Rapid in vivo labeling of identified zebrafish neurons
Rapid in vivo labeling of identified zebrafish neurons
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DOI：
10.1002/gene.10120
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发表时间：
2002-11-01
期刊：
影响因子：
1.5
通讯作者：
Granato, M
中科院分区：
文献类型：
--
作者：
Downes, GB;Waterbury, JA;Granato, M
我们报告了一种简单而快速的方法来标记活斑马鱼胚胎中的单个神经元，并检查其基因表达谱。注射编码驱动GFP表达的α-微管蛋白启动子的质粒DNA导致含有有限数量的GFP阳性神经元的嵌合体胚胎。标记的神经元在其索马和轴突中表达GFP，为分析体内识别的神经元的寻路行为提供了机会。此外，仅存在一小部分GFP标记的神经元允许基于索马位置和轴突轨迹快速解剖学鉴定这些神经元。对注射胚胎的分析表明，大多数（如果不是全部的话）脊髓细胞类型和神经系统中其他地方的许多其他神经元细胞类型可以被GFP标记。最后，通过结合GFP标记的单个神经元与荧光原位杂交，我们证明了这种方法的潜力，阐明在单细胞分辨率的基因表达模式。(C)2002 Wiley-Liss，Inc.
We report a simple and rapid method to label individual neurons in live zebrafish embryos and to examine their gene expression profiles. Injection of plasmid DNA encoding an a-tubulin promotor driving GFP expression results in mosaic embryos containing a limited number of GFP-positive neurons. Labeled neurons express GFP in their soma and axon, providing the opportunity to analyze pathfinding behaviors of identified neurons in vivo. Moreover, the presence of only a small subset of GFP tagged neurons permits the rapid anatomical identification of these neurons based on soma position and axonal trajectory. Analysis of injected embryos reveals that most, if not all, spinal cord cell types and many other neuronal cell types elsewhere in the nervous system can be GFP tagged. Finally, by combining GFP labeling of individual neurons with fluorescent in situ hybridization, we demonstrate the potential of this method to elucidate gene expression patterns at single cell resolution. (C) 2002 Wiley-Liss, Inc.