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，神经科学的一个主要目标是了解回路如何发挥作用来控制行为。目前，破坏完整动物体内回路功能的遗传策略主要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子集的通用策略。该项目的目标是创建一种遗传系统，用于故意破坏神经元连接的子集，允许在单一接触或单一突触的水平上解剖神经回路。我们建议通过创建人工设计的生长锥运动调节器来实现我们在果蝇身上的目标，这种调节器可以用来刻意塑造神经元连接的模式。该项目的最初目标包括表征一系列人工工程的配体/受体对，以确定哪些能够在没有意外副作用的情况下影响飞行中的生长锥导航。该项目的第二个目标涉及表征这些选定的配体/受体对改变单个识别的神经元与其识别的目标的连接性的能力，并使用组织学和电生理学检查结果。该项目的最终目标包括将配体/受体系统应用于具有良好特性的学习和记忆神经回路，使用配体/受体系统剖析神经回路中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来操纵果蝇的各种神经回路。这里创建的工具可以用于其他类型的细胞用于其他目的，如中断细胞迁移或细胞接触。此外，虽然这项提案将提供即时的原则证明开发，但在这里开发的策略的变体应该在哺乳动物中是可行的，并可以作为实验和工程工具应用。神经系统的连接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。这项建议建议开发新的方法来操纵神经系统的连接，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，这项赠款中开发的基本策略的潜在应用可能被用于改变连接以达到治疗目的。

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.