TRANSCRIPTION FACTOR/REGULATION IN LEARNING AND MEMORY

学习和记忆中的转录因子/调节

• 批准号: 6472798
• 负责人: SANDRA PENA DE ORTIZ
• 金额: $ 11.03万
• 依托单位: UNIVERSITY OF PUERTO RICO RIO PIEDRAS
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-07-01 至 2002-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6472798
• 关键词: antisense nucleic acid; behavioral /social science research tag; body movement; gene expression; gene induction /repression; genetic regulation; immunocytochemistry; laboratory rat; learning; long term memory; memory; microarray technology; nuclear receptors; nucleic acid probes; oligonucleotides; orientation; regulatory gene; space perception; transcription factor; western blottings

Description (Adapted from Application): The overall goal of this research is to examine the role of transcriptional gene regulation in acquisition and retention of spatial memory. Spatial memory is associative in nature and relates to the orientation and movement of an individual in space in relation to the location of objects in the surrounding environment. A basic understanding of how spatial memories are formed will enable the design of appropriate and effective treatment and prevention strategies for the various neurologic and neuropsychiatric conditions that display alterations in memory processing, such as Alzheimer's disease (AD) and stroke. Regulation of gene expression is considered essential for the establishment of long-lasting cellular changes that underlie the formation of long-term memory. The hzf-3/nurr1 gene is a member of the family of inducible orphan nuclear receptors (ONRs), comprised by immediate-early transcription factors thought to be involved in synaptic plasticity. Recent studies by the PI have shown that the mRNA levels of hzf-3 increase in the rat hippocampus following induction of long-term potentiation at the hippocampal mossy fiber-CA3 synapse and during acquisition of the hole board food search task for spatial memory. Results suggest that the expression of the hzf-3 gene in the hippocampus is closely associated with acquisition and retention of spatial learning. The investigators hypothesize that the hzf-3 protein product is required for normal learning acquisition and that it regulates the transcription of genes whose control is important for appropriate spatial memory formation. This proposal will examine the role of the hzf-3/nurr1 protein product in the formation of long-term spatial memory and will identify genes expressed during acquisition and retention of spatial memory including those that require the hzf-3 product for transcriptional activation. Specific aim 1 will examine the effects of sequence specific antisense oligodeoxynucleotides, designed to block translation of the hzf-3 protein product, on the acquisition of the hole board food search task for spatial memory. Oligodeoxynucleotides are microinfused intrahippocampally prior to training in the hole board food search task and the effects on acquisition and retention are determined. The effectiveness of translational disruption of the hzf-3/nurr1 gene will be determined with Western blots and immunohistochemistry and compared with the behavioral effects of the hzf-3 antisense oligodeoxynucleotides. In specific aim 2, cDNA microarrays are used to define a transcripts expression profile of spatial learning in the Hole Board Food Search Task. Commercially available cDNA microarrays used initially to select candidate learning and memory transcripts will be used to prepare macroarrays on glass slides that will be used for further analysis and validation. The arrays are hybridized with rat hippocampal cDNA probes prepared from rats trained in the food search maze. Finally, studies in specific aim 3 will define an hzf-3 dependent transcriptional profile of spatial learning by combining antisense treatments with cDNA microarray analysis. DNA arrays will be hybridized with cDNA probes prepared from animals trained in the maze after hippocampal injection of either saline, hzf-3 antisense or control oligodeoxynucleotides. The proposed set of experiments will provide new and fundamental knowledge which concerns the molecular mechanisms subserving information storage in the brain and which is necessary for the development of proper management strategies of neural conditions that display problems in memory processing.

描述（根据应用程序改编）：这项研究的总体目标是 检查转录基因调节在获取和 保留空间记忆。空间记忆本质上是关联的， 与个人在空间中的方向和运动有关 到周围环境中物体的位置。基本 了解如何形成空间记忆将使您能够设计 适当有效的治疗和预防策略 在记忆中显示改变的神经系统和神经精神疾病 加工，例如阿尔茨海默氏病（AD）和中风。基因的调节 表达对于建立持久是必不可少的 细胞变化是长期记忆形成的基础。这 HZF-3/Nurr1基因是诱导孤儿家族的成员 受体（ONR），由即时的转录因子组成 参与突触可塑性。 PI的最新研究表明 诱导后，HZF-3的mRNA水平增加了大鼠海马 海马苔藓纤维-CA3突触和期间的长期增强 获取空间记忆的孔板食品搜索任务。结果 表明HZF-3基因在海马中的表达紧密 与空间学习的获取和保留有关。这 研究人员假设正常需要HZF-3蛋白产物 学习获取，并调节其基因的转录 控制对于适当的空间内存形成很重要。这个建议 将检查HZF-3/Nurr1蛋白产物在形成中的作用 长期空间记忆，并将识别在获取过程中表达的基因 并保留空间内存，包括需要HZF-3产品的记忆 用于转录激活。具体目标1将检查 序列特定的反义寡脱氧核苷酸，旨在阻止 HZF-3蛋白产品的翻译，孔板的获取 空间记忆的食物搜索任务。寡脱氧核苷酸被微灌注 在孔板培训之前，在霍姆室内食品搜索任务和 确定对获取和保留的影响。有效性 HZF-3/Nurr1基因的翻译破坏将通过 蛋白质印迹和免疫组织化学，并将其与行为效应进行比较 HZF-3反义寡脱氧核苷酸的。在特定的目标2中，cDNA 微阵列用于定义空间的转录本表达式曲线 在孔板上学习食物搜索任务。市售cDNA 微阵列最初用于选择候选学习和记忆成绩单 将用于在载玻片上准备宏观阵列 进一步的分析和验证。阵列与大鼠海马杂交 cDNA探针由接受食物搜索迷宫训练的大鼠制备。最后， 特定目标3中的研究将定义HZF-3依赖性转录 通过将反义处理与cDNA相结合的空间学习概况 微阵列分析。 DNA阵列将与制备的cDNA探针杂交 从海马注射任何一种盐水后在迷宫中训练的动物， HZF-3反义或对照寡脱氧核苷酸。提议的一组 实验将提供新的和基本的知识，与 分子机制在大脑中占据信息存储，哪个是 制定神经的适当管理策略所必需的 在内存处理中显示问题的条件。