Analysis of activity-dependent interactions between microglia and synapses

在我读K99的第一年，我收到了一份工作，并接受了一份助理教授的职位。 马萨诸塞大学医学院神经生物学系。因此，我正在申请 过渡到Roo阶段，在那里我将研究驱动活动依赖的小胶质细胞特有的机制 发育中的大脑中的突触重构。最终目标是将这些机制应用于 神经发育和神经精神障碍。 不成熟的突触形成了一个粗略的接线图，必须在发育过程中重新塑造，以实现 成熟神经系统的精确连接特征。虽然很明显，神经活动推动 突触重塑，其潜在机制尚不完全清楚。我们最近发现， 小胶质细胞以一种活性依赖的方式吞噬突触成分，参与突触重构。 然而，尚不清楚小胶质细胞是否吞噬了完整的突触，或者这是否是非细胞自主性突触。 事件。此外，已知小胶质细胞改变它们的运动性和与突触的相互作用作为反应 神经活性，但这些反应的潜在分子机制和功能后果 都是未知的。因此，这项提议的目标是：1)AIMI：检验小胶质细胞是 主动感知、相互作用和吞噬完整突触以响应神经变化 活动。该实验室在小胶质细胞成像方面的专业知识是解决这一问题的独特之处。 双光子活体成像，这是在K99阶段获得的一项技能。2)目标2：解剖分子 小胶质细胞对神经活动变化作出反应的潜在机制。一些候选的小胶质细胞特异的 分子和细胞因子已经被确认和验证，包括IL-12。因此，依赖于活动 小胶质细胞的运动性和与突触的相互作用将在基因缺陷的小鼠中进行评估 利息。3)目标3：通过以下方式确定活性依赖的小胶质细胞反应的功能意义 检验这些反应调节突触回路结构和突触发育的假设 先前发现和验证基因缺陷的小鼠的功能。

During the first year of my K99, I received an offer and accepted a positon as an Assistant Professor in the Neurobiology Department at the University of Massachusetts Medical School. As a result, I am applying for transition to the ROO phase where I will be investigating microglia-specific mechanisms driving activity-dependent synaptic remodeling in the developing brain. The ultimate goal is to apply these mechanisms to neurodevelopmental and neuropsychiatric disorders. Immature synapses form a crude wiring diagram that must remodel during development to achieve the precise connectivity characteristic of the mature nervous system. While It is clear that neural activity drives synaptic remodeling, the underlying mechanisms are not fully understood. We recently identified that microglia participate in synaptic remodeling by engulfing synaptic elements in an activity-dependent manner. However, it is unknown whether microglia engulf intact synapses or whether this is a non-cell autonomous event. In addition, it is known that microglia change their motility and interactions with synapse in response to neural activity but the underlying molecular mechanisms and functional consequences of these responses are unknown. As a result the goals of this proposal are to: 1) Aimi: Test the hypothesis that microglia are actively sensing, interacting with, and phagocytosing intact synapses in response to changes in neural activity. The laboratory is uniquely positioned to address this question with expertise in imaging microglia by 2-photon In vivo live Imaging, a skill acquired during the K99 phase. 2) Aim 2: Dissect the molecular mechanisms underlying microglia responses to changes in neural activity. A number of candidate microglia-specific molecules and cytokines have been identified and validated, including IL-12. Thus, activity-dependent microglia motility and interactions with synapses will be assessed In mice deficient in genes of interest. 3) Aim 3: Determine the functional significance of activity-dependent microglial responses by testing the hypothesis that these responses regulate the development of synaptic circuit structure and function in mice with deficiencies in genes previously identified and validated.