Targeted gene expression in Drosophila dopaminergic cells using regulatory sequences from tyrosine hydroxylase.
Dopamine (DA) is the only catecholaminergic neurotransmitter in the fruit fly Drosophila melanogaster. Dopaminergic neurons have been identified in the larval and adult central nervous system (CNS) in Drosophila and other insects, but no specific genetic tool was available to study their development, function, and degeneration in vivo. In Drosophila as in vertebrates, the rate-limiting step in DA biosynthesis is catalyzed by the enzyme tyrosine hydroxylase (TH). The Drosophila TH gene (DTH) is specifically expressed in all dopaminergic cells and the corresponding mutant, pale (ple), is embryonic lethal. We have performed ple rescue experiments with modified DTH transgenes. Our results indicate that partially redundant regulatory elements located in DTH introns are required for proper expression of this gene in the CNS. Based on this study, we generated a GAL4 driver transgene, TH-GAL4, containing regulatory sequences from the DTH 5' flanking and downstream coding regions. TH-GAL4 specifically expresses in dopaminergic cells in embryos, larval CNS, and adult brain when introduced into the Drosophila genome. As a first application of this driver, we observed that in vivo inhibition of DA release induces a striking hyperexcitability behavior in adult flies. We propose that TH-GAL4 will be useful for studies of the role of DA in behavior and disease models in Drosophila.
Friggi-Grelin
Florence
F
Laboratoire de Génétique et Physiologie du Développement, CNRS-Université de la Méditerranée, Developmental Biology Institute of Marseille, Campus de Luminy, Case 907, F-13288 Marseille Cedex 9, France.
Coulom
Hélène
H
Meller
Margaret
M
Gomez
Delphine
D
Hirsh
Jay
J
Birman
Serge
S
D000818:Animals; D030801:Animals, Genetically Modified; D002490:Central Nervous System / Q000378:metabolism / Q000502:physiology; D004268:DNA-Binding Proteins; D004298:Dopamine / Q000378:metabolism*; D004331:Drosophila melanogaster / Q000201:enzymology / Q000235:genetics; D018507:Gene Expression Regulation, Developmental; D015971:Gene Expression Regulation, Enzymologic; D017344:Genes, Insect; D005809:Genes, Regulator / Q000235:genetics / Q000502:physiology*; D007150:Immunohistochemistry; D017403:In Situ Hybridization; D007438:Introns; D007814:Larva / Q000235:genetics / Q000254:growth & development; D008666:Metalloendopeptidases / Q000235:genetics; D009154:Mutation; D013216:Reflex, Startle / Q000235:genetics / Q000502:physiology; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics; D013744:Tetanus Toxin / Q000235:genetics; D014157:Transcription Factors / Q000235:genetics; D014446:Tyrosine 3-Monooxygenase / Q000235:genetics / Q000378:metabolism*
D004268:DNA-Binding Proteins; C052130:GAL4 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; D013744:Tetanus Toxin; D014157:Transcription Factors; D014446:Tyrosine 3-Monooxygenase; D008666:Metalloendopeptidases; C093556:zinc-endopeptidase, tetanus neurotoxin; D004298:Dopamine