Porazinski Sean R, Wang Huijia, Furutani-Seiki Makoto
Centre for Regenerative Medicine, Department of Biology and Biochemistry, University of Bath.
J Vis Exp. 2010 Dec 22(46):1937. doi: 10.3791/1937.
In this video, we demonstrate the technique of microinjection into one-cell stage medaka embryos. Medaka is a small egg-laying freshwater fish that allows both genetic and embryological analyses and is one of the vertebrate model organisms in which genome-wide phenotype-driven mutant screens were carried out (1), as in zebrafish and the mouse. Divergence of functional overlap of related genes between medaka and zebrafish allows identification of novel phenotypes that are unidentifiable in a single species (2), thus medaka and zebrafish are complementary for genetic dissection of vertebrate genome functions. To take advantage of medaka fish whose embryos are transparent and develop externally, microinjection is an essential technique to inject cell-tracers for labeling cells, mRNAs or anti-sense oligonucleotides for over-expressing and knocking-down genes of interest, and DNAs for making transgenic lines.
在本视频中，我们展示了对单细胞期青鳉胚胎进行显微注射的技术。青鳉是一种小型的产卵淡水鱼，可用于遗传和胚胎学分析，并且是进行全基因组表型驱动突变体筛选的脊椎动物模式生物之一（1），如同斑马鱼和小鼠。青鳉和斑马鱼相关基因功能重叠的差异使得能够鉴定出在单一物种中无法识别的新表型（2），因此青鳉和斑马鱼在脊椎动物基因组功能的遗传剖析方面具有互补性。为了利用胚胎透明且在体外发育的青鳉鱼，显微注射是一种必不可少的技术，可用于注射细胞追踪剂以标记细胞、注射mRNA或反义寡核苷酸以过表达和敲低感兴趣的基因，以及注射DNA以构建转基因品系。
