Imaging neuronal activity with voltage-sensitive GFP

描述（由申请人提供）：候选人研究项目的长期目标是尽可能完整地理解一种简单无脊椎动物——线虫C. elegans的整个行为机制的神经元基础。为此，他开发了几项重要的新技术，推动了他和其他实验室的研究目标。然而，要对秀丽隐杆线虫的行为进行几乎完整的描述，还需要另一项新技术：对完整的、行为正常的动物的多个神经元的膜电位进行光学记录。该候选人寻求从教学和减少管理中解放出来，以开发基于绿色荧光蛋白（GFP）的膜电位分子探针，并利用这些探针进一步实现他对秀丽隐杆线虫空间取向行为的神经元基础的持续研究目标。

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.