Okazaki Institute for Integrative Bioscience, Japan.
Ecotoxicology. 2010 Mar;19(3):589-92. doi: 10.1007/s10646-010-0460-9. Epub 2010 Jan 23.
Daphnids inhabit a diverse array of aquatic environments and they are a good model for understanding response and adaptation to environmental changes and they have been used one of standard organisms in ecotoxicology. Recent progress of genomics changed the tools for analyzing responses of daphnids, because gene expression changes can be observed before the emergence of prominent adverse effect such as immobility of the organism. Thus understanding of biological changes from gene expression level can be one of the sensitive tools for the evaluation of environmental response of organisms. However, there was no technique for genetic manipulation in daphnids. Hence, we have developed a gene introduction technique based on electroporation. There are two critical points for the successful introduction of foreign DNA into D. magna. (1) Injection of DNA into blood stream. (2) Usage of very low voltage for the electroporation. The injected DNA containing green fluorescent protein (GFP) could be introduced daphnids and the expression of GFP could be detected in living daphnids. This is the first report of gene introduction to daphnids and, together with the emerging genome sequences, will be useful for the expanding our use of daphnid in ecotoxicology.
枝角类栖息于多种多样的水生环境中，是研究环境变化响应和适应的良好模式生物，也是生态毒理学中标准的实验生物之一。基因组学的最新进展改变了分析枝角类响应的工具，因为在出现明显的不利影响（如生物体的不动性）之前，可以观察到基因表达的变化。因此，从基因表达水平理解生物学变化可以成为评估生物体环境响应的敏感工具之一。然而，在枝角类中没有基因操作技术。因此，我们开发了一种基于电穿孔的基因导入技术。将外源 DNA 成功导入大型溞有两个关键要点。(1) 将 DNA 注射到血流中。(2) 电穿孔时使用非常低的电压。含有绿色荧光蛋白 (GFP) 的注射 DNA 可以被导入大型溞，并且可以在活体大型溞中检测到 GFP 的表达。这是首次将基因导入大型溞的报道，与新兴的基因组序列一起，将有助于扩大大型溞在生态毒理学中的应用。
