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，以及回路中依赖于经验的变化如何带来可塑性或行为变化。为了解决这一基本挑战，我们必须首先在功能回路中识别突触连接的神经元，其次，我们必须可视化在回路和行为被诱导改变的条件下，活体动物中连接的神经元。这将使科学家们能够测试神经连接中依赖经验的变化是否构成行为变化的基础。我们的建议的目标是产生工具来可视化和操作突触连接的神经元在活的动物的功能电路。我们建议使用一种独特的策略来实现这一目标，在这种策略中，我们鉴定出交叉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，在那里它将诱导扩增uas驱动的报告因子或感兴趣的基因的表达。

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.