Deciphering Biochemical Networks in Single Dendritic Spines

 描述(申请人提供)：突触可塑性，学习和记忆的细胞基础，是由数百个信号蛋白组成的复杂的生化信号网络调节的。特别是，树突棘、微小突触后的钙依赖信号 发自树突表面的隔室在诱导长时程突触可塑性中起着关键作用。为了了解该网络的工作原理和突触可塑性的机制，在突触可塑性过程中，需要测量单个树突棘中数百个蛋白质在多种操作下的活性。利用先进的基于荧光共振能量转移(FRET)的技术测量了脊椎中蛋白质的活性。然而，到目前为止，只测量了整个网络的一小部分。由于缺乏对信令组件的测量，我们对信令网络的理解受到限制。因此，本项目的目标是建立一个用于信号传感器开发和优化的高通量系统，以及一个用于单个树突棘可塑性过程中成像信号转导的全自动化系统。使用这种高通量成像系统，我们的目标是将数据输出的整体效率提高数量级，产生大量数据集，这些数据集可以进一步分析信令网络中的信息流和网络元素之间的连接。因此，这个项目有望在我们对神经元细胞内信号的理解方面取得重大进展，并为突触可塑性以及最终学习和记忆的机制提供关键的见解。

 DESCRIPTION (provided by applicant): Synaptic plasticity, a cellular basis of learning and memory, is mediated by a complex biochemical signaling network consists of hundreds of signaling proteins. In particular, Ca2+- dependent signaling in dendritic spines, tiny postsynaptic compartments emanating from dendritic surface, plays a key role in the induction of long-term synaptic plasticity. In order to understand the operational principles of this network and the mechanisms underlying synaptic plasticity, the activity of hundreds of proteins under many manipulations needs to be measured in single dendritic spines during synaptic plasticity. The activity of proteins in spines has been measured using advanced fluorescence resonance energy transfer (FRET)-based techniques. However, thus far only a small fraction of the entire network has been measured. Our understanding of signaling networks is limited by this scarcity of measurements for signaling components. Thus, the goal of this project is to establish a high-throughput system for the development and optimization of signaling sensors, and a fully automated system for imaging signal transduction during plasticity in single dendritic spines. Using this high-throughput imaging system, we aim to improve the overall efficiency of data output by orders of magnitude, producing large data sets that could be further analyzed for information flow in the signaling network and connectivity between network elements. Thus, this project is expected to lead to a dramatic advance in our understanding of intracellular signaling in neurons and provide key insights into the mechanisms underlying synaptic plasticity and ultimately learning and memory.