Mouse-Strain specific Responses to NIcotine

小鼠品系对尼古丁的特异性反应

• 批准号: 6549069
• 负责人: SCOTT Warren ROGERS
• 金额: $ 32.59万
• 依托单位: UNIVERSITY OF UTAH
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-07-01 至 2007-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6549069
• 关键词: behavioral genetics; biological signal transduction; brain; drug addiction; endopeptidases; enzyme activity; gene targeting; genetic strain; genetic susceptibility; genetic transcription; immunologic techniques; laboratory mouse; microarray technology; nicotine; nicotinic receptors; oral administration; protein localization; protein structure function; proteolysis; receptor expression; smoking; tissue /cell culture; tobacco abuse; tumor necrosis factor alpha; video microscopy

DESCRIPTION (provided by applicant): Nicotine is the agent responsible for dependency to tobacco. But how does nicotine dependency, and the often-severe pathological consequences, develop in some, but not all, users? Nicotine exerts its effect(s) through specific ion-channel receptors termed nicotinic acetylcholine receptors (nAChRs). Because these receptors are the biological entry point through which nicotine exert its effects, understanding their expression and role in modulating cellular functions is imperative. However, the subunit composition and expression of native nAChRs is poorly defined as are the cellular mechanisms through which nicotine pre-conditions and modifies cellular responses. In this project we address these issues experimentally using genetically defined mouse strains with varying susceptibility to nicotine to define the distribution of key nAChR subunits in the brain. We will use microarray technology to define the impact of nicotine on transcription activation by the inflammatory cytokine, TNF alpha, and define proteolytic signaling cascades and responses that are altered by nicotine. Finally, we implement state-of-the-art gene-targeting and replacement methods to test directly the participation of key genes and/or pathways that are hypothesized to contribute to the genetic predisposition to nicotine as outlined below. Specific Aim 1. Strain-specific nAChR subunit distribution and expression. Hypothesis 1: The relative expression of nAChR subunit protein differs between inbred mouse strains. Hypothesis 2: Chronic nicotine (oral or smoking) alters region-specific nAChR subunit expression differently between inbred mouse strains. Specific Aim 2. Nicotine alters inflammatory transcriptional responses. Hypothesis: Strain-specific responses to nicotine pre-conditioning alter transcriptional response(s) induced by the pro-inflammatory cytokine, tumor necrosis alpha (TNFa). Specific Aim 3. Nicotine alters inflammatory post-transcriptional responses. Hypothesis: Strain specific responses to nicotine pre-conditioning modify protease activation and/or function induced by the proinflammatory cytokine, tumor necrosis alpha (TNF(x). Specific Aim 4. Strain specific zene-replacement of candidate genes. Hypothesis: Allelic replacement, through homologous recombination, of nAChR candidate gene polymorphisms from the C3H mouse into the C57BU6 will confer a novel phenotype on the recipient as defined in Specific Aims 1-3.

描述（由申请人提供）：尼古丁是负责烟草依赖的代理。 但是，尼古丁依赖性以及经常在某些但不是全部用户中发展的经常性病理后果如何？尼古丁通过称为烟碱乙酰胆碱受体（NACHRS）的特定离子通道受体发挥作用。由于这些受体是尼古丁发挥作用的生物学入口点，因此必须了解其调节细胞功能中的表达和作用。但是，天然NACHR的亚基组成和表达的定义很差，尼古丁预先提示并修饰细胞反应的细胞机制也是如此。在这个项目中，我们使用具有遗传定义的小鼠菌株在实验上解决这些问题，其对尼古丁的敏感性不同，以定义大脑中关键NACHR亚基的分布。我们将使用微阵列技术来定义尼古丁对炎症细胞因子，TNFα的影响，并定义尼古丁改变的蛋白水解信号级联和反应。最后，我们实施了最先进的基因靶向和替代方法，以直接测试关键基因和/或途径的参与，这些基因和/或途径被认为有助于促成 如下所述，对尼古丁的遗传易感性。 特定目标1。菌株特异性NACHR亚基分布和表达。假设1：NACHR亚基蛋白的相对表达在近交小鼠菌株之间有所不同。假设2：慢性尼古丁（口服或吸烟）在近交小鼠菌株之间改变了区域特异性的NACHR亚基表达。 具体目标2。尼古丁改变炎症转录反应。假设：促炎性细胞因子，肿瘤坏死α（TNFA）引起的尼古丁预先调节的菌株特异性反应。 特定目的3。尼古丁改变炎症后的转录后反应。假设：应变对尼古丁预调节的特异性反应修饰蛋白酶激活和/或由促炎细胞因子，肿瘤坏死α诱导的功能（TNF（TNF（x））。 特定目的4。候选基因的菌株特异性Zene替代。假设：通过同源重组，从C3H小鼠到C57BU6的NACHR候选基因多态性的等位基因替代将授予特定目标1-3中定义的接受者的新表型。