A Genetic Strategy for Altering Neural Circuitry in Drosophila

描述（由申请人提供）：神经回路提供支持行为的物理基础设施，神经科学的主要目标是了解回路如何控制行为。目前，破坏完整动物体内电路功能的遗传策略集中在改变单细胞的功能上。然而，大多数神经元与多个细胞突触，目前还没有专门改变神经元接触的一个子集的通用策略。该项目的目标是创建一个遗传系统，用于故意破坏神经元连接的一个子集，允许在单个接触或单个突触的水平上解剖神经回路。我们建议在果蝇中实现我们的目标，通过创造人工工程的生长锥运动调节器，可以用来有意雕刻神经元连接的模式。该项目的最初目标是表征一系列人工工程配体/受体对，以确定哪些能够影响果蝇的生长锥导航，而不会产生意外的副作用。该项目的第二个目的是表征这些选定的配体/受体对改变单独识别的神经元与其识别的靶点的连接性的能力，使用组织学和电生理学检查结果。该项目的最终目的是将配体/受体系统应用于一个良好表征的学习和记忆神经回路，使用配体/受体系统来剖析神经回路内特定连接的功能重要性。这项工作中开发的工具将立即被其他研究人员用于操纵果蝇的各种神经回路。这里创建的工具可以用于其他细胞类型的其他目的，例如破坏细胞迁移或细胞接触。此外，虽然这一建议将提供苍蝇的原则证明的发展，在这里开发的策略的变化应该是可行的哺乳动物，并可以作为实验和工程工具的应用。神经系统的布线模式为人类行为提供了物理框架，包括与人类健康相关的基本过程，如学习和记忆。该提案提出开发新的方法来操纵神经系统的布线，以研究它如何控制行为。这些发展将导致对学习和记忆的新见解，并且该资助中开发的基本策略的潜在应用可用于改变用于治疗目的的连接。

DESCRIPTION (provided by applicant): Neural circuits provide the physical infrastructure that supports behavior, and a major goal in neuroscience is to understand how circuits function to control behavior. At present, genetic strategies for disrupting circuit function within an intact animal focus on altering the function of single cells. However, most neurons synapse with multiple cells, and there is currently no general strategy for specifically altering just a subset of a neuron's contacts. The goal of this project is to create a genetic system for intentionally disrupting a subset of a neuron's connections, permitting the dissection of neural circuits at the level of the single contact or the single synapse. We propose to achieve our goal in Drosophila by creating artificially engineered regulators of growth cone movement that can be used to intentionally sculpt patterns of neuronal connectivity. The initial aim of the project involves characterizing a series of artificially engineered ligand/receptor pairs to determine which are able to influence growth cone navigation in the fly without unintended side effects. The second aim of the project involves characterizing the ability of these selected ligand/receptor pairs to alter the connectivity of an individually identified neuron to its identified targets, examining the consequences using histology and electrophysiology. The final aim of the project involves applying the ligand/receptor system to a well- characterized neural circuit for learning and memory, using the ligand/receptor system to dissect the functional importance of specific connections within the neural circuit. The tools developed in this work will be immediately usable by other investigators to manipulate various neural circuits in Drosophila. The tools created here could be used in other cell types for other purposes, such as the disruption of cell migration or cell contact. In addition, while this proposal will provide proof-of-principle development in the fly, variations on the strategy developed here should be feasible in mammals and could have applications as both experimental and engineering tools. The wiring pattern of the nervous system provides the physical framework for human behavior, including such fundamental processes of relevance to human health such as learning and memory. This proposal proposes to develop new ways to manipulate the wiring of the nervous system in order to study how it controls behavior. These developments will lead to new insights into learning and memory and potential applications of the basic strategy developed in this grant could be used to alter connections for therapeutic purposes.