Modulation of NO Synthase Gene Expression in CNS Neurons

描述(由申请人提供)：这项计划的长期目标 应用是阐明一氧化氮的分子机制 合成酶I(NOS1)基因的表达在中枢和外周受到调节 神经系统对甲基苯丙胺诱导的、缺血的和创伤的反应 伤情分别为。具体地说，我们希望识别和描述CI 调控人类NOS1基因转录的元件，并鉴定 作用于这些元件的因子和信号转导途径 诱导NOS1对这些类型的伤害做出反应。 Nos1是一个复杂的基因，至少有三个不同的启动子复合体。 调节中枢神经系统中的转录。具体目标1建议确定独联体 对于NOS1启动子的CNS特异性表达很重要的元件。 这些研究利用逆转录病毒表达EGFP报告基因 单个人NOS1启动子和细胞培养的转录调控 以及体内检测系统。具体目标2提议确定是否 甲基苯丙胺激活单个人类NOS1启动子 治疗、缺血和周围神经损伤。在这些 表达不同NOS1-lac Z融合基因转基因小鼠品系的研究 会分别受到三种神经元损伤模型的影响，其次是 融合基因表达分析。具体目标3建议确定 这些融合基因在皮质神经元培养中是否被激活 产生兴奋性毒性的治疗和信号通路的调节。 一氧化氮(NO)表达与神经保护的关系 神经退行性疾病，根据细胞环境的不同，一氧化氮也起着 强效神经毒素。事实上，没有毒性是神经细胞死亡的主要原因。 在中风期间。因此，了解符合以下条件的所有流程至关重要 促进中枢神经系统一氧化氮合酶的形成。通过专注于转录 人一氧化氮合酶1的调控，这一应用具有很大的潜在意义 新药发现的背景和在提供更深刻的 了解中枢神经系统中基因表达的可塑性。

DESCRIPTION (provided by applicant): The long term objective of this application is to elucidate the molecular mechanisms by which nitric oxide synthase I (NOS1) gene expression is modulated in the central and peripheral nervous systems in response to methamphetamine-induced, ischemic, and traumatic injury, respectively. Specifically, we hope to identify and characterize cis elements that regulate transcription of the human NOS1 gene, and to identify factors and signal transduction pathways that act vis a vis these elements to induce NOS1 in response to these types of injuries. NOS1 is a complex gene with at least three distinct promoter complexes regulating transcription in the CNS. Specific aim 1 proposes to identify cis elements that are important for CNS-specific expression of the NOS1 promoters. These studies utilize retroviruses that express an EGFP reporter under transcriptional control by individual human NOS1 promoters and cell culture as well as in vivo assay systems. Specific aim 2 proposes to determine whether individual human NOS1 promoters are activated in response to methamphetamine treatment, ischemia, and peripheral nerve injury, respectively. In these studies, lines of transgenic mice expressing different NOS1-lac Z fusion genes will be subjected to the each of the three neuronal injury models, followed by analysis of fusion gene expression. Specific aim 3 proposes to determine whether these fusion genes are activated in cortical neuronal cultures by treatments producing excitotoxicity and by modulation of signaling pathways. Expression of nitric oxide (NO) is correlated with neuroprotection in neurodegenerative disease and, depending on cellular context, NO also acts as a potent neurotoxin. Indeed, NO toxicity is a major cause of neuronal cell death during stroke. Consequently, it is critical to understand all processes that contribute to the formation of NOS in the CNS. By focusing on transcriptional control of human NOS 1, this application has great potentia1 significance in the context of new drug discovery and in providing a more profound understanding of the plasticity of gene expression in the CNS.