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):2055. doi: 10.3791/2055.
Medaka is a small egg-laying freshwater fish that allows both genetic and embryological analyses and is one of the three vertebrate model organisms in which genome-wide phenotype-driven mutant screens were carried out (1). 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 the vertebrate genome functions. Manipulation of medaka embryos, such as dechorionation, mounting embryos for imaging and cell transplantation, are key procedures to work on both medaka and zebrafish in a laboratory. Cell transplantation examines cell autonomy of medaka mutations. Chimeras are generated by transplanting labeled cells from donor embryos into unlabeled recipient embryos. Donor cells can be transplanted to specific areas of the recipient embryos based on the fate maps (3) so that clones from transplanted cells can be integrated in the tissue of interest during development. Due to the hard chorion and soft embryos, manipulation of medaka embryos is more involved than in zebrafish. In this video, we show detailed procedures to manipulate medaka embryos.
青鳉是一种小型的产卵淡水鱼，可用于遗传和胚胎学分析，是进行全基因组表型驱动突变筛选的三种脊椎动物模式生物之一（1）。青鳉和斑马鱼相关基因功能重叠的差异使得能够鉴定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。对青鳉胚胎进行操作，如去绒毛膜、将胚胎固定用于成像和细胞移植，是在实验室中对青鳉和斑马鱼进行研究的关键步骤。细胞移植可检测青鳉突变的细胞自主性。通过将来自供体胚胎的标记细胞移植到未标记的受体胚胎中来产生嵌合体。根据命运图谱（3），可以将供体细胞移植到受体胚胎的特定区域，以便移植细胞的克隆在发育过程中能够整合到感兴趣的组织中。由于青鳉胚胎的绒毛膜坚硬而胚胎柔软，对青鳉胚胎的操作比斑马鱼更为复杂。在本视频中，我们展示了操作青鳉胚胎的详细步骤。
