CREB regulation of gene expression in spatial and contextual memory

这项建议的总体目标是表征在啮齿动物大脑中发生的转录水平的变化，在收购和巩固的空间和上下文记忆。空间记忆在本质上是关联的，并且与个体在空间中与周围环境中的物体的位置有关的方向和运动有关。情境记忆建立了周围环境线索和特定事件之间的认知联系。对空间和背景记忆如何形成的基本理解将使我们能够设计适当的治疗和预防策略，用于各种神经和神经精神疾病，如阿尔茨海默病，中风和创伤后应激障碍。越来越多的证据表明，基因转录的调节在记忆的获得和巩固中起着重要作用。编码cAMP反应元件结合蛋白（CREB）的基因的靶向突变和几个基因的原位杂交研究提供了强有力的证据，支持基因转录在长期记忆获得和巩固中的作用。我们的总体假设是，CREB功能是必要的，以介导整个收购和巩固记忆的基因转录的变化。我们认为学习和记忆过程伴随着大脑不同区域内特定基因转录物的积累和下调。此外，我们假设，一个子集的基因调控过程中收购和巩固的空间记忆在Morris水迷宫和孔板食物搜索任务，使用CREB基因敲除小鼠和诱导CREB突变体。含有数千个小鼠cDNA的DNA微阵列将用于杂交实验中，使用33P标记的cDNA探针，所述探针从小鼠脑mRNA中制备，所述小鼠脑mRNA提取自处于空间记忆获得和巩固的不同阶段的动物。Specific Aim 2将监测CREB敲除小鼠和诱导型突变小鼠中数千个小鼠基因的表达，以识别与恐惧相关的长期记忆中CREB调控下的转录物。最后，具体目标3将研究CREB突变小鼠在海马切片中诱导长时程电位后发生的基因表达变化。拟议的一系列实验将提供新的和基本的知识，涉及的分子机制subserving信息存储在大脑中。

The overall goal of this proposal is to characterize the changes in transcript levels which occur in the rodent brain during acquisition and consolidation of spatial and contextual memory. Spatial memory is associative in nature and relatives to the orientation and movement of an individual in space in relationship to the location of objects in the surrounding environment. Contextual memory establishes cognitive associations between surrounding environmental cues and specific events. A basic understanding of how spatial and contextual memories are formed will enable the design of appropriate treatment and prevention strategies for the various neurologic and neuropsychiatric conditions that display alterations in memory processing, such as Alzheimer's disease, stroke and post-traumatic-stress disorder. An expanding body of evidence reveals a role for the regulation of gene transcription in acquisition and consolidation of memory. Targeted mutation of the gene encoding the cAMP Responsive Element Binding protein (CREB) and in situ hybridization studies of several genes provide strong evidence in support of a role of gene transcription in long-term memory acquisition and consolidation. Our overall hypothesis is that CREB function is necessary to mediate changes in gene transcription throughout acquisition and consolidation of memory. We propose that learning and memory processes are accompanied by accumulation and down regulation of specific gene transcripts within different regions of the brain. Moreover, we hypothesize that a subset of the genes regulated during acquisition and consolidation of spatial memory in the Morris Water maze and the hole board food search task using both CREB knockout mice and inducible CREB mutants. DNA microarrays containing thousands of mouse cDNAs will be used in hybridization experiments using 33P labeled cDNA probes prepared from mouse brain mRNA extracted from animals at different stages of acquisition and consolidation of spatial memory. Specific Aim 2 will monitor the expression of thousands of mouse genes in CREB knockout and inducible mutant mice to identify transcripts under CREB regulation in long-term memory associated with fear. Finally, Specific Aim 3 will study the changes in gene expression which occur in CREB mutant mice following induction of long-term potential in hippocampal slices. The proposed set of experiments will provide new and fundamental knowledge which concerns the molecular mechanisms subserving information storage in the brain.