Molecular markers of chronic stress vulnerability/resilience

描述（由申请人提供）：压力在大脑和外周系统中启动一系列生化事件，使生物体能够科普新的和具有挑战性的情况。然而，慢性应激状态的累积效应对大多数生理系统产生负面影响，因为靶细胞长时间暴露于应激介质。人类的研究结果指出，慢性压力与焦虑症、抑郁症和认知障碍之间存在联系，这一发现得到了啮齿动物行为观察的广泛支持。慢性或长期压力的生理和行为后果取决于个体脆弱性的遗传和环境介质。尽管有大量研究调查压力，但脆弱性或弹性的分子机制尚不清楚。这项建议的目的是确定的敏感性和弹性的慢性压力的分子标志物，作为第一步的压力介质的识别。具体目标1）在四种不同的近交系大鼠中，在慢性束缚应激（CRS）范例停止期间和停止之后，确定慢性束缚应激的行为后果。选择这些近交系Wistar京都（WKY）、Fisher 344（F344）、布朗挪威（BN）和刘易斯（LEW）代表对慢性应激的独特易感性和/或恢复力。将采用高架十字迷宫（高架十字迷宫）测试来评估慢性应激对焦虑相关行为反应的影响。具体目的2）使用一种产生假设的、无偏的方法，即AffiliatesDMA微阵列分析，我们将检测来自四个近交系的雄性动物对CRS的反应中的两个脑区（杏仁核和海马）和血液中的基因表达谱变化。将对从暴露于以下条件的动物获得的组织进行微阵列分析：i）无应激，其将用于确定基线基因表达ii.）14天的CRS范例，这将告知我们应激反应的分子标志物，以及iii.）CRS停止后10天，以确定应激后反应的标志物。通过采用微阵列分析的RNA分离的两个大脑区域高度相关的焦虑和压力，从血液中，我们将确定在这些组织中的基因表达的共同概况在慢性应激暴露期间和之后。具体目标3。通过实时荧光定量RT-PCR确定最有希望的候选标记基因。这项探索性研究将确定在特定的大脑区域或血液中对慢性压力做出反应时单独或伴随改变的基因。这些情景将使我们能够从血液中识别出慢性压力的候选标记，血液代表了人类遗传物质的可获得来源。

DESCRIPTION (provided by applicant): Stress initiates a cascade of biochemical events in the brain and peripheral systems that enable the organism to cope with novel and challenging situations. However, the cumulative effects of chronic stress states negatively affects most physiologic systems because the target cells are exposed to the stress mediators over a long period of time. Human findings, pointing to a linkage between chronic stress and anxiety disorders, depression and cognitive impairments, have been extensively supported by behavioral observations in rodents. The physiological and behavioral consequences of chronic or prolonged stress depend on genetic and environmental mediators of the individual's vulnerability. The molecular mechanisms of vulnerability or resilience are not known despite the large number of studies investigating stress. This proposal aims to determine the molecular markers of sensitivity and resilience to chronic stress as a first step towards the identification of stress mediators. The aims of this proposal are: Specific Aim 1) Determine the behavioral consequences of chronic restraint stress, during and after the cessation of a chronic restrain stress (CRS) paradigm in four different inbred strains of rats. These inbred strains, the Wistar Kyoto (WKY), Fisher 344 (F344), Brown Norway (BN) and Lewis (LEW), are chosen to represent distinctive susceptibility and/or resilience to chronic stress. The elevated plus-maze (EPM) test will be employed to assess the effect of chronic stress on anxiety-related behavioral responses. Specific Aim 2) Using a hypothesis-generating, unbiased approach, the Affimetrix DMA microarray analysis, we will examine the gene expression profile changes in two brain areas, the amygdale and the hippocampus, and blood of male animals from the four inbred strains in response to CRS. Microarray analysis will be carried out on tissues obtained from animals exposed to: i) No stress, which will serve to determine the baseline gene expression ii.) 14 days of the CRS paradigm, which will inform us on the molecular markers of stress responsivity, and iii.) 10 days after the cessation of CRS to determine markers of post-stress responsivity. By employing microarray analyses of RNA isolated form two brain regions highly relevant to anxiety and stress and from blood, we will determine the common profiles of gene expression in these tissues during and after chronic stressor exposure. Specific Aim 3. Confirm the most promising candidate marker genes by quantitative real-time RT-PCR. This exploratory study will identify genes that are altered solely or concomitantly in specific brain regions or blood in response to chronic stress. These scenarios would allow us to identify candidate markers of chronic stress from the blood representing an accessible source of genetic material in humans.