Investigation of Neuropeptide Signaling Mechanisms that Control Sleep

睡眠障碍越来越常见，并且与多种合并症和其他公共疾病有关。 健康后果。因此，提高我们对控制神经机制的理解至关重要 睡眠的时间和质量。调节果蝇等动物睡眠的关键信号分子 人类来自神经肽递质家族。这些分子的表达水平稀疏并且 对行为（包括睡眠）的选择性影响，使他们成为发展专注力的最佳人选 副作用最小的药物治疗。然而，这些分子单独作用的机制 并协同调节大脑中的靶细胞，因此我们对行为知之甚少。此次续订 应用程序将利用强大的遗传学和相对简单的睡眠网络组织 果蝇（Drosophila melanogaster），研究神经肽如何在分子、细胞和 调节睡眠的行为水平。在目标 1 中，我们将利用特定神经元子集的光遗传学激活 表达促进睡眠的递质短神经肽 F (sNPF)，以确定其特定的 睡眠调节中的作用。在目标 2 中，我们将共同激活促进睡眠的 sNPF 神经元和促进觉醒的神经元 神经元来确定它们在睡眠调节中的层次结构。在目标 3 中，我们将利用活体荧光 成像来检验 sNPF 作为白细胞激肽神经元的抑制性输出信号的假设 睡觉。结合起来，拟议的工作将使我们能够确定 sNPF 神经元在睡眠中如何发挥作用 大脑中的调节网络，扩大了我们对控制睡眠和睡眠机制的理解 分子和电路层面的觉醒。反过来，这些知识将为更多的设计提供基础 有效治疗人类睡眠异常。此次R15 AREA更新提案将直接涉及 本科生参与研究的各个方面，包括设计和开展涉及的研究 基因操作、分子和细胞神经生物学以及行为分析技术。学生将 还积极传播通过研究获得的科学知识，以 会议演讲和出版的手稿。他们的经验将为未来提供形成性培训 生物医学领域的职业。

Sleep disturbances are increasingly common and are associated with a variety of comorbidities and other public health consequences. It is therefore critical to improve our understanding of the neural mechanisms that control the timing and quality of sleep. Key signaling molecules that regulate sleep in animals ranging from flies to humans come from the family of neuropeptide transmitters. These molecules have sparse expression levels and selective effects on behavior, including sleep, making them prime candidates for the development of focused drug treatments with minimal side effects. However, the mechanisms by which these molecules act individually and in concert to regulate target cells in the brain and thus behavior are poorly understood. This renewal application will take advantage of the powerful genetics and relatively simple sleep network organization of the fruit fly, Drosophila melanogaster, to address how neuropeptides function at the molecular, cellular, and behavioral levels to regulate sleep. In Aim 1, we will utilize optogenetic activation of specific subsets of neurons expressing the sleep-promoting transmitter short neuropeptide F (sNPF) in order to determine their particular roles in sleep regulation. In Aim 2, we will coactivate sleep-promoting sNPF neurons and wake-promoting neurons to determine their hierarchical organization in sleep regulation. In Aim 3, we will utilize live fluorescent imaging to test the hypothesis that sNPF acts as an inhibitory output signal onto leucokinin neurons that regulate sleep. In combination, the proposed work will allow us to establish how sNPF neurons function within sleep regulatory networks in the brain, expanding our understanding of the mechanisms that control sleep and wakefulness at the molecular and circuit levels. In turn, this knowledge will provide a basis for the design of more effective treatments of human sleep abnormalities. This R15 AREA renewal proposal will directly involve undergraduate students in all aspects of the research, including designing and carrying out studies involving techniques of genetic manipulation, molecular and cellular neurobiology, and behavioral analysis. Students will also take an active role in disseminating the scientific knowledge acquired through their research, in the form of conference presentations and published manuscripts. Their experiences will provide formative training for future careers in biomedical fields.