MECHANISMS OF CIRCADIAN CLOCK OUTPUT

描述（由申请人提供）： 我们感兴趣的是昼夜节律起搏神经元组织日常运动行为的信号机制。近年来，在确定细胞自主发条机制的分子基础方面取得了巨大进展。这一定义允许识别大脑中的主要起搏器时钟神经元，并反过来提供了重新审视有关行为神经基础的基本问题的机会。在上一个资助周期中，我们确定了PDF的受体，PDF是果蝇昼夜神经回路中的主要递质。这一发现为目前的赠款申请奠定了基础。我们现在建议通过几种独立的方法来描述PDF受体的表达。这些信息对于帮助解释构成日常运动节律基础的PDF信号至关重要。 其次，我们将建立一个体外细胞培养模型，探讨PDF如何通过调节昼夜节律的分子振荡器机制来激活起搏神经元。有体内证据表明，PDF延迟PERIOD蛋白进入细胞核，这构成了一个有待检验的明确假设。在过去的一年中，我们已经采用了一种新的基因FRET报告测量cAMP水平在体内实时。因此，我们的第三个目标是使用这种方法来研究活脑中的PDF信号动力学。最后，我们将扩大我们的研究重点超越PDF追求的候选人资格的几个额外的神经递质/神经肽的果蝇昼夜神经回路的操作的潜在贡献。 昼夜节律起搏器机制的正常运作对于身体和认知功能的适当日常协调至关重要。当身体的计时机制出错时，如季节性适应性障碍或轮班工作时间表的结果，可能会导致临床并发症。我们所从事的工作将直接解决帮助同步身体神经元时钟的基本机制。我们帮助建立的原则将有助于开发可以逆转这些时间生物学紊乱的疗法。我们研究了昼夜节律起搏神经元组织日常运动行为的信号机制。我们面临着许多挑战，身体的正常计时功能，包括旅行相关的时差，轮班工作时间表，和季节性疾病。 我们工作的主要焦点是起搏神经元的递质信号，以帮助更多地了解可能有助于逆转这种时间相关疾病的过程。

DESCRIPTION (provided by applicant): We are interested in signaling mechanisms used by circadian pacemaker neurons to organize daily locomotor behavior. There has been tremendous progress in recent years to define the molecular basis of the cell autonomous clockwork mechanism. That definition has permitted the identification of the primary pacemaker clock neurons within the brain, and in turn presented the opportunity to re-examine fundamental issues concerning the neural basis of behavior. In the previous grant cycle, we identified the receptor for PDF, which is a primary transmitter in the Drosophila circadian neural circuit. That finding establishes a basis for the current grant application. We now propose to describe PDF receptor expression by several independent means. This information will be critical to help interpret PDF signaling that underlies daily locomotor rhythms. Second we will establish an in vitro cell culture model to explore how PDF synchronizes pacemaker neurons by regulating the circadian molecular oscillator mechanism. There is in vivo evidence that PDF delays the entry of PERIOD protein into the nucleus and that frames an explicit hypothesis to be tested. In the past year, we have adapted a novel genetic FRET reporter to measure cAMP levels in vivo real-time. Thus our third aims it to use this method to study PDF signaling dynamics in the living brain. Finally, we will expand our research focus beyond PDF by pursuing the candidacy of several additional neurotransmitters/ neuropeptides for their potential contributions to the operations of the Drosophila circadian neural circuit. The normal functioning of the circadian pacemaker mechanism is essential for proper daily coordination of body and cognitive functions. When the body's timekeeping mechanisms go awry, as in seasonal adaptive disorders or as a consequence of shift work schedules, clinical complications can result. The work we undertake will directly address the fundamental mechanisms that help synchronize the body's clockwork of neurons. The principles we help establish will be useful to develop therapies that can reverse these chronobiological disorders.We study signaling mechanisms used by circadian pacemaker neurons to organize daily locomotor behavior. We face numerous challenges to the body's normal timekeeping functions, including travel-related jet-lag, shift work schedules, and seasonal disorders. The major focus for our work concerns transmitter signaling by pacemaker neurons to help learn more about the processes that could help reverse such time-related disorders.