Molecular probes to visualize endogenous synaptic proteins in vivo

描述（由申请人提供）：本拨款中提出的研究目的是生成工具，使内源性突触蛋白能够在体内神经元中可视化。之前，我们已经产生了被称为fingr的重组抗体（由mRNA显示产生的纤维连接蛋白体内体），它可以在活神经元中表达，在那里它们以无创和高保真度标记内源性靶蛋白。此外，一个转录调控系统控制着fingr的表达水平，使它们的表达水平与内源性对应物完全相同，从而确保低背景。在这项资助中，我们将产生转基因小鼠，表达识别突触蛋白的荧光蛋白融合的finger。这些小鼠大脑中单个神经元中的突触蛋白可以在体内实时可视化。反过来，这将允许事件在

DESCRIPTION (provided by applicant): The purpose of the research proposed in this grant is to generate tools that will allow endogenous synaptic proteins to be visualized in neurons in vivo. Previously we have generated recombinant antibodies known as FingRs (Fibronectin intrabodies generated with mRNA display) that can be expressed in living neurons where they label endogenous target proteins noninvasively and with high fidelity. Furthermore, a transcriptional regulation system controls the expression levels of FingRs so that they are expressed at precisely the same level as their endogenous counterparts, insuring low background. In this grant we will generate transgenic mice that express fluorescent protein-fused FingRs that recognize synaptic proteins. Synaptic proteins in individual neurons in the brains of these mice can be visualized in vivo in real time. In turn, this will allow events at the molecular level to be correlated with events at the cellular, circuit and whole animal level. In particular, by mapping the locations and amounts of synaptic proteins in neurons, it will be possible to monitor the strength of both synaptic inputs and outputs in living neurons. The ability to monitor synaptic strength will be very useful for studying diseases associated with aberrant synaptic connectivity including schizophrenia, mental retardation and autism.