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。 我们的方法通过开发新技术来解决蛋白质合成依赖性突触可塑性：单突触和突触翻译分析，从而挑战了当前对突触RNA的定量识别的关注。 这些将使我们能够从两个新的叠加角度重新定义问题：需要在特定的突触中识别受调节的RNA-蛋白质复合物，并需要定义它们在翻译调节中的作用。 将研究具体的突触： 小脑Purkinje神经元（运动部位），CA1锥体神经元的顶部树突（在海马分子层（缔合记忆的位点）和V层V层锥体神经元的视觉皮质（活性相关塑料的位点））中的CA1锥体神经元（缔合记忆的位点）。 这些突触中要研究的关键RNA蛋白复合物将是Argona-MRNA-MIRNA三元络合物，翻译调节的FMRP-MRNA复合物，以及神经元特异性RNA调节蛋白-MRNA复合物已知已知的蛋白质-MRNA络合物存在于Dendrite中，并且存在于Dendrite中，并与3'Utrs（nelavls neelavls neelavl（nelavls）（Hu proteans）（Hu protean）（neelavls）（n Noga） 这些复合物将与同一树突中存在的所有核糖体-MRNA突触复合物的描述进行比较，从而使我们能够通过识别翻译调节的突触mRNA（突触翻译分析）来验证相互作用。 受调节的树突状RNA将通过在两个蛋白质合成的两个良好研究范式中评估其转化状态 - 依赖性突触可塑性：在海马中以及在黑暗饲养后的视觉皮层和后续光线暴露后。 这些研究将彻底改变我们对记忆基础的局部突触mRNA的性质和调节的理解，为对记忆的神经系统疾病（例如阿尔茨海默氏病和其他神经退行性疾病）的神经系统疾病奠定了基础。 公共卫生相关性：RNA的失调 - 基因组DNA和蛋白质之间的关键分子 - 越来越认识到人类神经疾病的根源。 更确切地说，总体上，记忆和复杂的认知功能被认为取决于神经元连接部位蛋白质合成的调节。 了解该法规的机制将在此处进行一系列创新的实验，并且是理解记忆和认知障碍的关键。