Rapid in vivo labeling of identified zebrafish neurons.
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 alpha-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.
Downes
Gerald B
GB
Department of Cell and Developmental Biology, University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania 19104-6058, USA.
Waterbury
Julie A
JA
Granato
Michael
M
D000818:Animals; D000906:Antibodies / Q000276:immunology; D017930:Genes, Reporter / Q000276:immunology; D017404:In Situ Hybridization, Fluorescence; D009474:Neurons / Q000166:cytology* / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D013116:Spinal Cord / Q000166:cytology / Q000378:metabolism; D013194:Staining and Labeling / Q000379:methods*; D014404:Tubulin / Q000378:metabolism*; D015027:Zebrafish / Q000378:metabolism*
D000906:Antibodies; D011993:Recombinant Fusion Proteins; D014404:Tubulin