Activity-Dependent Regulation of CaMKII and Synaptic Plasticity

CaMKII 和突触可塑性的活动依赖性调节

• 批准号: 10817516
• 负责人: Leslie C Griffith
• 金额: $ 44.46万
• 依托单位: BRANDEIS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-06-01 至 2026-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10817516
• 关键词: 3' Untranslated Regions; Acute; Address; Adult; Affect; Affinity Chromatography; Alleles; Axotomy; Behavior; Behavioral; Binding Proteins; Bioinformatics; Biological; Biological Assay; Brain; Brain Diseases; Candidate Disease Gene; Cells; Chronic; Clustered Regularly Interspaced Short Palindromic Repeats; Communication; Complex; Coupled; Development; Distal; Drosophila genus; Elements; Event; Genes; Genetic; Genetic Recombination; Individual; Invertebrates; Learning; Maps; Memory; Messenger RNA; Metabolism; Molecular; Mus; Mushroom Bodies; Mutate; Neuromuscular Junction; Neurons; Optical Methods; Pattern; Phylogenetic Analysis; Preparation; Process; Protein Biosynthesis; Proteins; Proteome; RNA; RNA Interference; RNA-Binding Proteins; Regulation; Regulatory Pathway; Reporter; Research; Rest; Ribosomes; Role; Side; Specific qualifier value; Structure; Synapses; Synaptic plasticity; System; Testing; Time; Trans-Activators; Transgenes; Translational Regulation; Translations; Vertebrates; Visualization; Work; calmodulin-dependent protein kinase II; cell type; experimental study; fast axonal transport; fly; genome editing; genomic locus; immunoreactivity; in vivo; in vivo evaluation; insight; long term memory; mRNA Translation; mature animal; mutant; postsynaptic; presynaptic; programs; response; synaptic function; tool

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.

摘要 对我们生活中重要事件的记忆是使我们成为个体的一部分。CaMKII已被证明是 脊椎动物和无脊椎动物都需要短期记忆（STM）和长期记忆（LTM）。CaMKII 蛋白质在突触处富集，并且响应于导致LTM的活动模式而在局部合成 阵这两个特征都需要存在于DNA末端非翻译部分的特定序列。 CaMK Ⅱ mRNA。尽管对CaMKII的本地翻译进行了大量工作，但一些基本问题 仍未得到答复： 本土化翻译是否需要身体因素？我们会破坏 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。 mRNA中指定局部翻译的信息是如何编码的？使用转基因编码 荧光报告和新蛋白质合成的实时测定，我们将确定哪些序列是 这是CaMKII突触定位和活性依赖性翻译所必需的。 什么是读取这些信息的细胞成分？我们将做一个生物信息学驱动的 候选基因筛选与RNA亲和纯化平行，以鉴定调节基础和 可塑性刺激的CaMKII积累。 CaMKII局部翻译的中断如何影响LTM？我们将使用条件基因组编辑 去除指定局部翻译的mRNA序列或用特定突变体替换它们。我们将 确定成人学习回路中的哪些细胞在LTM形成期间使用这些信息。 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问题 在某种程度上，这将进一步加深我们对 复杂的行为