Modulation of NO Synthase Gene Expression in CNS Neurons

描述（由申请人提供）：本项目的长期目标 应用是阐明一氧化氮的分子机制 合成酶I（NOS 1）基因表达在中枢和外周中受到调节， 神经系统对甲基苯丙胺诱导的、缺血的和创伤的反应 伤，分别。具体地说，我们希望识别和表征cis 调控人NOS 1基因转录的元件，并鉴定 因子和信号转导途径，其作用维斯这些元件， 诱导NOS 1来响应这些类型的损伤。 NOS 1是一个复杂的基因，至少有三个不同的启动子复合体 调节CNS中的转录。具体目标1建议确定CIS 对NOS 1启动子的CNS特异性表达重要的元件。 这些研究利用逆转录病毒，其表达EGFP报告基因， 通过单个人NOS 1启动子和细胞培养物的转录控制， 以及体内测定系统。具体目标2建议确定是否 个体人NOS 1启动子响应甲基苯丙胺而被激活 治疗、缺血和周围神经损伤。在这些 研究表明，表达不同NOS 1-lac Z融合基因的转基因小鼠品系 将进行三种神经元损伤模型中的每一种，然后 融合基因表达分析。具体目标3建议确定 这些融合基因是否在皮层神经元培养物中被激活， 产生兴奋性毒性的治疗和通过调节信号传导途径。 一氧化氮（NO）的表达与神经保护作用相关， 神经退行性疾病，并且取决于细胞环境，NO也充当神经退行性疾病的一部分。 强效神经毒素事实上，NO毒性是神经元细胞死亡的主要原因 中风期间。因此，了解所有流程至关重要， 有助于CNS中NOS的形成。通过关注转录 控制人NOS 1，该应用在以下方面具有巨大的潜在意义： 新药发现的背景下，并在提供更深刻的 了解CNS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.