Global analysis of hematopoietic and vascular endothelial gene expression by tissue specific microarray profiling in zebrafish
Program in Gene Function and Expression; Program in Molecular Medicine
Animals; Animals, Genetically Modified; Branchial Region; Cell Separation; Endothelium, Vascular; Gene Expression; Green Fluorescent Proteins; Hematopoietic System; Oligonucleotide Array Sequence Analysis; Zebrafish; Zebrafish Proteins
Life Sciences | Medicine and Health Sciences
In this study, we utilize fluorescent activated cell sorting (FACS) of cells from transgenic zebrafish coupled with microarray analysis to globally analyze expression of cell type specific genes. We find that it is possible to isolate cell populations from Tg(fli1:egfp)(y1) zebrafish embryos that are enriched in vascular, hematopoietic and pharyngeal arch cell types. Microarray analysis of GFP+ versus GFP- cells isolated from Tg(fli1:egfp)(y1) embryos identifies genes expressed in hematopoietic, vascular and pharyngeal arch tissue, consistent with the expression of the fli1:egfp transgene in these cell types. Comparison of expression profiles from GFP+ cells isolated from embryos at two different time points reveals that genes expressed in different fli1+ cell types display distinct temporal expression profiles. We also demonstrate the utility of this approach for gene discovery by identifying numerous previously uncharacterized genes that we find are expressed in fli1:egfp-positive cells, including new markers of blood, endothelial and pharyngeal arch cell types. In parallel, we have developed a database to allow easy access to both our microarray and in situ results. Our results demonstrate that this is a robust approach for identification of cell type specific genes as well as for global analysis of cell type specific gene expression in zebrafish embryos.
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Citation: Dev Biol. 2006 Nov 15;299(2):551-62. Epub 2006 Aug 10. Link to article on publisher's site