title: Evaluation of Ligand-Inducible Expression Systems for Conditional Neuronal Manipulations of Sleep in Drosophila.
abstract: Drosophila melanogaster is a powerful model organism for dissecting the molecular mechanisms that regulate sleep, and numerous studies in the fly have identified genes that impact sleep-wake cycles. Conditional genetic analysis is essential to distinguish the mechanisms by which these genes impact sleep: some genes might exert their effects developmentally, for instance by directing the assembly of neuronal circuits that regulate sleep; other genes may regulate sleep in adulthood; and yet other genes might influence sleep by both developmental and adult mechanisms. Here we have assessed two ligand-inducible expression systems, Geneswitch and the Q-system, for conditional and neuronally restricted manipulations of sleep in Drosophila While adult-specific induction of a neuronally expressed Geneswitch transgene (elav-GS) is compatible with studies of sleep as shown previously, developmental induction of elav-GS strongly and nonspecifically perturbs sleep in adults. The alterations of sleep in elav-GS animals occur at low doses of Geneswitch agonist and in the presence of transgenes unrelated to sleep, such as UAS-CD8-GFP Furthermore, developmental elav-GS induction is toxic and reduces brood size, indicating multiple adverse effects of neuronal Geneswitch activation. In contrast, the transgenes and ligand of the Q-system do not significantly impact sleep-wake cycles when used for constitutive, developmental, or adult-specific neuronal induction. The nonspecific effects of developmental elav-GS activation on sleep indicate that such manipulations require cautious interpretation, and suggest that the Q-system or other strategies may be more suitable for conditional genetic analysis of sleep and other behaviors in Drosophila.
authors: Department of Neuroscience and Physiology, Neuroscience Institute, New York University School of Medicine, New York 10016.
authors: Qiuling
authors: 0000-0002-6101-8779
authors: Q
authors: Li
authors: Department of Neuroscience and Physiology, Neuroscience Institute, New York University School of Medicine, New York 10016 nicholas.stavropoulos@nyumc.org.
authors: Nicholas
authors: 0000-0001-5915-2760
authors: N
authors: Stavropoulos
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D001921:Brain / Q000378:metabolism; D004330:Drosophila / Q000502:physiology*; D004331:Drosophila melanogaster / Q000235:genetics* / Q000378:metabolism; D005260:Female; D015870:Gene Expression; D005786:Gene Expression Regulation / Q000187:drug effects*; D023061:Gene Order; D005822:Genetic Vectors / Q000235:genetics; D005838:Genotype; D008024:Ligands; D008297:Male; D015735:Mifepristone / Q000494:pharmacology; D009928:Organ Specificity / Q000235:genetics; D012890:Sleep / Q000235:genetics*; D019076:Transgenes
chemical_list: D008024:Ligands; D015735:Mifepristone
keywords: Cul3; FlyBook; Geneswitch; Q-system; RU486; insomniac