Circadian Regulation of the PER2::LUC Bioluminescence Rhythm in the Mouse RPE

描述（由申请人提供）：视网膜色素上皮（RPE）在维持光感受器的健康和功能方面起着重要作用。先前的研究表明，RPE还参与了光盘脱落的调节，这一过程对光感受器的健康至关重要。这一过程已被证明是受昼夜节律控制的，尽管控制昼夜节律的机制仍然知之甚少。我们的实验室已经开发出一种新的制剂，可以使用PER2::LUC小鼠轻松监测RPE的昼夜节律。我们的初步数据表明，从培养的RPE中可以记录到PER2::LUC生物发光的昼夜节律，这种节律的阶段与视网膜中观察到的阶段不同。光确实使RPE的昼夜节律发生相移，而SCN病变不影响这种节律的相位。有趣的是，我们发现多巴胺的使用可以改变RPE的昼夜节律。在这项探索性应用中，我们建议研究多巴胺和多巴胺受体在RPE昼夜节律调节中的作用以及RPE转录组的昼夜节律调节。

DESCRIPTION (provided by applicant): The retinal pigment epithelium (RPE) plays an important role in the maintenance of photoreceptors health and functioning. Previous studies have shown that RPE is also involved in the regulation of the disc shedding, a process that is that is vital for the health of the photoreceptors. This process has been shown to be under circadian control, albeit the mechanisms controlling this circadian rhythm are still poorly understood. Our laboratory has developed a novel preparation in which circadian rhythms in RPE can be easily monitored using the PER2::LUC mouse. Our preliminary data indicate that a circadian rhythm in PER2::LUC bioluminescence can be recorded from cultured RPE and the phase of such a rhythm is different from the phase observed in the retina. Light does phase-shift the circadian rhythm in the RPE, and SCN lesion does not affect the phase of this rhythm. Interestingly, we discovered that Dopamine application can shift the circadian rhythm in the RPE. In this exploratory application, we propose to investigate the role of Dopamine and Dopamine receptors in the regulation of the circadian rhythms in the RPE and the circadian regulation of the RPE transcriptome. PUBLIC HEALTH RELEVANCE: The retinal pigment epithelium (RPE) plays an important role in the maintenance of photoreceptors health and functioning. Previous studies have shown that RPE is also involved in the regulation of the disc shedding, a process that is that is vital for the health of the photoreceptors. This process has been shown to be under circadian control, albeit the mechanisms controlling this circadian rhythm are still poorly understood. Modern life style has tremendously changed the time at which we expose ourselves to light; hence, it is important to understand how the circadian clock controls the rhythmic event in this organ. Indeed, the national plan for the eye and vision research of the National Eyes Institute states as one of its primary program objective the understanding of the mechanisms controlling the circadian shedding of photoreceptor outer segments and their phagocytosis by the RPE. In this grant application, we propose first to determine the role of DA in the entrainment of PER2::LUC circadian rhythm in the RPE and then to identify genes showing a circadian pattern of expression in RPE. This effort could prove a valuable tool to discovering genes and molecular pathways regulating rhythmic event in the RPE.