Yoshida Mika, Macklin Wendy B
Department of Neurosciences, Lerner Research Institute, The Cleveland Clinic Foundation,Cleveland, Ohio 44195, USA.
J Neurosci Res. 2005 Jul 1;81(1):1-8. doi: 10.1002/jnr.20516.
Green fluorescent protein (GFP) transgenic zebrafish technology has been employed to directly visualize and analyze dynamic developmental processes, such as cell migration and morphogenesis. Stable transgenic zebrafish that express GFP in oligodendrocytes can be a valuable tool to visualize complex myelination processes in vivo, as well as to conduct rapid mutagenesis screens for defective myelination mutants. We investigated whether two myelin gene promoters, the zebrafish P0 promoter and the mouse proteolipid protein (PLP) promoter, drive GFP expression in zebrafish oligodendrocytes. Transiently, both promoters drive enhanced GFP (EGFP) expression in morphologically identifiable oligodendrocytes, premyelinating oligodendrocytes, and possible oligodendrocyte precursors. We have established a stable transgenic zebrafish line, tg(plp:EGFP) zebrafish, at the F1 generation, which expresses enhanced GFP (EGFP) driven by the mouse PLP promoter. In this transgenic line, EGFP-expressing cells are visually detectable around 24-hr postfertilization (hpf), and later at 54 hpf, these cells start exhibiting the clear morphologic characteristics of oligodendrocytes. Shortly afterward, EGFP-expressing oligodendrocytes establish a ventral dominant distribution pattern throughout the central nervous system. This transgenic zebrafish line is likely to serve as a useful tool, in which normal myelination as well as abnormal myelination can be recorded under time-lapse confocal microscopy. Furthermore, it has the potential to greatly facilitate mutagenesis screening for novel dysmyelinating mutants.
绿色荧光蛋白（GFP）转基因斑马鱼技术已被用于直接观察和分析动态发育过程，如细胞迁移和形态发生。在少突胶质细胞中表达GFP的稳定转基因斑马鱼可成为一种有价值的工具，用于在体内观察复杂的髓鞘形成过程，以及对髓鞘形成缺陷突变体进行快速诱变筛选。我们研究了两种髓鞘基因启动子，即斑马鱼P0启动子和小鼠蛋白脂蛋白（PLP）启动子，是否能驱动GFP在斑马鱼少突胶质细胞中表达。短暂地，这两种启动子均能在形态上可识别的少突胶质细胞、前髓鞘形成少突胶质细胞以及可能的少突胶质细胞前体中驱动增强型GFP（EGFP）表达。我们在F1代建立了一个稳定的转基因斑马鱼品系，即tg(plp:EGFP)斑马鱼，它表达由小鼠PLP启动子驱动的增强型GFP（EGFP）。在这个转基因品系中，受精后约24小时（hpf）可在视觉上检测到表达EGFP的细胞，随后在54 hpf时，这些细胞开始表现出少突胶质细胞清晰的形态特征。不久之后，表达EGFP的少突胶质细胞在整个中枢神经系统中建立起腹侧优势分布模式。这个转基因斑马鱼品系可能会成为一种有用的工具，通过延时共聚焦显微镜可以记录正常和异常的髓鞘形成过程。此外，它有潜力极大地促进对新型髓鞘形成障碍突变体的诱变筛选。
