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: 9803208
• 负责人: Leslie C Griffith
• 金额: $ 40.63万
• 依托单位: BRANDEIS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-08-15 至 2023-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9803208
• 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; Genetic Translation; Genomics; Imagery; Individual; Invertebrates; Lead; 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; Translations; Ursidae Family; Vertebrates; Work; calmodulin-dependent protein kinase II; cell type; experimental study; fast axonal transport; fly; genome editing; immunoreactivity; in vivo; in vivo evaluation; insight; long term memory; mature animal; mutant; postsynaptic; 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.

抽象的 我们生活中显着事件的记忆是使我们个人的一部分。 Camkii已被证明是 在短期（STM）和长期记忆（LTM）中，脊椎动物和无脊椎动物都需要。 camkii 蛋白质在突触时富集，并响应导致LTM的活性模式局部合成 形成。这两个特征都需要在未翻译的部分中存在的特定序列 Camkii mRNA。尽管在Camkii的本地翻译方面进行了广泛的工作，但有几个基本问​​题 保持没有答案： 在局部翻译中是否需要躯体因素？我们将破坏 细胞体和突触测试是否需要将躯体材料传输到突触 活动刺激的CAMKII局部合成。 指定局部翻译的信息如何在mRNA中编码？使用转基因编码 荧光记者和新蛋白质合成的实时测定，我们将确定哪些序列是 CAMKII突触定位和活动依赖性翻译所必需的。 读取此信息的细胞组件是什么？我们将进行生物信息驱动 候选基因筛选与RNA亲和力纯化并行，以鉴定调节基础和基础的蛋白质 可塑性刺激的CAMKII积累。 CAMKII局部翻译的破坏如何影响LTM？我们将使用条件基因组编辑 去除指定局部翻译或用特定突变体代替它们的mRNA序列。我们将 确定成人学习电路中的哪些细胞在LTM形成期间使用此信息。 该项目利用尖端的遗传，细胞生物学和光学方法来解决分子基础 可塑性的系统发育机制的方式，这将进一步了解我们对 复杂的行为。