Trans-Synaptic Transport of Endogenous Proteins to Label Circuits

描述（由申请人提供）：神经科学的一个基本挑战是确定神经元回路如何控制行为，以及回路中的经验依赖性变化如何带来可塑性或行为变化。为了解决这一基本挑战，我们必须首先识别功能回路中的突触连接神经元，其次，我们必须在诱导回路和行为发生变化的条件下，可视化活体动物中的连接神经元。这将使科学家们能够测试神经元连接的经验依赖性变化是否是行为变化的基础。我们计划的目标是生成工具来可视化和操纵活体动物功能回路中的突触连接神经元。我们建议使用一种独特的策略来实现这一目标，在该策略中，我们确定跨突触的内源性蛋白质，并使用这些蛋白质将gal4转录激活因子转运到突触后神经元中，在那里它将诱导UAS驱动的报告基因或感兴趣的基因的扩增表达。 公共卫生相关性：在完整的大脑中可视化和操纵神经元回路的能力将是理解大脑回路如何影响行为的一个重大进展。我们建议开发一种新的方法来标记跨突触连接的神经元，这是适合在体内的时间推移成像。

DESCRIPTION (provided by applicant): A fundamental challenge in neuroscience is to determine how neuronal circuits control behavior and how plasticity, or changes in behavior, is brought about by experience- dependent changes in circuitry. To address this basic challenge we must first identify synaptically-connected neurons within a functional circuit and second, we must be to visualize connected neurons in living animals under conditions in which circuits and behavior are induced to change. This will allow scientists to test whether experience- dependent changes in neuronal connections underlie changes in behavior. The goal of our proposal is to generate tools to visualize and manipulate synaptically-connected neurons within functional circuits in living animals. We propose to accomplish this goal using a unique strategy in which we identify endogenous proteins which cross synapses and use these proteins to transport the gal4 transcriptional activator into postsynaptic neurons, where it will induce amplified expression of UAS-driven reporters or genes of interest. PUBLIC HEALTH RELEVANCE: The ability to visualize and manipulate neuronal circuits in the intact brain is a will be a major advance in understanding how brain circuits affect behavior. We propose to develop a novel method to label trans-synaptically connected neurons which is amenable to in vivo time-lapse imaging.