The role of P2Y6 receptors in microglial calcium signaling: Investigations in the awake animal during the basal state and epilepsy development

P2Y6 受体在小胶质细胞钙信号传导中的作用：在清醒动物基础状态和癫痫发展过程中的研究

• 批准号: 10377815
• 负责人: ANTHONY DAVID UMPIERRE
• 金额: $ 3.39万
• 依托单位: MAYO CLINIC ROCHESTER
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-17 至 2021-09-16
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10377815
• 关键词: Adult; Animals; Astrocytes; Benchmarking; Brain; Calcium; Calcium Signaling; Cells; Center for Translational Science Activities; Clinic; Coupled; Development; Disease; Environment; Epilepsy; Epileptogenesis; Event; Functional disorder; GTP-Binding Proteins; Health; ITPR1 gene; Image; Immune; Infusion procedures; Injections; Injury; Investigation; Lasers; Longitudinal Studies; Mentors; Microglia; Microscopy; Modeling; Molecular; Mus; National Institute of Neurological Disorders and Stroke; Nerve Degeneration; Neuraxis; Neuroglia; Neuronal Injury; Neurons; Nucleotides; Pathway interactions; Phagocytosis; Pharmacology; Physiology; Play; Process; Purinoceptor; Research; Role; Seizures; Signal Transduction; Signaling Protein; Status Epilepticus; Structure; Surveys; Synapses; System; Testing; Time; Training; awake; brain parenchyma; calcium indicator; cell motility; cell type; clinical care; cytokine; designer receptors exclusively activated by designer drugs; extracellular; in vivo; kainate; knockout gene; nervous system disorder; neuronal cell body; purinoceptor P2Y6; receptor; two photon microscopy; two-photon

Project Summary/Abstract [This is the original document submitted in the F32 application. It is unchanged by this supplement.] Microglia are the resident immune cells of the central nervous system (CNS). In the CNS, calcium signaling is an essential secondary messenger system in astrocytes and neurons. However, in microglia, calcium signaling is not completely understood. In this proposal, we investigate microglial calcium signaling in health and disease. In our first aim, we explore whether microglial calcium signaling occurs in the awake mouse and whether that signaling is neuronal activity dependent using in vivo two-photon microscopy. Activity-dependent calcium signaling in microglia would suggest that this cell type could critically sense and respond to neuronal changes. The ability to sense neuronal activity may be particularly important during epilepsy development. In our second aim, we explore whether and how microglial calcium activity changes in a model of epilepsy development. In aim 2, we use in vivo, longitudinal two-photon imaging to study changes in calcium signaling during epileptogenesis. We hypothesize that the purinergic receptor, P2Y6, is essential for microglial calcium signaling in both health and disease. Our training plan utilizes selective pharmacology and gene knockout strategies to understand the mechanisms of microglial calcium signaling. Our proposed research is an important first step in understanding a critical secondary messenger system and its role in microglial-neuron interactions. Successful completion of these aims will advance our understanding of the epileptogenic process (NINDS Benchmark II for Epilepsy Research). Research will be performed at the Mayo Clinic, a renowned center for translational research and clinical care. Project mentors are experts in microglial physiology/pathophysiology (Dr. Long-Jun Wu), and mechanisms of epileptogenesis (Dr. Greg Worrell).

项目总结/摘要 [This是F32申请中提交的原始文件。这一补充没有改变）。 小胶质细胞是中枢神经系统（CNS）的常驻免疫细胞。在中枢神经系统中，钙信号是 星形胶质细胞和神经元中的一种重要的第二信使系统。然而，在小胶质细胞中，钙信号 还没有完全被理解。在这项提案中，我们研究了健康和疾病中的小胶质细胞钙信号。 在我们的第一个目标中，我们探索小胶质细胞钙信号是否发生在清醒的小鼠中， 使用体内双光子显微镜，信号传导取决于神经元活动。活动依赖性钙 小神经胶质细胞中的信号传导表明这种细胞类型可以关键地感知和响应神经元的变化。 感知神经元活动的能力在癫痫发展过程中可能特别重要。在我们的第二 目的是探索癫痫发展模型中小胶质细胞钙活性是否以及如何变化。在 目的2，我们使用体内纵向双光子成像来研究钙信号的变化， 癫痫发生我们假设嘌呤能受体P2 Y 6是小胶质细胞钙信号传导所必需的 无论是健康还是疾病。我们的训练计划利用选择性药理学和基因敲除策略， 了解小胶质细胞钙信号的机制。 我们提出的研究是理解关键的第二信使系统的重要的第一步， 它在小胶质细胞-神经元相互作用中的作用。成功地完成这些目标将促进我们的理解 NINDS Benchmark II for Epilepsy Research（NINDS癫痫研究基准II）研究将在 马约诊所，一个著名的转化研究和临床护理中心。项目导师是 小胶质细胞生理学/病理生理学（Dr. Long-Jun Wu）和癫痫发生机制（Dr. Greg Worrell）。