Novel Brain-Selective Conditional Knockout of the Norepinephrine Transporter

新型脑选择性条件性敲除去甲肾上腺素转运蛋白

• 批准号: 8701598
• 负责人: MAUREEN K HAHN
• 金额: $ 23.5万
• 依托单位: VANDERBILT UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-03-04 至 2016-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8701598
• 关键词: Active Biological Transport; Alleles; Anxiety; Behavior; Blood Pressure; Brain; Brain Diseases; Cardiovascular Diseases; Cardiovascular Physiology; Cardiovascular system; Catecholamines; Comorbidity; Disease; Dissection; Exhibits; Functional disorder; Genes; Genetic; Genetic Recombination; Goals; Heart; Heart Rate; Homeostasis; Human; Incidence; Knock-in Mouse; Knock-out; Knockout Mice; Measures; Mediator of activation protein; Medicine; Mental Depression; Mental disorders; Modeling; Moods; Mus; Mutation; Neurons; Neurotransmitters; Norepinephrine; Palpitations; Pathway interactions; Patients; Performance; Peripheral; Pharmaceutical Preparations; Phenotype; Play; Reporter; Reporter Genes; Reporting; Role; Rosa; Signal Transduction; Stress; Sympathetic Nervous System; Synapses; Tachycardia; Tail Suspension; Testing; Tissues; Tomatoes; Translating; Work; genetic risk factor; heart innervation; mind control; mouse model; neurobehavioral; noradrenaline transporter; noradrenergic; novel; offspring; presynaptic; promoter; public health relevance; tool

DESCRIPTION (provided by applicant): The aim of this proposal is to develop novel mouse models that produce conditional knockout of the norepinephrine transporter (NET) gene. The overarching goal of this work is to elucidate the role of NET in central versus peripheral sympathetic nervous system (CNS vs. SNS) noradrenergic neurons to influence psychiatric phenotypes, and inspire new treatments for such disorders. Increasing evidence indicates significant comorbidities between brain and cardiovascular disorders. For example, patients with anxiety complain of heart palpitations and patients with tachycardia disorders have a higher incidence of anxiety. How these comorbidities derive from shared genetic risk factors is poorly understood. One shared component of circuits controlling mood and cardiovascular function is the neurotransmitter, norepinephrine (NE). NE released at synapses in the brain and heart is inactivated through active transport into terminals by the presynaptically- localized NET, making NET a critical mediator of NE inactivation and presynaptic catecholamine homeostasis. We have demonstrated that mice with a constitutive NET knockout or knock-in of a non-functional NET mutation, A457P, exhibit both tachycardia and anxiety. We have now generated the first mouse with a "floxed" NET allele to allow selective NET knockout to dissect CNS vs. SNS contributions to psychiatric disease. There has been little study to date of promoters that can drive expression in noradrenergic neurons selectively in the CNS versus SNS. This proposal will determine the ability of CRE driver lines to selectively knock out NET in central versus peripheral noradrenergic neurons. Aim I will cross CRE lines that reportedly drive recombination in either brain or SNS NE neurons with Rosa reporter mice and NET floxed mice. We will determine reporter expression, and NET expression and activity in the brain and SNS. Aim II will determine if anxiety/depression- related behaviors observed with constitutive manipulation of NET are driven by brain versus SNS NET. These studies introduce the novel scientific idea to use genetic dissection of brain versus SNS noradrenergic function, and provide a novel characterization of tools to accomplish this. These models will translate to the study of many other genes with tissue-specific expression in noradrenergic neurons. This work will allow us to more precisely understand the role of brain noradrenergic signaling in mental disorders.

描述（由申请人提供）：该建议的目的是开发新型的小鼠模型，以产生去甲肾上腺素转运蛋白（NET）基因的条件敲除。这项工作的总体目标是阐明网络在中央和周围交感神经系统（CNS vs. SNS）中脱肾上腺素能神经元的作用，以影响精神病表型，并激发对此类疾病的新疗法。越来越多的证据表明大脑和心血管疾病之间的合并症显着。例如，焦虑症患者对心pal和心动过速疾病患者的焦虑发生率更高。这些合并症是如何从共同的遗传危险因素中得出的。控制情绪和心血管功能的电路的共享组成部分是神经递质，去甲肾上腺素（NE）。通过突触前局部网络使大脑和心脏突触中的NE释放到大脑和心脏的释放，使网络成为NE灭活和突触前儿茶酚胺稳态的关键介体。我们已经证明，非功能性净突变A457p的净敲除或敲除的小鼠表现出心动过速和焦虑。现在，我们已经产生了第一只带有“ Floxed”净等位基因的小鼠，以允许选择性净敲除剖析CNS与SNS对精神病的贡献。迄今为止，启动子的研究很少，可以在CNS与SNS中有选择地推动在去甲肾上腺素能神经元中的表达。该提案将确定CRE驱动线在中心与外周去甲肾上腺素能神经元中有选择性地淘汰网的能力。目的，我将越过CRE线，据报道，用Rosa报告的小鼠和净floxed小鼠在大脑或SNS NE神经元中驱动重组。我们将确定记者的表达，以及大脑和SN中的净表达和活性。 AIM II将确定通过NET的构成操纵观察到的焦虑/抑郁症相关行为是否由大脑与SNS网络驱动。这些研究介绍了使用大脑与SNS去甲肾上腺素能功能的遗传解剖的新型科学思想，并提供了实现这一目标的工具的新颖特征。这些模型将转化为对多肾上腺素能神经元中组织特异性表达的许多其他基因的研究。这项工作将使我们更准确地了解脑甲肾上腺素能信号在精神障碍中的作用。