Division of Molecular and Developmental Biology, National Institute of Genetics, Department of Genetics, Graduate University for Advanced Studies (SOKENDAI), Mishima, Shizuoka, Japan.
Dev Dyn. 2010 Oct;239(10):2695-9. doi: 10.1002/dvdy.22400.
The microtubule (MT) cytoskeleton plays crucial roles in brain development by regulating the proliferation of neuronal progenitor cells, neuronal migration and axon guidance. Methods for monitoring MT in the intact brain, however, have been limited in vertebrates. Here, we report a transgenic zebrafish line for monitoring MT in vivo. This reporter line carries a transgene encoding the green fluorescent protein (GFP) -tagged tubulin gene linked to the upstream activating sequence (UAS), the recognition sequence of the yeast Gal4 transcriptional activator. By crossing this reporter line with appropriate transgenic Gal4 driver lines, we induced the GFP-tagged tubulin in various cell types from the embryonic stages to the adult stage. In larvae expressing the modified tubulin, individual MT filaments and other MT structures, including the mitotic spindles in proliferating neuronal progenitor cells, were clearly visualized. Therefore, the transgenic UAS reporter line should be useful for directly monitoring MTs in the intact brain.
微管（MT）细胞骨架通过调节神经元祖细胞的增殖、神经元迁移和轴突导向，在大脑发育中发挥着关键作用。然而，在脊椎动物中，监测完整大脑中的 MT 的方法一直受到限制。在这里，我们报告了一种用于体内监测 MT 的转基因斑马鱼品系。该报告品系携带一个转基因组，该转基因组编码与上游激活序列（UAS）连接的 GFP 标记的微管蛋白基因，UAS 是酵母 Gal4 转录激活因子的识别序列。通过将该报告系与适当的转基因 Gal4 驱动系杂交，我们在从胚胎期到成年期的各种细胞类型中诱导 GFP 标记的微管蛋白。在表达修饰后的微管蛋白的幼虫中，可以清楚地观察到单个 MT 丝和其他 MT 结构，包括增殖的神经元祖细胞中的有丝分裂纺锤体。因此，该转基因 UAS 报告系应该可用于直接监测完整大脑中的 MT。
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