Analysis of activity-dependent interactions between microglia and synapses

小胶质细胞和突触之间活动依赖性相互作用的分析

• 批准号: 8618105
• 负责人: Dorothy Patricia Schafer
• 金额: $ 9万
• 依托单位: BOSTON CHILDREN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-01-01 至 2014-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8618105
• 关键词: Address; Affect; Autistic Disorder; Automobile Driving; Award; Boston; Brain; Candidate Disease Gene; Cells; Characteristics; Data; Development; Developmental Process; Disease; Elements; Event; Functional disorder; Funding; Future; Gene Delivery; Gene Expression; Genes; Genetic; Goals; Image; Immediate-Early Genes; Immune; Immunohistochemistry; In Vitro; Indium; Interleukin 12 Receptor Beta; Interleukin-12; Knockout Mice; Label; Laboratories; Learning; Life; Link; Maintenance; Measures; Mediating; Mental disorders; Mentors; Microglia; Modeling; Molecular; Mus; Nervous system structure; Neurodevelopmental Disorder; Neuroglia; Neurons; Pathology; Pathway interactions; Pediatric Hospitals; Phagocytes; Phagocytosis; Phase; Physiology; Play; Presynaptic Terminals; Process; Property; Proteins; Proteomics; Research Personnel; Resolution; Role; Schizophrenia; Sensory; Shapes; Signal Transduction; Structure; Study models; Synapses; System; Techniques; Testing; Time; Training; Translating; Viral; Virus; Visual; Visual system structure; Work; area striata; career; career development; cell type; density; design; experience; immune function; in vivo; in vivo imaging; interest; interleukin-12 receptor; medical schools; neural circuit; postnatal; postsynaptic; public health relevance; receptor; relating to nervous system; research study; response; synaptogenesis; time use; two-photon

7. Project Summary/Abstract To achieve the exquisite precision characteristic of synaptic circuits in the mature nervous system, immature synapses form a crude wiring diagram that must remodel during postnatal development. While it is clear that neural activity drives developmental synaptic remodeling, the underlying mechanisms are not fully understood. This K99/R00 proposal will support my career development as I investigate underlying cellular and molecular mechanisms driving activity-dependent synaptic remodeling in the developing brain. I have firmly established that microglia, the resident CNS immune cells, phagocytose synaptic elements in the postnatal brain in response to changes in neural activity. These data raise the intriguing possibility that microglia are a cellular mechanism driving activity-dependent synaptic remodeling. However, it is unknown whether microglia actively phagocytose intact synapses or passively phagocytose synaptic remnants. In addition, the molecular mechanisms underlying activity-dependent microglia-synapse interactions are unknown. These questions will be addressed during the mentored phase of the award at Boston Children's Hospital and Harvard Medical School under the guidance of Dr. Beth Stevens and Dr. Michael Greenberg. Aim 1 is designed to test the hypothesis that microglia are actively phagocytosing intact synapses in response to changes in neural activity. In doing so, I will learn viral-mediated gene delivery to fluorescently label specific neural circuits and 2-photon in vivo live imaging to analyze microglia-synapse interactions in real time. Aim 2 is designed to investigate molecular mechanisms underlying these interactions. After genetic and proteomic screens, I have identified a candidate, interleukin 12 (IL-12). I will test the hypothesis that IL-12 regulates activity-dependent microglia-synapse interactions by in vivo imaging in IL-12 and IL-12 receptor KO mice. Specific Aim 3 (to be completed in my own laboratory) will determine the functional significance of activity- dependent microglia-synapse interactions by testing the hypothesis that these interactions regulate the functional development of synaptic circuits. I will genetically ablate microglia during a specific window of postnatal development and assess synapse structure and physiology. To manipulate microglia function more specifically, I will also test the role of the IL-12 pathway. Furthermore, as an alternative and/or future direction, my genetic and proteomic screens have identified several other putative molecular pathways that I will test. Given that aberrant synaptic circuits and microglial dysfunction have now been linked with several neurodevelopmental and psychiatric disorders, this proposal will have broad implications. My long term career goal as an independent investigator is to translate my findings to understand how microglia and other glial cell types may contribute to synaptic circuit abnormalities associated with disorders such as autism and schizophrenia. This award will help me to complete my training and provide an opportunity to learn techniques and obtain data for a successful R01 application to fund my future work.

7。项目摘要/摘要 为了实现成熟神经系统中突触电路的精确精确特征， 未成熟的突触形成了一个粗线图，该图在产后发育过程中必须重塑。虽然是 清楚的是，神经活动驱动发育突触重塑，基本机制并非完全 理解。当我调查基本的蜂窝细胞和 分子机制在发育中的大脑中驱动活性依赖性突触重塑。 我已经牢固地确定了小胶质细胞，常驻CNS免疫细胞，吞噬诱导的突触元素 产后大脑响应神经活动的变化。这些数据引起了有趣的可能性 小胶质细胞是驱动活动依赖性突触重塑的细胞机制。但是，这是未知的 小胶质细胞是主动吞噬诱导的完整突触还是被动吞噬吞噬突触残留物。在 此外，依赖活性依赖性小胶质细胞 - 共生相互作用的分子机制是 未知。这些问题将在波士顿儿童奖的指导阶段中解决 在贝丝·史蒂文斯（Beth Stevens）博士和迈克尔·格林伯格（Michael Greenberg）博士的指导下，医院和哈佛医学院。目的 1旨在检验小胶质细胞的假设是积极地吞噬完整的突触。 神经活动的变化。在此过程中，我将学习病毒介导的基因输送到荧光标记特定的标签 神经回路和2光子体内活体现场成像，以实时分析小胶质细胞的相互作用。 AIM 2是 旨在研究这些相互作用的分子机制。遗传和蛋白质组学后 屏幕，我已经确定了一个候选人，介绍了白介素12（IL-12）。我将测试IL-12调节的假设 IL-12和IL-12受体KO小鼠中体内成像的活性依赖性小胶质细胞间隔相互作用。 具体目标3（要在我自己的实验室完成）将确定活动的功能意义 - 通过测试这些相互作用调节这些相互作用的假设，依赖性小胶质细胞 - 共生相互作用 突触电路的功能发展。我将在特定窗口中遗传浸润小胶质细胞 产后发展和评估突触结构和生理学。操纵小胶质细胞功能更多 具体而言，我还将测试IL-12途径的作用。此外，作为替代方案和/或未来的方向， 我的遗传和蛋白质组学筛选已经确定了我将测试的其他几种推定的分子途径。 鉴于异常突触电路和小胶质细胞功能障碍现已与几个 神经发育和精神疾病，该提案将具有广泛的影响。我的长期职业 作为独立研究者的目标是翻译我的发现，以了解小胶质细胞和其他神经胶质细胞如何 类型可能导致突触电路异常与自闭症和诸如自闭症等疾病有关的异常 精神分裂症。该奖项将帮助我完成培训，并提供一个学习技术的机会 并获取成功的R01应用程序的数据，以资助我未来的工作。