Imaging neuronal activity with voltage-sensitive GFP

使用电压敏感 GFP 成像神经元活动

• 批准号: 6611666
• 负责人: SHAWN R LOCKERY
• 金额: $ 11.08万
• 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/6611666
• 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整地了解简单无脊椎动物-线虫C的整个行为库的神经元基础。优美的为此，他开发了几种重要的新技术，推进了他和其他实验室的研究目标。然而，实现行为的实际完整描述将需要C中的另一种新技术。elegans：光学记录膜电位从多个神经元在完整的，行为动物。候选人寻求从教学和减少管理中解脱出来，以开发基于绿色荧光蛋白（GFP）的膜电位分子探针，并使用这些探针进一步实现他在C.优美的 职业发展计划有四个组成部分。第一部分是协调和参与一个由三名PI和六名顾问组成的新联盟，以重新设计FlaSh蛋白，这是一种有前途的电压依赖性GFP形式。该联盟汇集了结构生物学、生物物理学、荧光探针和光学记录领域的世界领导者，旨在提高原始FlaSh分子的速度和灵敏度。如果成功的话，重组的蛋白质将被用于阐明线虫C中趋化性的神经基础，趋化性是一种广泛存在的空间定向形式。优雅的这项拟议中的研究意义重大，因为它可能会导致新的神经元活动的遗传靶向探针，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导向行为背后的神经计算的理解。第二部分和第三部分分别是技术课程和对光学记录技术前沿实验室的辅导访问。这两项活动将为PI提供完成第一部分工作所需的新技术背景。第四部分是PI将教授研究伦理学的课程，利用他在分析哲学方面的广泛背景。