Astrocyte modulation of sleep

星形胶质细胞对睡眠的调节

• 批准号: 9977360
• 负责人: F Rob JACKSON
• 金额: $ 45.38万
• 依托单位: TUFTS UNIVERSITY BOSTON
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-04-01 至 2022-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9977360
• 关键词: Adult; Affect; Affinity Chromatography; Arousal; Astrocytes; Behavior; Behavioral Genetics; Binding; Brain; Brain region; Cells; Communication; Drosophila genus; Expression Profiling; Future; Gene Expression; Gene Expression Profile; Genes; Genetic study; Goals; Homeostasis; Immunoglobulin Domain; Immunoglobulins; Individual; Invertebrates; Knowledge; Letters; Life; Mammals; Mediating; Membrane; Methods; Modeling; Molecular; Monitor; Morphology; Neuroglia; Neurons; Orthologous Gene; Pathway interactions; Pattern; Peptides; Periodicity; Population; Process; Proteins; Publications; Publishing; Receptor Signaling; Reporter; Research; Ribosomes; Signal Pathway; Signal Transduction; Sleep; Synapses; System; Technology; Temperature; Tertiary Protein Structure; Testing; Translating; Vesicle; Wakefulness; autocrine; base; cell type; cellular targeting; circadian; circadian regulation; experience; fly; genetic manipulation; genome-wide; in vivo; insight; interest; knock-down; nervous system disorder; neural circuit; neuronal circuitry; neuronal excitability; novel; receptor; response; sensor; sleep regulation; trafficking; transcriptome sequencing; vesicular release; voltage

Endogenous rhythmic behaviors are evolutionarily conserved and essential for life. Both in mammalian and invertebrate models, there are known requirements for glial cells, in addition to neuronal circuits, in the regulation of circadian behavior and sleep 1-4. Using the Drosophila model, our lab was the first to demonstrate that astrocytes were required, in vivo, for normal circadian behavior 5, 6, and we show in this proposal that astrocytes also modulate sleep. In preliminary studies for this application, we have demonstrated that activation of fly astrocytes, by increasing intracellular Ca2+, promotes sleep whereas inhibition of astrocyte vesicle trafficking and release leads to decreased sleep. In related studies (now published), we have shown that an astrocyte-enriched, secreted protein known as NKT promotes sleep 7. Based on these findings, we hypothesize that neuronal sleep-regulating circuits are subject to astrocyte modulation. Aim 1a will test this hypothesis by simultaneously activating or inhibiting astrocytes and recording the excitability of known sleep- regulating neuronal populations using genetically encoded voltage sensors. This will identify neuronal groups that show altered excitability with activation or inhibition of astrocytes. In complementary studies of Aim 1b, we will express membrane-tethered versions of NKT in all known sleep-regulating neuronal groups to identify those subject to modulation by the glial protein. NKT is a secreted astrocyte protein, but this subaim will utilize neuronal expression of membrane-tethered NKTs to identify its cellular targets. Together, Aims 1a and 1b will identify sleep-regulating neurons that are subject to astrocyte modulation. Aim 2 will use Translating Ribosome Affinity Purification (TRAP) methods combined with genome-wide RNA-seq to test the hypothesis that the gene expression profile of sleep-regulating neurons is altered by astrocyte modulation. The studies of this aim will focus on identification of neuronal factors (receptors, intracellular signaling components) with vertebrate orthologs that may mediate the astrocyte modulation of sleep. Preliminary results documenting feasibility for the aims are included in the proposal. Together, the two aims will provide valuable information about neuronal targets of astrocyte modulation and identify neuronal molecular pathways that respond to astrocyte signals. Such pathways may be important for glia-to-neuron communication or more directly the regulation of sleep.

内源性节律行为在进化上是保守的，对生命至关重要。在哺乳动物和 在无脊椎动物模型中，除了神经元回路之外， 昼夜节律行为和睡眠的调节1-4.利用果蝇模型，我们的实验室是第一个证明 星形胶质细胞是体内正常昼夜节律行为所必需的5，6，我们在这一提议中表明， 星形胶质细胞也调节睡眠。在这项应用的初步研究中，我们已经证明， 通过增加细胞内Ca 2+激活果蝇星形胶质细胞，促进睡眠，而抑制星形胶质细胞 囊泡的运输和释放导致睡眠减少。在相关研究（现已出版）中，我们已经表明， 一种富含星形胶质细胞的分泌蛋白NKT促进睡眠。基于这些发现，我们 假设神经元睡眠调节回路受星形胶质细胞调节。Aim 1a将测试这一点 通过同时激活或抑制星形胶质细胞并记录已知睡眠的兴奋性， 使用遗传编码的电压传感器调节神经元群体。这将识别神经元组 其显示出随着星形胶质细胞的激活或抑制而改变的兴奋性。在目标1b的补充研究中，我们 将在所有已知的睡眠调节神经元群中表达NKT的膜系版本，以确定 那些受神经胶质蛋白调节的细胞。NKT是一种分泌型星形胶质细胞蛋白，但该子目标将利用 神经元表达的膜系NKT，以确定其细胞靶点。目标1a和1b将共同 识别受星形胶质细胞调节的睡眠调节神经元。目标2将使用翻译核糖体 亲和纯化（TRAP）方法与全基因组RNA-seq相结合，以检验基因 睡眠调节神经元的表达谱通过星形胶质细胞调节而改变。对这一目标的研究将 专注于脊椎动物神经元因子（受体，细胞内信号成分）的鉴定 可能介导睡眠的星形胶质细胞调节的直系同源物。初步结果记录了 这些目标已包括在提案中。总之，这两个目标将提供有关神经元的有价值的信息。 星形胶质细胞调节的靶点，并识别响应星形胶质细胞信号的神经元分子通路。 这些通路可能对神经胶质细胞与神经元的通讯或更直接的睡眠调节很重要。