Identification of Biomarkers Reflecting Homeostatic Sleep Drive

两个相互作用的过程，生物钟和自我平衡的睡眠驱动，这反映了两个 唤醒经验的质量和数量，调节睡眠。生物钟已经被大量研究， 其背后的分子过程是很好理解的。另一方面，睡眠的自我调节能力很差， 我们对控制它的分子过程的了解充其量只是刚刚起步。的发现 基因周期，它编码分子钟的一个组成部分，并显示振荡表达 反映正常的生物钟功能，允许实时监测生物钟的分子机制 使用生物发光荧光素酶报告子（Per-Luc）。相比之下，睡眠驱力的最佳标志是 脑电图三角带功率，既不完全反映睡眠驱动，也不作为一个 睡眠平衡器的功能成分。因此，该项目的主要目标是使用一组特定的 定义理想稳态标志物的参数，以实际识别稳态睡眠驱力的标志物。 以前试图通过转录组学来鉴定在睡眠稳态中起作用的基因， 缺乏针对性基因表达的实质性变化通常可以在特定的细胞中观察到。 当全局分析显示没有的时候，电路。该提案利用了Allada实验室的专业知识， 基因表达谱的小，遗传定义的神经元群体，以确定转录，反映 果蝇大脑睡眠相关区域内稳态驱动的变化。此外，我还提出了几个新的 通过消除昼夜节律的影响和操纵来改善稳态基因检测的策略 在稳态驱动的恢复睡眠之前的清醒体验。候选人的成绩单将受到 二次筛选以消除在稳态驱动中不起功能作用的基因。最后我 建议开发新的工具，用于体内监测控制睡眠驱动的分子过程，以促进 未来对睡眠机制的研究。

Two interacting processes, the circadian clock and a homeostatic sleep drive, which reflects both the quality and quantity of waking experience, regulate sleep. The circadian clock has been heavily studied and the molecular process underlying it is well understood. The sleep homeostat, on the other hand, is poorly understood and our knowledge of the molecular processes governing it is incipient at best. Discovery of the gene period, which encodes an integral component of the molecular clock and displays oscillatory expression reflecting normal clock function, allowed for real-time monitoring of the clock's molecular mechanism in vivo using a bioluminescent luciferase reporter (per-luc). In contrast, the best marker of sleep drive is electroencephalogram delta band power, which neither completely reflects sleep drive nor serves as a functional component of the sleep homeostat. Thus, the primary goal of this project is to use a specific set of parameters defining an ideal homeostatic marker to actually identify markers of homeostatic sleep drive. Previous attempts to identify genes with a role in the sleep homeostat through transcriptomics have largely suffered from a lack of specificity. Substantial changes in gene expression can often be observed in specific circuits at times when global profiling reveals naught. This proposal leverages the Allada lab's expertise in gene expression profiling of small, genetically defined populations of neurons to identify transcripts that reflect changes in homeostatic drive in sleep relevant areas of the fly brain. In addition, I am proposing several novel strategies to improve detection of homeostatic genes by eliminating circadian influence and manipulating waking experience prior to homeostatically driven recovery sleep. Candidate transcripts will be subjected to secondary screening to eliminate genes that do not play a functional role in homeostatic drive. Finally, I propose to develop novel tools for in vivo monitoring of the molecular process governing sleep drive to facilitate future inquiry into the mechanisms underlying sleep.