MODULATION OF NO SYNTHASE GENE EXPRESSION IN CNS NEURONS

本研究的目的是阐明其分子机制 人神经元型一氧化氮合酶的转录调控 （NOS1）基因。 这将通过识别遗传 允许人NOS1基因CNS表达的元件; 确定转录控制是如何实现的， 元素;以及创建基于转基因小鼠的模型来研究人类 NOS1启动子在发育、再生和创伤中的作用 神经元 具有不同5 '末端外显子的NOS 1 mRNA在细胞中转录， CNS通过单独的启动子。 目标1建议审查结构 以及这些5 '端外显子的分布。 目标2：确定 每个启动子的功能所必需的顺式作用元件， 确定在发育、再生和/或再生过程中是否存在因素， 受到创伤的神经元与这些元素相互作用。 目标3建议， 确定4.3kb人NOS 1启动子复合物是否 编码足够的信息，以指导适当的监管 表达的法律Z嗅觉受体神经元，使用以前 构建转基因小鼠品系。Aim 4建议生成线条 law z-表达转基因小鼠的NOS 1较长片段 基因，以确定NOS 1基因的顺式作用元件是否可以 赋予基因表达的空间和时间模式的法律Z 报告，反映整个内源性NOS 1的表达， 整个中枢神经系统在发展过程中和神经创伤的反应。 这项研究之所以重要，有几个原因。 第一，建立 支持CNS中NOS 1转录的基本机制将 标志着朝着发展药理学能力迈出了重要的一步， 调节NOS 1 mRNA，从而调节CNS中的NOS 1蛋白。 大量的数据表明， NOS在CNS中的作用，表明临床治疗可能 从这样的能力中走出来。 第二，NOS1的复杂模式 基因表达，涉及区域特异性产前和产后 改变，表明神经元的一般相关性原则， 发展可能来自NOS1的研究 转录。 第三，NOS 1在CNS中的转录受以下因素控制： 两个紧密相连但可分离的启动子。 阐明 这种新的基因组结构调节NOS 1的机制， 基因表达应该会扩大我们对 在CNS中实现转录控制。

The object of this proposal is to elucidate the molecule mechanisms regulation transcription of the human neuronal nitric oxide synthase (NOS1) gene. This will be accomplished by identifying genetic elements that permit CNS expression of the human NOS1 gene; determining how transcriptional control is achieved vis a vis these elements; and creating transgenic mouse-based models to study human NOS1 promoter function in developing, regeneration, and traumatized neurons. NOS 1 mRNAs with different 5' terminal exons are transcribed in the CNS by separate promoters. Aim 1 proposes to examine the structure and distribution of these 5' terminal exons. Aim 2 proposes to identify cis acting elements essential for functioning of each promoter and to determine whether factors present in developing, regenerating, and/or traumatized neurons interact with these elements. Aim 3 proposes to determine whether the 4.3 kb human NOS1 promoter complex encodes sufficient information to direct appropriately regulated expression of law Z in olfactory receptor neurons, using previously constructed lines of transgenic mice. Aim 4 proposes to generate lines of law z-expression transgenic mice with longer fragments of the NOS1 gene, to determine whether cis-acting elements of the NOS1 gene can confer a spatial and temporal pattern of gene expression on the law Z reporter that reflects expression of endogenous NOS1 throughout the entire CNS during development and in response to neurotrauma. This research is important for several reasons. First, establishing the basic mechanisms that underpin NOS1 transcription in the CNS would mark an essential step toward developing a pharmacologic ability to regulate NOS1 mRNA, and consequently NOS 1 protein in the CNS. An avalanche of data point out physiological and pathophysiological roles of NOS in the CNS, suggesting that clinical treatments may emerge from such an ability. Second, the complex pattern of NOS1 gene expression, involving regionally specific prenatal and postnatal alterations, indicates that principles of general relevance to neuronal development are likely to be derived from studies of NOS1 transcription. Third, transcription of NOS1 in the CNS is controlled by two closely linked but separable promoters. Elucidation of the mechanisms by which this novel genomic structure regulates NOS1, gene expression should expand our general appreciation of how transcriptional control is achieved in the CNS.