Pratt T, Sharp L, Nichols J, Price D J, Mason J O
Department of Biomedical Sciences, Centre for Developmental Biology, The University of Edinburgh, Hugh Robson Building, George Square, Edinburgh, EH8 9XD, United Kingdom.
Dev Biol. 2000 Dec 1;228(1):19-28. doi: 10.1006/dbio.2000.9935.
We have generated embryonic stem (ES) cells and transgenic mice carrying a tau-tagged green fluorescent protein (GFP) transgene under the control of a powerful promoter active in all cell types including those of the central nervous system. GFP requires no substrate and can be detected in fixed or living cells so is an attractive genetic marker. Tau-tagged GFP labels subcellular structures, including axons and the mitotic machinery, by binding the GFP to microtubules. This allows cell morphology to be visualized in exquisite detail. We test the application of cells derived from these mice in several types of cell-mixing experiments and demonstrate that the morphology of tau-GFP-expressing cells can be readily visualized after they have integrated into unlabeled host cells or tissues. We anticipate that these ES cells and transgenic mice will prove a novel and powerful tool for a wide variety of applications including the development of neural transplantation technologies in animal models and fundamental research into axon pathfinding mechanisms. A major advantage of the tau-GFP label is that it can be detected in living cells and labeled cells and their processes can be identified and subjected to a variety of manipulations such as electrophysiological cell recording.
我们已生成胚胎干细胞（ES细胞）和转基因小鼠，这些小鼠携带一个在包括中枢神经系统细胞在内的所有细胞类型中都有活性的强大启动子控制下的tau标记绿色荧光蛋白（GFP）转基因。GFP无需底物，可在固定或活细胞中检测到，因此是一种有吸引力的遗传标记。Tau标记的GFP通过将GFP与微管结合来标记亚细胞结构，包括轴突和有丝分裂机制。这使得细胞形态能够以极高的细节被可视化。我们在几种细胞混合实验中测试了源自这些小鼠的细胞的应用，并证明表达tau-GFP的细胞在整合到未标记的宿主细胞或组织后，其形态能够很容易地被可视化。我们预计，这些ES细胞和转基因小鼠将被证明是一种新颖且强大的工具，可用于多种应用，包括动物模型中神经移植技术的开发以及轴突寻路机制的基础研究。tau-GFP标记的一个主要优点是它可以在活细胞中被检测到，并且标记的细胞及其突起可以被识别并进行各种操作，如电生理细胞记录。
