Dynamic visualization of nervous system in live Drosophila.
We have constructed transgenic Drosophila melanogaster lines that express green fluorescent protein (GFP) exclusively in the nervous system. Expression is controlled with transcriptional regulatory elements present in the 5' flanking DNA of the Drosophila Na(+), K(+)-ATPase beta-subunit gene Nervana2 (Nrv2). This regulatory DNA is fused to the yeast transcriptional activator GAL4, which binds specifically to a sequence motif termed the UAS (upstream activating sequence). Drosophila lines carrying Nrv2-GAL4 transgenes have been genetically recombined with UAS-GFP (S65T) transgenes (Nrv2-GAL4+UAS-GFP) inserted on the same chromosomes. We observe strong nervous system-specific fluorescence in embryos, larvae, pupae, and adults. The GFP fluorescence is sufficiently bright to allow dynamic imaging of the nervous system at all of these developmental stages directly through the cuticle of live Drosophila. These lines provide an unprecedented view of the nervous system in living animals and will be valuable tools for investigating a number of developmental, physiological, and genetic neurobiological problems.
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Division of Neuroscience, Beckman Research Institute of the City of Hope, 1450 East Duarte Road, Duarte, CA 91010, USA.
Xu
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Salvaterra
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D000818:Animals; D030801:Animals, Genetically Modified; D002460:Cell Line; D004268:DNA-Binding Proteins; D029721:Drosophila Proteins; D004331:Drosophila melanogaster / Q000254:growth & development; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism; D006023:Glycoproteins / Q000235:genetics*; D049452:Green Fluorescent Proteins; D007814:Larva; D008164:Luminescent Proteins / Q000032:analysis / Q000235:genetics; D018613:Microscopy, Confocal; D009419:Nerve Tissue Proteins / Q000235:genetics*; D009420:Nervous System / Q000166:cytology* / Q000254:growth & development*; D011679:Pupa; D011993:Recombinant Fusion Proteins / Q000032:analysis / Q000096:biosynthesis; D012045:Regulatory Sequences, Nucleic Acid; D029701:Saccharomyces cerevisiae Proteins; D000254:Sodium-Potassium-Exchanging ATPase / Q000235:genetics; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism; D014162:Transfection
D004268:DNA-Binding Proteins; D029721:Drosophila Proteins; D005656:Fungal Proteins; C052130:GAL4 protein, S cerevisiae; D006023:Glycoproteins; D008164:Luminescent Proteins; D009419:Nerve Tissue Proteins; C094061:Nrv2 protein, Drosophila; D011993:Recombinant Fusion Proteins; D029701:Saccharomyces cerevisiae Proteins; D014157:Transcription Factors; D049452:Green Fluorescent Proteins; D000254:Sodium-Potassium-Exchanging ATPase