Genetic mechanisms specifying astrocyte functional diversity and their role in sleep

项目摘要 星形胶质细胞在进化上是保守的，在大脑中占相当大的比例，但我们的 对它们的身份和功能的了解远不如对神经元的全面。星形胶质细胞 被认为对调节神经元回路的形成和功能很重要，在 神经递质再摄取、离子缓冲、代谢支持和可塑性。来自哺乳动物和 果蝇也表明，星形胶质细胞在功能和形态上都是不同的。然而， 这种多样性的潜在遗传机制和功能重要性仍不清楚。这件事的重点是 GRANT是为了描述成年果蝇中央脑内星形胶质细胞的功能和分子多样性 以及它们在睡眠中的作用，这是一种保守的先天行为。在目标1中，我将系统地研究基因调控。 星形胶质细胞的异质性，特别是它们的区域规格和形态。我还将使用单细胞RNA 测序以确定星形胶质细胞内差异表达的基因并可视化时空模式 中央脑内星形胶质细胞的基因表达。这一目标的实现将为 果蝇星形胶质细胞特性和形态的综合分类。在目标2中，我将研究 用活体钙成像研究睡眠和觉醒过程中星形胶质细胞-神经元的局部相互作用 电生理学和行为分析。在R00阶段的目标3中，我将生成工具以使 靶向特定星形胶质细胞群体的交叉试剂，将广泛用于标记和 功能操纵。我将使用这些工具进一步剖析星形胶质细胞在调控中的不同角色 睡眠行为。由这一提议产生的研究将极大地扩展我们对星形胶质细胞生物学的知识， 通过1)识别负责星形胶质细胞功能和多样性的基因和分子，2)阐明它们的 与神经元回路的相互作用及其如何调节行为，以及3)针对和 操纵这种重要的细胞类型。重要的是，培训计划，包括课程作业，专业 开发，以及神经生物学、星形胶质细胞和遗传工具制造专家的指导，将准备 我未来的职业生涯是一名独立的学术神经学家。

Project Summary Astrocytes are evolutionarily conserved and constitute a substantial proportion of the cells in the brain, yet our understanding of their identities and functions is far less comprehensive than for neurons. Astrocytes are thought to be important for regulating the formation and function of neuronal circuits, playing multiple roles in neurotransmitter reuptake, ionic buffering, metabolic support, and plasticity. Evidence from both mammals and Drosophila has also indicated that astrocytes are diverse in both function and morphology. However, the underlying genetic mechanisms and the functional importance of this diversity remain unclear. The focus of this grant is to characterize the functional and molecular diversity of astrocytes in the adult Drosophila central brain and their role in sleep, a conserved innate behavior. In Aim 1, I will systematically investigate genes regulating astrocyte heterogeneity, specifically their regional specification and morphology. I will also use single cell RNA sequencing to identify differentially expressed genes within astrocytes and visualize the spatiotemporal pattern of gene expression in astrocytes in the central brain. Completion of this aim will lay the foundation for a comprehensive cataloging of astrocyte identity and morphology in Drosophila. In Aim 2, I will study the role of local astrocyte-neuron interactions in regulating sleep and arousal using in vivo calcium imaging, electrophysiology and behavioral assays. In Aim 3 during the R00 phase, I will generate tools to make stable intersectional reagents for targeting of specific astrocyte populations that will be broadly useful for labeling and functional manipulation. I will use these tools to further dissect the diverse roles of astrocytes in regulating sleep behavior. The research arising from this proposal will greatly expand our knowledge of astrocyte biology, by 1) identifying the genes and molecules responsible for astrocyte function and diversity, 2) elucidating their interactions with neuronal circuits and how they regulate behavior, and 3) development tools to target and manipulate this important cell type. Importantly, the training plan, including coursework, professional development, and mentorship from experts in neurobiology, astrocytes, and genetic tool-building, will prepare me for a future career as an independent academic neuroscientist.