Neuroscience Training Program, University of Wisconsin-Madison, Madison, WI 53706, USA.
PLoS One. 2012;7(10):e45130. doi: 10.1371/journal.pone.0045130. Epub 2012 Oct 15.
cAMP response element-binding protein (CREB) and nuclear factor kappa-B (NF-κB) are two ubiquitous transcription factors involved in a wide number of cellular processes, including the circadian system. Many previous studies on these factors use cellular assays that provide limited information on circadian activity or anatomical specificity. The ability to study transcription factors in defined tissue within intact animals will help to bridge the gap between cellular and in vivo data. We have used the GAL4-UAS and FLP-FRT systems to gain spatial control over reporter gene expression. Using a luciferase-based reporter, we show in vivo that Drosophila dCREB2- and NF-κB-mediated transcription oscillates in neuronal cells, glia, and in the mushroom body, a higher-order brain center in flies. This oscillation is under circadian control, cycling with a 24-hour rhythm, under both light-dark and dark-dark conditions. In light-light conditions, dCREB2 and NF-κB reporter flies exhibit a suppression of rhythmic activity. Furthermore, neuronal cycling of dCREB2 and NF-κB activity are modulated in period mutant flies, indicating these oscillations are controlled through the central clock. This study shows for the first time region-specific circadian oscillation of dCREB2/NF-κB activity in the Drosophila nervous system.
环腺苷酸反应元件结合蛋白 (CREB) 和核因子 kappa-B (NF-κB) 是两种普遍存在的转录因子,参与广泛的细胞过程,包括生物钟系统。许多关于这些因子的先前研究使用细胞测定法,这些方法提供了关于生物钟活动或解剖特异性的有限信息。在完整动物中研究特定组织中的转录因子的能力将有助于弥合细胞和体内数据之间的差距。我们使用 GAL4-UAS 和 FLP-FRT 系统来获得对报告基因表达的空间控制。使用基于荧光素酶的报告基因,我们在体内表明,果蝇 dCREB2 和 NF-κB 介导的转录在神经元细胞、神经胶质细胞和蘑菇体中(果蝇的高级脑中心)呈振荡状态。这种振荡受生物钟控制,在光-暗和暗-暗条件下以 24 小时的节律循环。在光-光条件下,dCREB2 和 NF-κB 报告果蝇表现出节律活动的抑制。此外,在周期突变体果蝇中,神经元的 dCREB2 和 NF-κB 活性循环被调节,表明这些振荡是通过中央时钟控制的。这项研究首次表明,果蝇神经系统中 dCREB2/NF-κB 活性具有区域特异性的生物钟振荡。