BRAIN EAGER: Development of Robotic Microscopy to monitor the longitudinal molecular dynamics of single neurons and circuits in situ in mammalian brain

BRAIN EAGER：开发机器人显微镜来监测哺乳动物大脑中单个神经元和回路的纵向分子动力学

• 批准号: 1451350
• 负责人: Steven Finkbeiner
• 金额: $ 30万
• 依托单位: The J. David Gladstone Institutes
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-09-01 至 2016-08-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1451350&HistoricalAwards=false
• 关键词: BRAIN; EAGER; Development; Robotic; Microscopy

This project is directed at developing a novel technology, robotic microscopy (RM), to "bridge multiple spatial, temporal, and organizational scales to provide fundamental insights into the emergent properties of neural circuitry that ultimately lead to behavior and cognition." There are two primary benefits of the proposed studies. First, the experiments will result in the development of an RM instrument that can monitor the molecular dynamics of individual neurons in slices of living brain tissue over weeks at a time. Second, the experiments will use RM to study for the first time how learning and memory change the fundamental properties of specific neurons in situ to enhance synaptic plasticity, providing new insights into the mechanisms involved in learning and memory. In the past, RM was used to study neurons in culture. The primary activity in the proposed studies will be to validate the utility of RM to monitor neuronal dynamics in brain slices that maintain their natural physiological connectivity and architecture. Other benefits will be access of the scientific community to a technology that can examine the biochemistry of neurons and other cell types longitudinally and the development of new instrumentation that can be used in academic courses that teach students, postdoctoral fellows and research scientists novel imaging approaches to study brain circuits.The problem to be addressed is whether RM can be used to study neurons expressing the Arc gene in situ in hippocampal brain slices. Arc is important for long-term memory consolidation and synaptic plasticity, and its activity is greatly enhanced in specific neuron populations by stimuli that affect learning. The methods to be employed will involve the use of novel Arc genetic probes and transgenic mice to determine if RM can identify selective Arc-expressing neurons activated in brain slices by long-term potentiation (LTP) and long-term depression (LTD) or in brain in vivo during learning and memory consolidation. The goals of the studies will be to determine if different Arc neuronal circuits mediate LTP and LTD, and if the same Arc expressing neurons activated in vivo in the brain during learning can be monitored by RM in situ in brain slices. The scope of the studies will determine if RM can identify and study different Arc neuronal circuits activated in vitro and in vivo by different forms of learning and memory consolidation.

该项目旨在开发一种新技术——机器人显微镜（RM），以“连接多个空间、时间和组织尺度，为最终导致行为和认知的神经回路的新兴特性提供基本见解。”拟议的研究有两个主要好处。首先，这些实验将开发出一种 RM 仪器，可以在数周内一次监测活体脑组织切片中单个神经元的分子动力学。其次，实验将首次利用 RM 研究学习和记忆如何原位改变特定神经元的基本特性以增强突触可塑性，为学习和记忆所涉及的机制提供新的见解。过去，RM 用于研究培养物中的神经元。拟议研究的主要活动将是验证 RM 在监测大脑切片中维持其自然生理连接和结构的神经元动态方面的效用。其他好处将是科学界获得一种可以纵向检查神经元和其他细胞类型的生物化学的技术，以及开发可用于学术课程的新仪器，这些课程向学生、博士后研究员和研究科学家教授研究大脑回路的新颖成像方法。需要解决的问题是 RM 是否可以用于研究海马脑切片中原位表达 Arc 基因的神经元。弧对于长期记忆巩固和突触可塑性很重要，并且它的活性在特定神经元群体中通过影响学习的刺激而大大增强。所采用的方法将涉及使用新型 Arc 基因探针和转基因小鼠来确定 RM 是否能够识别大脑切片中通过长时程增强 (LTP) 和长期抑制 (LTD) 激活的选择性 Arc 表达神经元，或在学习和记忆巩固过程中体内大脑中激活的选择性 Arc 表达神经元。研究的目标是确定不同的 Arc 神经元回路是否介导 LTP 和 LTD，以及学习期间大脑体内激活的相同 Arc 表达神经元是否可以通过脑切片中的原位 RM 进行监测。研究范围将决定 RM 是否能够识别和研究通过不同形式的学习和记忆巩固在体外和体内激活的不同 Arc 神经元回路。