Translational control of LTP

LTP 的翻译控制

• 批准号: 6781025
• 负责人: EMMANUEL Manuel LANDAU
• 金额: $ 38.14万
• 依托单位: ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-08-01 至 2008-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6781025
• 关键词: NMDA receptors; biological signal transduction; confocal scanning microscopy; dendrites; electrophysiology; genetic translation; immunocytochemistry; laboratory rat; long term potentiation; memory; neural plasticity; phosphoproteins; phosphorylation; phosphotransferases; protein biosynthesis; protein kinase; protein localization; ribosomal proteins; sirolimus; synapses; translation factor; western blottings

DESCRIPTION (provided by applicant): Neuronal plasticity is thought to underlie a variety of behavioral processes, including memory formation and addiction. Long-term potentiation (LTP) is a persistent, use-dependent form of neuronal plasticity exhibited by synapses in the hippocampus, a brain structure that has been implicated in memory formation and retrieval. Hippocampal LTP has emerged as the major model system for studying the cellular basis of behavioral memory. Long-lasting forms of LTP require protein synthesis (translation). However, little is known about the relationship between LTP and the regulation of translation. The experiments of this proposal address this issue by determining the role in LTP of several important translation control points, which are regulated through protein phosphorylation. We will use synaptic stimulation to induce persistent LTP in rat hippocampal slices. These slices will then be analyzed with biochemical and immunohistochemical methods to determine the distribution and phosphorylation of proteins that participate in translation initiation. Experiments will test for the activation of ribosomal S6 kinase and the phosphorylation of S6, a protein of the 40S ribosomal subunit that selectively promotes the translation of mRNAs containing a terminal oligopyrimidine (TOP) tract in their 5' untranslated regions. Other studies will investigate the possible involvement of the translation initiation factor elF4E and its regulatory binding partner, 4E-BP, which preferentially regulate the translation of transcripts containing structured 5' ends. The cellular mechanisms underlying the regulation of these initiation factors will be studied by inhibiting enzymes of the translation initiation pathway as well as more upstream signaling pathways. Finally, a set of immunohistochemical experiments will study the distribution of translation-related phosphoproteins in the dendritic region using quantitative confocal microscopy. The results of this proposal will shed light on the mechanisms that participate in drug addiction, as well as learning and memory, by providing new information on the role of translation control in neuronal plasticity.

描述（由申请人提供）：神经元可塑性被认为是各种行为过程的基础，包括记忆形成和成瘾。长期增强（LTP）是一种持续的，使用的神经元可塑性形式，它是海马中突触表现出的，这是一种与记忆形成和检索有关的大脑结构。海马LTP已成为研究行为记忆的细胞基础的主要模型系统。 LTP的长期形式需要蛋白质合成（翻译）。但是，对LTP与翻译调节之间的关系知之甚少。该提案的实验通过确定几个重要翻译控制点的LTP中的作用来解决此问题，这些作用是通过蛋白质磷酸化来调节的。我们将使用突触刺激来诱导大鼠海马切片中的持续LTP。然后，将使用生化和免疫组织化学方法分析这些切片，以确定参与翻译起始的蛋白质的分布和磷酸化。实验将测试核糖体S6激酶的激活和S6的磷酸化，S6是40S核糖体亚基的蛋白质，可有选择地促进含有末期寡咪定（TOP）的mRNA在其5'未转移区域中的转化。其他研究将研究翻译起始因子ELF4E及其调节结合伴侣4E-BP的可能参与，该伴侣优先调节包含结构化5'末端的转录本的翻译。通过抑制翻译起始途径的酶以及更多上游信号通路，将研究这些起始因子调节的细胞机制。最后，一组免疫组织化学实验将使用定量共聚焦显微镜研究树突状区域中与翻译相关的磷蛋白的分布。该提案的结果将通过提供有关转化控制在神经元可塑性中的作用的新信息，从而阐明参与药物成瘾以及学习和记忆的机制。