Mechanisms of Translation in the CNS

中枢神经系统的翻译机制

• 批准号: 8606520
• 负责人: Joel D Richter
• 金额: $ 42.83万
• 依托单位: UNIV OF MASSACHUSETTS MED SCH WORCESTER
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-02-01 至 2018-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8606520
• 关键词: 3' Untranslated Regions; Aging; Alzheimer' s Disease; Animals; Autistic Disorder; Axon; Behavior; Binding; Binding Proteins; Brain; Cognitive; Communication; Complex; Data; Defect; Dementia; Dendrites; Disease; Enzymes; Foundations; Fragile X Syndrome; Fright; Genetic Translation; Goals; Health; Hippocampus (Brain); Human; In Vitro; Knockout Mice; Learning; Length; Life; Link; Long-Term Potentiation; Mediating; Memory; Memory Loss; Messenger RNA; Methods; Molecular; Mus; Neuraxis; Neurites; Neurons; Neuropathy; Neurophysiology - biologic function; Neuropil; Neurotransmitter Receptor; Parkinson Disease; Poly A; Poly(A) Tail; Polyadenylation; Polyadenylation Pathway; Polynucleotide Adenylyltransferase; Process; Proteins; RNA-Binding Proteins; Rodent; Scaffolding Protein; Schizophrenia; Set protein; Site; Slice; Synapses; Synaptic plasticity; Testing; Translating; Translations; deep sequencing; in vivo; inhibitor/antagonist; nervous system disorder; postsynaptic; public health relevance; research study; response; scaffold; synaptic function

DESCRIPTION (provided by applicant): The broad objective of this proposal is to delineate mechanisms of mRNA translation in mammalian neurons and especially dendrites that modify synaptic efficacy. CPEB, a sequence-specific mRNA binding protein that promotes cytoplasmic polyadenylation- induced translation, is present at synapto-dendrites of mammalian neurons. CPEB knockout mice display defects in synaptic plasticity and learning and memory, indicating the importance of the cytoplasmic polyadenylation machinery in complex brain function. CPEB nucleates a set of factors on mRNA to promote polyadenylation including the non- canonical poly(A) polymerase Gld2, the deadenylating enzyme PARN, the eIF4E-binding protein neuroguidin (Ngd), the scaffold protein symplekin, and others. These proteins reside in a complex in dendrites of mammalian neurons where they modulate the polyadenylation and translation of several mRNAs. Two of these factors, Gld2 and Ngd, regulate synaptic plasticity in hippocampal neurons but do so in opposite directions; Gld2 depletion induces a deficit in long-term potentiation (LTP) while Ngd depletion enhances it. Moreover, Gld2 depletion reduces translation in dendrites while Ngd depletion stimulates it. These data indicate that the interplay among CPEB, Gld2, and Ngd form a coherent molecular foundation of translation control in dendrites that in turn modulates synaptic efficacy. The goals of the first specific aim are to identify deadenylating enzymes that are likely to modify poly(A) length and changes in translation as well as to assess whether they influence synaptic function. Aim 2 is to investigate the full panoply of mRNAs that are bound by CPEB in the brain and determine whether they undergo activity-dependent polyadenylation and translation in dendrites. The goal of aim 3 is to develop and use a new deep sequencing method to identify dendritic mRNAs that undergo cytoplasmic polyadenylation and translation in response to in LTP induction in vitro and learning in vivo. These experiments will enhance our understanding of how local mRNA translation in neurons mediates synapse function, which has important implications for higher brain function and neuropathies such as autism, Alzheimer's Disease, Parkinson's Disease, and others.

描述（由申请人提供）：该提案的广泛目标是描述哺乳动物神经元中mRNA翻译的机制，尤其是修改突触功效的树突。 CPEB是一种促进细胞质聚腺苷酸化诱导翻译的序列特异性mRNA结合蛋白，存在于哺乳动物神经元的突触神经化石上。 CPEB敲除小鼠在突触可塑性和学习和记忆中表现出缺陷，表明细胞质聚腺苷酸化机制在复杂的大脑功能中的重要性。 CPEB对mRNA上的一组因子构成了促进聚腺苷酸化的一组因子，包括非典型聚（A）聚合酶GLD2，deadenylation酶PARN，EIF4E结合蛋白神经蛋白（NGD），caffold蛋白蛋白质蛋白Symplekin等。这些蛋白质驻留在哺乳动物神经元树突中的一个复合物中，它们调节了几种mRNA的聚腺苷酸化和翻译。其中两个因素GLD2和NGD调节海马神经元中的突触可塑性，但在相反的方向上执行。 GLD2耗竭诱导长期增强（LTP）的赤字，而NGD耗竭会增强它。此外，GLD2耗竭可减少树突中的翻译，而NGD耗竭会刺激它。这些数据表明，CPEB，GLD2和NGD之间的相互作用形成了树突中翻译控制的一致分子基础，而树突中则调节了突触功效。第一个特定目的的目标是鉴定可能修改poly（a）长度和翻译变化以及评估它们是否影响突触功能的去甲基化酶。 AIM 2是研究由CPEB在大脑中绑定的mRNA的完整全盘，并确定它们是否在树突中经历活性依赖性的聚腺苷酸化和翻译。 AIM 3的目的是开发和使用一种新的深层测序方法来鉴定在体外诱导LTP诱导和体内学习的LTP诱导中经历细胞质聚腺苷酸化和翻译的树突mRNA。这些实验将增强我们对神经元中局部mRNA翻译如何介导突触功能的理解，这对较高的大脑功能和神经病具有重要意义，例如自闭症，阿尔茨海默氏病，帕金森氏病等。