Li Qiuling, Stavropoulos Nicholas
Department of Neuroscience and Physiology, Neuroscience Institute, New York University School of Medicine, New York 10016.
Department of Neuroscience and Physiology, Neuroscience Institute, New York University School of Medicine, New York 10016
G3 (Bethesda). 2016 Oct 13;6(10):3351-3359. doi: 10.1534/g3.116.034132.
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.
黑腹果蝇是一种用于剖析调节睡眠分子机制的强大模式生物,对果蝇的众多研究已经鉴定出影响睡眠-觉醒周期的基因。条件性遗传分析对于区分这些基因影响睡眠的机制至关重要:一些基因可能在发育过程中发挥作用,例如通过指导调节睡眠的神经回路的组装;其他基因可能在成年期调节睡眠;还有一些基因可能通过发育和成年机制来影响睡眠。在这里,我们评估了两种配体诱导表达系统,即Geneswitch和Q系统,用于果蝇睡眠的条件性和神经元特异性操纵。虽然如先前所示,成年特异性诱导神经元表达的Geneswitch转基因(elav-GS)与睡眠研究兼容,但elav-GS的发育诱导会强烈且非特异性地扰乱成年果蝇的睡眠。在低剂量的Geneswitch激动剂存在下以及在存在与睡眠无关的转基因(如UAS-CD8-GFP)时,elav-GS动物的睡眠就会发生改变。此外,发育性elav-GS诱导具有毒性并会减少产卵量,这表明神经元Geneswitch激活会产生多种不良影响。相比之下,当用于组成性、发育性或成年特异性神经元诱导时,Q系统的转基因和配体不会显著影响睡眠-觉醒周期。发育性elav-GS激活对睡眠的非特异性影响表明,此类操纵需要谨慎解读,并表明Q系统或其他策略可能更适合于果蝇睡眠及其他行为的条件性遗传分析。