关键词: 斑马鱼, 转基因, 荧光激活细胞分选

Abstract: It was to optimize commonly used FACS protocols to apply to fluorescence-labeled neurons of zebrafish. lhx5:Kaede transgene line as an example was used to describe a successful protocol for isolating and expression-profiling live fluorescent-protein-labeled neurons from transgenic zebrafish brains. Numerous information of lhx5 labeling cells can be obtained by RNA profiling and would contribute to further studies. Results showed that on average, 6% of the single cell suspensions was keade-positive. The RNA extracted from selected cells was sufficient quality and quantity for microarray analysis. Microarray data showed the enrichment of both lhx5 and other molecular makers. It proved that this protocol for dissociating and sorting fluorescent-labeled neurons from central nervous system of zebrafish larve, is applicable for gene expression profiling of the target cell groups.

Key words: Zebrafish, Transgene, FACS