Activity-Dependent Regulation of CaMKII and Synaptic Plasticity

摘要 对生活中重大事件的记忆是使我们成为个体的一部分。CaMKII已被证明是 在脊椎动物和无脊椎动物中，短期记忆(STM)和长期记忆(LTM)都需要。CaMKII 蛋白质在突触处丰富，并在局部合成，以响应导致LTM的活动模式 队形。这两个特征都需要在基因的远端未翻译部分存在特定的序列 CaMKII基因的表达。尽管在CAMKII的本地翻译方面做了大量的工作，但有几个基本问题 仍未得到回复： 当地翻译对躯体因素有要求吗？我们将扰乱两者之间的联系 细胞体和突触，以测试是否需要将体细胞物质运输到突触 活性刺激的CaMKII的局部合成。 指定局部翻译的信息是如何在信使核糖核酸中编码的？使用转基因编码 荧光记者和实时分析新的蛋白质合成，我们将确定哪些序列 CaMKII突触定位和活性依赖翻译所必需的。 读取这些信息的蜂窝组件是什么？我们将做一个生物信息驱动的 在RNA亲和纯化的同时筛选候选基因，以确定调节基础和 可塑性刺激的CaMKII积累。 CaMKII的本地翻译中断如何影响LTM？我们将使用条件基因组编辑 移除指定局部翻译的信使核糖核酸序列或用特定突变体取代它们。我们会 确定成人学习回路中的哪些细胞在LTM形成过程中使用这些信息。 该项目利用尖端的遗传学、细胞生物学和光学方法来解决分子基础 一种系统发育保守的可塑性机制，这将进一步加深我们对 复杂的行为。

Abstract Memories of salient events in our lives are part of what makes us individuals. CaMKII has been shown to be required in both vertebrates and invertebrates for short-term (STM) and long-term memory (LTM). CaMKII protein is enriched at synapses, and is synthesized locally in response to activity patterns that lead to LTM formation. Both of these features require specific sequences present in the distal untranslated part of the CaMKII mRNA. In spite of extensive work on local translation of CaMKII, several fundamental questions remain unanswered: Is there a requirement for somatic factors in local translation? We will disrupt the connection between the cell body and the synapse to test whether transport of somatic material to the synapse is required for activity-stimulated local synthesis of CaMKII. How is the information specifying local translation encoded in mRNA? Using transgenes encoding fluorescent reporters and real-time assays of new protein synthesis, we will determine what sequences are required for CaMKII synaptic localization and activity-dependent translation. What are the cellular components that read this information? We will do a bioinformatically-driven candidate gene screen in parallel with RNA affinity purification to identify proteins that regulate basal and plasticity-stimulated CaMKII accumulation. How does disruption of local translation of CaMKII affect LTM? We will use conditional genome editing to remove mRNA sequences that specify local translation or to replace them with specific mutants. We will determine which cells in the adult learning circuit use this information during LTM formation. This project utilizes cutting-edge genetic, cell biological and optical methods to address the molecular basis of a phylogenetically-conserved mechanism of plasticity in a way that will further our understanding of complex behaviors.