt showed defective epidermal retraction, poor development of setae and new cuticle. On the other hand, silencing of both FcE75 & FcRXR in turn leads to considerable reduction in the levels of chitinase and chitinase 1 gene. Further, FcE75 silenced samples revealed reduced level of FcRXR expression and vice versa. These results indicate that FcE75 & FcRXR exert a prominent function during the molting process. We conclude that the ecdysone responsive genes FcE75 and FcRXR are essential for proper molting and survival of shrimp. This is the first report elucidating the function of these genes in the molting of decapod crustaceans.
其他摘要本论文主要研究两种重要的调节蜕皮过程的基因—蜕皮激素效应基因E75和RXR在中国明对虾蜕皮中的作用。利用RT-PCR和RACE技术获得了编码FcE75和FcRXR的全长cDNA序列。FcRXR包含7个内含子,在对虾中存在不同的异形体,命名为RXR-1和RXR-2。应用荧光实时定量PCR分析表明FcE75和FcRXR基因在中国明对虾蜕皮前期(D3)其转录表达量明显上调。另外,FcE75和FcRXR基因在不同组织中的转录表达存在明显的差异。利用FcE75和FcRXR基因的双链RNA注射对虾能有效降低FcE75和FcRXR的表达水平。FcE75和FcRXR的体内沉默完全抑制了对虾的蜕皮过程,并且引起对虾的死亡。对不能正常蜕皮个体进行观察的结果表明,FcE75沉默的对虾,其上皮的收缩、新的刚毛及新表皮的形成均收到限制。在FcE75双链RNA沉默后的对虾中,我们检测了与蜕皮相关的一些效应因子,如chitinase等的转录,发现这些效应因子的转录明显受到抑制,说明FcE75和FcRXR在蜕皮过程中起到非常重要的作用。本论文首次阐明了这些基因在十足目甲壳动物蜕皮过程中的功能。
页数127
语种中文
文献类型学位论文
条目标识符http://ir.qdio.ac.cn/handle/337002/801
专题海洋环流与波动重点实验室
推荐引用方式
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JOSEPRIYA THANKARAJ ALPHONSAL. Ecdysteroid-responsive genes: E75 and Retinoid X Receptor (RXR), essential molt-regulating genes in the shrimp, Fenneropenaeus chinensis[D]. 海洋研究所. 中国科学院海洋研究所,2009.
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