Optogenetic signaling inhibitors for studying brain plasticity

项目总结/摘要 突触可塑性被认为是大脑学习和记忆的基础。信令 突触和行为可塑性的潜在机制已被广泛研究 借助药理学和遗传学对信号的操纵。但却 一直难以评估信号活动的时空方面。的 这项提案的目标是开发一种基于基因编码光的新技术， 诱导激酶抑制剂，以解决所需的信号传导的时空动力学 突触和行为的可塑性。我们的初步数据使用了一种新的- 开发的光诱导CaMKII抑制剂表明，约60 s的CaMKII活性是 足以诱导树突棘的结构可塑性。我们的目标是1）开发照片- 包括CaMKII、PKC和PKA在内的激酶的诱导型抑制剂，2）确定时机 的激酶活性所需的突触可塑性切片，和3）确定 学习相关树突棘所需激酶活性时空窗口 体内周转和动物学习后的性能改善。新工具将 提供了新的见解激酶在体内的行动，在突触和行为 可塑性。

Project Summary/Abstract Synaptic plasticity is thought to be a basis of learning and memory of the brain. Signaling mechanisms underlying synaptic and behavioral plasticity have been extensively studied with the aid of pharmacological and genetic manipulation of signaling. However, it has been difficult to assess the spatiotemporal aspects of signaling activity particularly. The goal of this proposal is to develop a new technique based on genetically encoded light- inducible kinase inhibitors to resolve the spatiotemporal dynamics of signaling required for synaptic and behavioral plasticity in vivo. Our preliminary data using a newly- developed photo-inducible CaMKII inhibitor demonstrates that ~60 s of CaMKII activity is sufficient to induce structural plasticity of dendritic spines. We aim to 1) develop photo- inducible inhibitors for kinases including CaMKII, PKC and PKA, 2) determine the timing of kinase activity required for synaptic plasticity in slices, and 3) identify the spatiotemporal window of kinase activity required for learning-related dendritic spine turnover in vivo and animal’s performance improvement after learning. The new tool will provide new insights into the action of kinases in vivo during synaptic and behavioral plasticity.