MODULATION OF NO SYNTHASE GENE EXPRESSION IN CNS NEURONS

这项建议的目的是阐明分子机制。 人神经元型一氧化氮合酶的转录调控 (NOS1)基因。这将通过识别基因 允许人类NOS1基因的CNS表达的元件； 决定如何通过这些来实现转录控制 元素；以及创造以转基因小鼠为基础的模型来研究人类 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 mRNAs在 CNS由不同的启动子组成。目标1建议检查结构 以及这些5‘末端外显子的分布。目标2建议确定 顺式作用元件对于每个启动子的功能和 确定在发育、再生和/或 受创伤的神经元与这些元素相互作用。目标3建议 确定4.3kb的人NOS1启动子复合体 对足够的信息进行编码，以指导适当监管 LawZ在嗅觉感受器神经元中的表达 构建了转基因小鼠品系。目标4建议生成线条 NOS1基因大片段转基因小鼠的Z表达规律 以确定NOS1基因的顺式作用元件是否可以 根据Z定律确定基因表达的时空模式 报道称，内源性NOS1在整个细胞内的表达 整个中枢神经系统在发育过程中和对神经创伤的反应中。 这项研究之所以重要，有几个原因。第一，建立 在中枢神经系统中支撑NOS1转录的基本机制是 标志着朝着发展药理学能力迈出的重要一步 调节NOS1基因的表达，进而调节中枢神经系统中NOS1蛋白的表达。 大量数据表明生理和病理生理学 一氧化氮合酶在中枢神经系统中的作用，提示临床治疗可能 从这样的能力中脱颖而出。第二，NOS1的复杂格局 基因表达，涉及地区特有的产前和出生后 改变，表明与神经元普遍相关的原则 发展可能源于对NOS1的研究 抄写。第三，NOS1在中枢神经系统中的转录受 两个紧密相连但又可以分开的推动者。澄清： 这种新的基因组结构调节NOS1的机制， 基因表达应该扩大我们对基因表达的普遍理解 转录调控是在中枢神经系统实现的。

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.