NFKB FUNCTION IN REGULATING NEURONAL GENE EXPRESSION

描述(改编自申请者摘要)：神经元凋亡和 变性是神经系统多种严重疾病的常见病症。 包括创伤性脑损伤、局灶性脑缺血、帕金森氏症和 阿尔茨海默病。除了控制基本的生理 在这一过程中，核因子-kB转录因子家族已经成为主要的 许多系统中细胞生死的调节者。自从他们最近的发现 在神经系统中，促凋亡和抗凋亡的作用都是 归因于核因子-kB。我们的建议将解决不同的可能性 神经刺激的机制可能导致时间上的差异 活化神经元核因子-kB的进程、程度和亚基组成。这些 反过来，差异可以解释不同的生理功能 神经元NFkB在不同的环境中，并可能导致 治疗调节核因子-kB激活的策略。我们已经选择了 海马体作为我们研究神经元核因子-kB的模型系统，因为它是一个 明确界定的生理区域(学习和记忆)以及 病理性(中风、阿尔茨海默病)神经元功能。我们的研究将 使用成年小鼠和新生小鼠的海马区组织以及几个 转基因和基因敲除品系。我们将确定核因子-kB家族成员 存在于海马神经元中，并定义了导致 它们的激活。我们将调查具有代表性的神经递质， 神经营养因子和细胞黏附分子的激活能力 使用电迁移率移位分析和kB报告构建的神经元核因子-kB。 此外，我们将使用荧光标记的NF-kB亚单位来检测 核因子-kB的亚细胞定位及其转位能力 神经突起的核遵循不同的刺激参数。 我们将检查突触活动水平是否在正常生理范围内 Range功能调节NFkB，或者如果仅在 对应激刺激的反应，NFkB的激活模式能否编码 关于刺激的生理性质和毒性性质的信息？电气 刺激将用于评估NFkB的激活模式和 依赖于kB的基因表达对精确变异刺激的反应 强度、持续时间和频率。我们的调查将做出新的贡献 通过具体研究核因子-kB家族成员在 生理或病理生理学对神经元基因表达的调节 刺激物。 作为一名专门研究神经科学的医学博士，我希望自己的职业是 一名研究转录调控和 中枢神经系统疾病的发病机制。

DESCRIPTION (Adapted From The Applicant's Abstract): Neuronal apopotosis and degeneration is common to a wide range of severe diseases in the nervous system including traumatic brain injury, focal ischemia, Parkinsons's disease and Alzheimers disease. In addition to the control of basic physiological processes, the NF-kB family of transcription factors have emerged as major regulators of cell life and death in many systems. Since their recent discovery in,the nervous system, both pro- and anti-apoptotic actions have been attributed to NF-kB. Our proposal will address the possibility that different mechanisms of neuronal stimulation may result in differences in the time course, degree, and subunit composition of activated neuronal NF-kB. These differences, in turn, could explain contrasting physiologic functions of neuronal NFkB in disparate settings and might lead to the development of strategies to therapeutically regulate NF-kB activation. We have selected the hippocampus as our model system for studying neuronal NF-kB because it is a well-defined area of both physiological (learning and memory) as well as pathological (stroke, Alzheimers disease) neuronal function. Our studies will use hippocampal tissue from both adult mice and neonates as well as several transgenic and knockout lines. We will identify the NF-kB family members present in hippocampal neurons and define the types of stimulation which lead to their activation. We will investigate representative neurotransmitters, neurotrophins, and cell adhesion molecules for their ability to activate neuronal NF-kB using electromobility shift assays and a kB-reporter construct. In addition, we will use a fluorescently labeled NF-kB subunit to examine the subcellular localization of NF-kB and determine its ability to translocate to the nucleus from neuronal processes following different stimulation parameters. We will examine if levels of synaptic activity within the normal physiological range functionally regulate NFkB, or if significant activation occurs only in response to stressful stimuli, Can the pattern of NFkB activation encode information on the physiological versus toxic nature of a stimulus? Electrical stimulation will be used to assess the pattern of NFkB activation and kBdependent gene expression in response to stimuli of precisely varied intensity, duration, and frequency. Our investigations will contribute new knowledge by specifically examining the roles of NF-kB family members in the regulation of neuronal gene expression by physiological or pathophysiological stimuli. As an M.D.- Ph.D. specializing in the neurosciences, I hope to have a career as an independent investigator studying transcriptional regulation and the pathogenesis of central nervous system disease.