Mapping the mechanisms of protein synthesis-dependent synaptic plasticity

绘制蛋白质合成依赖性突触可塑性的机制

• 批准号: 8412332
• 负责人: ROBERT B DARNELL
• 金额: $ 105.66万
• 依托单位: ROCKEFELLER UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-09-30 至 2017-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8412332
• 关键词: 3' Untranslated Regions; Acute; Address; Alzheimer' s Disease; Apical; Behavioral; Behavioral Paradigm; Binding; Binding Proteins; Brain; Cognition; Complex; Coupled; DNA; Dendrites; Disease; Epitopes; FMRP; Generations; Genetic Translation; Genomics; Hippocampus (Brain); Human; Immunofluorescence Microscopy; Light; Maps; Mediating; Memory; Memory Disorders; Messenger RNA; Methods; MicroRNAs; Molecular; Mus; N-Methyl-D-Aspartate Receptors; Nature; Neurodegenerative Disorders; Neurologic; Neurons; Neuropil; Nova antigen; Plant Roots; Poly(A)-Binding Proteins; Process; Protein Biosynthesis; Proteins; RNA; RNA Binding; RNA Cap-Binding Proteins; RNA-Binding Proteins; Recombinants; Regulation; Rest; Ribosomes; Role; Site; Slice; Staging; Synapses; Synaptic plasticity; System; Technology; Transcript; Translational Regulation; Translations; Untranslated Regions; Validation; Visual; Visual Cortex; base; cognitive function; crosslink; dark rearing; density; design; genetic regulatory protein; genome-wide; genome-wide analysis; hippocampal pyramidal neuron; in vivo; innovation; insight; laser capture microdissection; long term memory; morris water maze; motor learning; nano; new technology; protein complex; research study; validation studies

DESCRIPTION (provided by applicant): We propose to change the current approach to understanding the molecular basis of memory. Our approach challenges the current focus on quantitative identification of synaptic RNAs by developing new technologies to address protein synthesis-dependent synaptic plasticity: single- synapse-CLIP and synaptic translational profiling. These will allow us to redefine the problem from two new superimposed perspectives: the need to identify regulated RNA-protein complexes in specific synapses, and the need to define their role in translational regulation. Specific synapses will be studied: the apical dendrites of cerebellar Purkinje neurons (a site of motor learning), of CA1 pyramidal neurons in the stratum moleculare of the hippocampus (a site of associative memory), and of layer V pyramidal neurons of the visual cortex (a site of activity- dependent plasticity). Key RNA-protein complexes to be studied in these synapses will be Argonaute (Ago)-mRNA-miRNA ternary complexes, translationally regulated FMRP-mRNA complexes, and neuron-specific RNA regulatory protein-mRNA complexes known to be present in the dendrite and to bind 3' UTRs (nElavl (Hu proteins), Nova). These complexes will be compared with a delineation of all ribosome-mRNA synaptic complexes present in the same dendrites, allowing us to validate interactions by identifying translationally regulated synaptic mRNAs (synaptic translational profiling). Regulated dendritic RNAs will be further validated by assessing for their translational state in two well-studied paradigms of protein synthesis- dependent synaptic plasticity: that in the hippocampus, and in the visual cortex after dark rearng and subsequent light exposure. These studies will revolutionize our understanding of the nature and regulation of local synaptic mRNAs that underlie memory, setting the stage for new insight into neurologic diseases of memory such as Alzheimer's and other neurodegenerative diseases. PUBLIC HEALTH RELEVANCE: Dysregulation of RNA-the key molecule between genomic DNA and proteins-is increasingly recognized to lie at the root of human neurologic disease. More precisely, memory, and complex cognitive function in general, is thought to depend on the regulation of protein synthesis at the sites of neuronal connections. Understanding the mechanisms governing this regulation will be studied here in an entirely innovative set of experiments, and they hold the key to understanding disorders of memory and cognition.

描述（由申请人提供）：我们建议改变当前理解记忆分子基础的方法。我们的方法挑战了目前对突触rna定量鉴定的关注，通过开发新技术来解决蛋白质合成依赖的突触可塑性：单突触- clip和突触翻译分析。这将允许我们从两个新的重叠的角度重新定义这个问题：需要识别特定突触中受调节的rna -蛋白复合物，需要定义它们在翻译调节中的作用。具体的突触将被研究：