NFKB FUNCTION IN REGULATING NEURONAL GENE EXPRESSION

描述（改编自申请人的摘要）：神经元凋亡和 变性是神经系统中多种严重疾病的常见症状 包括创伤性脑损伤、局灶性缺血、帕金森病和 老年痴呆症。除了控制基本的生理 在这些过程中，NF-κ B家族的转录因子已经成为主要的 在许多系统中调节细胞的生死。自从他们最近发现 在神经系统中，促细胞凋亡和抗细胞凋亡作用都已被证实。 归因于NF-kB。我们的建议将解决不同的可能性， 神经元刺激的机制可能导致时间的差异 神经元NF-κ B活化的过程、程度和亚基组成。这些 差异，反过来，可以解释不同的生理功能， 神经元NFkB在不同的设置，并可能导致发展 治疗性调节NF-κ B活化的策略。我们选择了 海马作为我们研究神经元NF-kB的模型系统，因为它是一个 生理（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组织以及几种 转基因和敲除品系。我们将确定NF-κ B家族成员 存在于海马神经元中，并定义了导致 他们的激活。我们将研究代表性的神经递质， 神经营养因子和细胞粘附分子，因为它们能够激活 神经元NF-κ B使用电迁移率变动分析和κ B-报告构建体。 此外，我们将使用荧光标记的NF-kB亚基来检查 NF-kB的亚细胞定位并确定其易位至 在不同的刺激参数下，神经元过程的核。 我们将检查正常生理条件下的突触活动水平， 范围在功能上调节NFkB，或者如果仅在 应激反应，NFkB激活模式是否编码 刺激物的生理性质与毒性性质的信息？电 刺激将用于评估NFkB激活的模式， KB依赖性基因表达对精确变化的刺激的响应 强度、持续时间和频率。我们的调查将为 通过专门研究NF-kB家族成员在肿瘤发生中的作用， 调节神经元基因表达的生理或病理生理 刺激。 作为医学博士-博士我的专业是神经科学，我希望有一个职业生涯， 一个独立的研究人员研究转录调控和 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.