Goto Rie, Saito Taiju, Matsubara Takahiro, Yamaha Etsuro
South Ehime Fisheries Research Center, Ehime University, Minamiuwagun, Ehime, Japan.
Nanae Fresh Water Laboratory, Field Science Center for Northern Biosphere, Hokkaido University, Kameda, Hokkaido, Japan.
Methods Mol Biol. 2019;1874:475-487. doi: 10.1007/978-1-4939-8831-0_27.
Microinjection is a powerful tool for studying embryonic development and analyzing gene functions in fish. This technique was first applied to model species of fish such as zebrafish and medaka whose egg chorions could be removed or softened before microinjection. Recent progress in genome editing using TALEN and CRISPR has opened the opportunity to analyze gene functions in a much wider range of fish including those important to marine aquaculture. Therefore, application of the microinjection technique is also required in these species. However, the characteristics of fish eggs vary widely between species and several technical difficulties need to be overcome in order to use microinjection in a wider range of species. To obtain consistent results with microinjection, an optimal method has to be developed for each target species. In this chapter, we describe the physical characteristics of the eggs of fish species that have been used in microinjection experiments in our laboratory and detail the microinjection system we developed for fish eggs with a hard chorion, such as those of marine species.
显微注射是研究鱼类胚胎发育和分析基因功能的有力工具。该技术最初应用于斑马鱼和青鳉等鱼类模式物种，这些物种的卵膜在显微注射前可以去除或软化。利用TALEN和CRISPR进行基因组编辑的最新进展为在更广泛的鱼类中分析基因功能提供了机会，包括那些对海水养殖很重要的鱼类。因此，这些物种也需要应用显微注射技术。然而，不同鱼类的鱼卵特性差异很大，为了在更广泛的物种中使用显微注射技术，需要克服一些技术难题。为了通过显微注射获得一致的结果，必须为每个目标物种开发一种最佳方法。在本章中，我们描述了我们实验室用于显微注射实验的鱼类物种的鱼卵物理特性，并详细介绍了我们为具有硬卵膜的鱼卵（如海洋物种的鱼卵）开发的显微注射系统。
