Imaging neuronal activity with voltage-sensitive GFP

使用电压敏感 GFP 成像神经元活动

• 批准号: 7120191
• 负责人: SHAWN R LOCKERY
• 金额: $ 12.11万
• 依托单位: UNIVERSITY OF OREGON
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-09-01 至 2008-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7120191
• 关键词: Caenorhabditis elegans; Xenopus; behavioral /social science research tag; bioimaging /biomedical imaging; biotechnology; chemotaxis; ethology; fluorimetry; green fluorescent proteins; neuroimaging; neurons; protein engineering; protein structure function; space perception; voltage /patch clamp

DESCRIPTION (provided by applicant): The long-term goal of the candidate's research program is to understand as completely as possible the neuronal basis of the entire behavioral repertoire of a simple invertebrate--the nematode worm C. elegans. To this end he has developed several important new techniques that have advanced the research goals of his and other labs. However, achieving a virtually complete account of behavior will require another new technique in C. elegans: optical recording of membrane potential from multiple neurons in intact, behaving animals. The candidate seeks release from teaching and reduced administration in order to develop molecular probes for membrane potential based on green fluorescent protein (GFP), and to use these probes to further the goals of his continuing research on the neuronal basis of spatial orientation behavior in C. elegans. The career development plan has four components. Component I is to coordinate and participate in a new consortium of three PI's and six consultants to re-engineer the FlaSh protein, a promising form of voltage-dependent GFP. The consortium brings together world leaders in structural biology, biophysics, fluorescent probes, and optical recording with the aim of enhancing the speed and sensitivity of the original FlaSh molecule. If successful, the re-engineered protein will be used to elucidate the neuronal basis of chemotaxis, a widespread form of spatial orientation, in the nematode C. elegans. The proposed research is significant because it will likely lead to new genetically targetable probes of neuronal activity that can be used in many other important experimental systems. In addition, this work will deepen our understanding of neural computations underlying a common goal-directed behavior. Components II and III are, respectively, technical courses and tutorial visits to laboratories at the leading edge of optical recording technology. These two activities will give the PI new technical background needed to complete the work of Component I. Component IV is a course the PI will teach in research ethics, drawing on his extensive background in analytical philosophy.

描述（由申请人提供）：候选人研究计划的长期目标是尽可能完全理解简单无脊椎动物的整个行为库的神经元基础 - 线虫蠕虫秀丽隐杆线虫。为此，他开发了几种重要的新技术，这些技术推动了他和其他实验室的研究目标。但是，实现几乎完整的行为说明将需要秀丽隐杆线虫中的另一种新技术：从完整，行为动物中多个神经元中膜电位的光学记录。候选人寻求从教学和减少给药中释放，以开发基于绿色荧光蛋白（GFP）的膜电位的分子探针，并利用这些探针进一步进一步研究了他对C. exlelans中空间取向行为的神经元基础的持续研究的目标。 职业发展计划有四个组成部分。组件I将协调并参与一个由三个PI和六名顾问组成的新财团，以重新设计闪光蛋白，这是电压依赖性GFP的有前途的形式。该财团将结构生物学，生物物理学，荧光探针和光学记录的世界领导者汇集在一起​​，以增强原始闪光分子的速度和灵敏度。如果成功，则重新设计的蛋白将用于阐明e秀素e秀隐杆线虫中趋化性的神经元基础，这是一种广泛的空间取向形式。拟议的研究很重要，因为它可能会导致新的遗传靶向探针神经元活性，这些探针可用于许多其他重要的实验系统。此外，这项工作将加深我们对共同目标指导行为基础的神经计算的理解。组件II和III分别是对光学记录技术领先优势实验室的技术课程和教程访问。这两项活动将为完成组件I的工作所需的PI新技术背景。组件IV是PI在研究伦理学中教授的课程，借鉴了他在分析哲学方面的广泛背景。