TRANSCRIPTION FACTOR/REGULATION IN LEARNING AND MEMORY

描述（改编自应用）：本研究的总体目标是 研究转录基因调控在收购中的作用， 保持空间记忆。空间记忆本质上是联想的， 与个人在空间中的方位和运动有关， 物体在周围环境中的位置。一个基本 了解空间记忆是如何形成的将使设计 适当和有效的治疗和预防战略， 显示记忆改变的神经学和神经精神学病症 加工，如阿尔茨海默病（AD）和中风。调节基因 表达被认为是建立持久的 长期记忆形成的细胞变化。的 hzf-3/nurr 1基因是可诱导孤儿核转录因子家族的一员， 受体（ONRs），由被认为是 参与突触可塑性。PI最近的研究表明， 诱导后大鼠海马hzf-3 mRNA水平增加， 海马苔藓纤维-CA 3突触的长时程增强， 采集孔板食物搜索任务的空间记忆。结果 提示hzf-3基因在海马中的表达与脑缺血再灌注损伤密切相关， 与空间学习的获得和保持有关。的 研究者推测，hzf-3蛋白产物是正常的 它调节基因的转录， 控制对于适当的空间记忆形成是重要的。这项建议 将研究hzf-3/nurr 1蛋白产物在形成 长期的空间记忆，并将识别基因表达过程中收购 以及空间记忆的保持，包括那些需要HZF-3产品的空间记忆 用于转录激活。具体目标1将审查 序列特异性反义寡脱氧核苷酸，设计用于阻断 翻译的hzf-3蛋白产物，关于孔板的获取 食物搜索任务的空间记忆。微量输注寡脱氧核苷酸 在进行洞板食物搜索任务训练之前， 确定对获得和保持的影响。的有效性 将用以下方法测定hzf-3/nurr 1基因的翻译破坏： 免疫印迹和免疫组化检测并与行为学效应比较 hzf-3反义寡核苷酸。在具体目标2中， 微阵列用于定义转录本的空间表达谱， 在洞板食物搜索任务中学习。市售cDNA 最初用于选择候选学习和记忆转录本的微阵列 将用于在载玻片上制备宏阵列， 进一步分析和验证。将该阵列与大鼠海马进行杂交 从在食物搜索迷宫中训练的大鼠制备的cDNA探针。最后， 在特定目标3中的研究将定义一种HZF-3依赖性转录因子， 联合反义处理和cDNA的空间学习能力 微阵列分析。DNA阵列将与制备的cDNA探针杂交 来自海马注射盐水后在迷宫中训练的动物， HGF-3反义寡脱氧核苷酸或对照寡脱氧核苷酸。拟议的一套 实验将提供新的和基本的知识， 分子机制有助于大脑中的信息存储， 必要的发展适当的管理战略的神经 显示内存处理问题的条件。

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.