Novel approaches to study microglia physiology and pathology in the intact brain

描述（由申请人提供） 摘要：小胶质细胞是大多数（如果不是所有）中枢神经系统（CNS）疾病的关键参与者，包括神经性疼痛，Rett综合征，肌萎缩侧索硬化症和Nasu-Hakola病。最近的证据表明，这些细胞也参与调节神经发生，电路重塑和突触可塑性在正常的出生后发育，影响他们可能会继续发挥在成年大脑。尽管这些细胞在正常的生理和病理中起着重要的作用，但我们对这些细胞的信号传导，它如何受到局部环境和疾病的影响，以及它如何影响正常大脑中的其他细胞及其动力学几乎一无所知。这种知识的缺乏在很大程度上是由于体外方法无法准确再现小胶质细胞高度敏感和适应的大脑环境，以及缺乏适当的工具来监测和操纵体内小胶质细胞。为了开始克服这些限制，我们已经开发了一套荧光染色和遗传操作，成像和数据分析工具，用于正常大脑中细胞动力学的研究。例如，这些工具使我们能够揭示行为小鼠中星形胶质细胞和神经元兴奋的新形式以及健康成人大脑中小胶质细胞的监视行为。在这些方法的基础上，我们现在建议开发一套新的工具，用于对大脑浅层和深层结构中的小胶质细胞进行微创监测和操作。这将使我们能够对这些细胞的功能动力学和相互作用提供前所未有的见解。因此，我们的研究对我们对小胶质细胞的看法，它们在健康和患病大脑中的有益和有害作用，以及更普遍地对我们揭示正常大脑中复杂的细胞-细胞相互作用的能力具有广泛的影响。 公共卫生相关性：小胶质细胞参与基本上所有脑部病变的发作，进展和/或消退。然而，人们对它们在完整环境中的性质知之甚少。 个脑袋我们建议通过开发和应用新的研究工具来填补我们知识中的这一重要空白。

DESCRIPTION (Provided by the applicant) Abstract: Microglia are key players in most if not all central nervous system (CNS) disorders including neuropathic pain, Rett syndrome, amyotrophic lateral sclerosis and Nasu-Hakola disease. Recent evidence suggests that these cells also participate in regulation of neurogenesis, circuit remodeling and synaptic plasticity during normal postnatal development, influences they may continue to exert in the adult brain. Despite these vital roles in normal physiology and pathology, we know virtually nothing about these cells' signaling, how it is influenced by local environment and disease, and how it influences other cells and their dynamics in the normal brain. This dearth of knowledge is largely due the inability of in vitro approaches to accurately recapitulate the brain's environment to which microglia are highly sensitive and adaptable and a lack of proper tools to monitor and manipulate microglial cells in vivo. To begin to overcome these limitations we have developed a set of fluorescence staining and genetic manipulation, imaging and data analysis tools for the study of cellular dynamics in the normal brain. These tools have allowed us to reveal, for example, novel forms of astrocytic and neuronal excitation in behaving mice and microglia's surveillance behavior in the healthy adult brain. Building on these approaches we now propose to develop a new set of tools for minimally invasive monitoring and manipulation of microglia in both superficial and deep brain structures. This will allow us to deliver unprecedented insight into these cell's functional dynamics and interactions. Our research therefore has broad implications for our view of microglia, their beneficial and detrimental roles in the healthy and diseased brain and more generally for our ability to uncover complex cell-cell interactions in the normal brain. Public Health Relevance: Microglial cells are involved in onset, progression and/or resolution of essentially all brain pathologies. However, little is known about their properties in the intact brain. We propose to fill this essential gap in our knowledge through development and application of novel research tools.