Yoshii Taishi, Wülbeck Corinna, Sehadova Hana, Veleri Shobi, Bichler Dominik, Stanewsky Ralf, Helfrich-Förster Charlotte
Institute of Zoology, University of Regensburg, 93040 Regensburg, Germany.
J Neurosci. 2009 Feb 25;29(8):2597-610. doi: 10.1523/JNEUROSCI.5439-08.2009.
The neuropeptide pigment-dispersing factor (PDF) is a key transmitter in the circadian clock of Drosophila melanogaster. PDF is necessary for robust activity rhythms and is thought to couple the circadian oscillations of the clock neurons. However, little is known about the action of PDF on individual clock neurons. Here, we combined the period-luciferase reporter system with immunolabeling of clock proteins in wild-type and Pdf(01) mutants to dissect the effects of PDF on specific subgroups of clock neurons. Additionally, PDF levels were elevated to higher than normal levels using specific neural mutants, and a correlation analysis of locomotor activity and clock protein staining served to determine the periods of specific clock cells. We found that PDF has multiple effects on the clock neurons: In some groups of clock neurons, PDF was required for maintaining the oscillations of individual cells, and in others, PDF was required for synchronous cycling of the individual members. Other clock neurons cycled with high amplitude in absence of PDF, but PDF affected their intrinsic clock speed. Sometimes PDF shortened and sometimes PDF lengthened period. Our observations indicate that PDF is crucial for adjusting cycling amplitude, period, and phase of the different players in the circadian clock. Under natural conditions PDF may be required for adapting Drosophila's clock to varying photoperiods. Indeed, we show here that Pdf(01) mutants are not able to adapt their activity to long photoperiods in a wild-type manner.
神经肽色素分散因子(PDF)是黑腹果蝇昼夜节律钟的关键递质。PDF对于强健的活动节律是必需的,并且被认为能使时钟神经元的昼夜节律振荡相互耦合。然而,关于PDF对单个时钟神经元的作用却知之甚少。在此,我们将周期荧光素酶报告系统与野生型和Pdf(01)突变体中时钟蛋白的免疫标记相结合,以剖析PDF对时钟神经元特定亚群的影响。此外,利用特定的神经突变体将PDF水平提高到高于正常水平,并且对运动活动和时钟蛋白染色进行相关性分析,以确定特定时钟细胞的周期。我们发现PDF对时钟神经元有多种作用:在某些时钟神经元组中,PDF是维持单个细胞振荡所必需的,而在其他组中,PDF是单个成员同步循环所必需的。其他时钟神经元在没有PDF的情况下以高振幅循环,但PDF会影响它们的内在时钟速度。有时PDF会缩短周期,有时会延长周期。我们的观察结果表明,PDF对于调节昼夜节律钟中不同参与者的循环振幅、周期和相位至关重要。在自然条件下,PDF可能是使果蝇的生物钟适应不同光周期所必需的。事实上,我们在此表明,Pdf(01)突变体无法像野生型那样使其活动适应长光周期。