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A conditional, tissue specific 5-HT1B knockout mouse

条件性、组织特异性 5-HT1B 基因敲除小鼠

基本信息

批准号：
8701407
负责人：
John F Neumaier
金额：
$ 19.31万
依托单位：
UNIVERSITY OF WASHINGTON
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-07-15 至 2015-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8701407
关键词：
Ablation Adult Aggressive behavior Agonist Animal Model Anxiety Autoreceptors Behavior Behavior Control Behavioral Biological Assay Brain Brain region Breeding Cells Complex Corpus striatum structure Development Disease Drug Addiction Drug usage Eating Eating Behavior Eating Disorders Emotional Emotional disorder Event Financial compensation Fright Future Gene Deletion Gene Targeting Genes Genetic Recombination Glutamates Goals Headache Human Impulsive Behavior Knock-out Knockout Mice Measures Mediating Mental Depression Mental disorders Methods Migraine Modeling Mus Neurobiology Neurons Neurotransmitters Obsessive compulsive behavior Obsessive-Compulsive Disorder Pathway interactions Pattern Phenotype Population Presynaptic Terminals Problem Solving Regulation Research Personnel Respiration Role Serotonin Serotonin Receptor 5-HT1B Site Sorting - Cell Movement Specificity Stress Sudden infant death syndrome Tamoxifen Testing Tissues Transgenic Mice Transgenic Organisms Virus addiction cell type cholinergic neuron conditioned fear design dorsal raphe nucleus flexibility homologous recombination innovation knockout gene neural circuit neurotransmitter release novel overexpression public health relevance receptor receptor expression response serotonin transporter tool

项目摘要

DESCRIPTION (provided by applicant): 5-HT1B receptors are expressed in diverse neuron types throughout the brain where they act as inhibitory receptors on presynaptic terminals. When expressed by serotonergic neurons they are autoreceptors whereas in other types of neurons they act as heteroreceptors. It has been difficult to sort out the role of 5-HT1B autoreceptors and heteroreceptors because they are intermixed in most brain regions, even though they are regulating the release of different neurotransmitters. Furthermore, constitutive knockout of 5-HT1B receptors have a complex phenotype that may reflect developmental compensations predominantly, instead of informing about the role of these receptors in adult brain function. Therefore, two types of conditional expression are needed as tools to investigate the contribution of 5-HT1B receptors in different neurons to complex emotional behavior: temporal and phenotype specificity of gene knockout. This proposal intends to solve this problem by creating a new transgenic mouse that will allow conditional and cell type-specific deletion or protection of 5-HT1B receptor expression using available Cre and Flp driver lines. The proposed transgenic mouse is innovative for several reasons. 1. Conditional, cell type-specific expression of Cre will delete the gene in the targeted neurons. 2. Conditional, cell type-specific expression of Flp will excise a loxP site thereby selectively protecting the 5-HT1B gene in those neurons while remaining 5-HT1B receptors elsewhere can be subsequently knocked out by Cre. 3. The targeting construct is designed to minimize the chances of baseline reduction in 5-HT1B expression prior to deletion (i.e. hypomorphism). 4. This strategy can be applied to many different situations to target (or preserve) 5-HT1B receptors with any available Cre and Flp driver lines. For this revised R21 proposal we will focus our characterization on the conditional knockout of 5-HT1B autoreceptors just in serotonergic neurons. In Aim 1 we will construct the targeting construct, express it via homologous recombination, derive transgenic mice, and characterize the behavioral phenotype of the mice. In Aim 2 we will examine the impact of the selective knockout of 5-HT1B autoreceptors (in serotonin neurons) on serotonin transporter function and conditioned fear, an animal model relevant to a number of psychiatric disorders. This will allow us to probe our hypothesis that 5-HT1B autoreceptors are the primary site of 5-HT1B-mediated reductions in fear behavior definitively. In the future it will be possible to examine the site of action of 5-HT1B drugs by using a wide range of other Cre and Flp driver lines to investigate other important neurobiological problems including models of drug addiction, regulation of eating behaviors, and control of respiration in animal models of Sudden Infant Death Syndrome.

描述（由申请人提供）：5-HT 1B受体在整个大脑的不同神经元类型中表达，在那里它们作为突触前末梢上的抑制性受体。当由多巴胺能神经元表达时，它们是自身受体，而在其他类型的神经元中，它们充当异源受体。很难区分5-HT 1B自身受体和异源受体的作用，因为它们在大多数大脑区域混合，即使它们调节不同神经递质的释放。此外，5-HT 1B受体的组成性敲除具有复杂的表型，其可能主要反映发育补偿，而不是告知这些受体在成人脑功能中的作用。因此，需要两种类型的条件表达作为工具来研究不同神经元中的5-HT 1B受体对复杂情绪行为的贡献：基因敲除的时间特异性和表型特异性。该提案旨在通过创建新的转基因小鼠来解决这个问题，该转基因小鼠将允许使用可用的Cre和Flp驱动系对5-HT 1B受体表达进行条件性和细胞类型特异性缺失或保护。提出的转基因小鼠是创新的几个原因。1.有条件的，细胞类型特异性表达的Cre将删除靶向神经元中的基因。2. Flp的有条件的细胞类型特异性表达将切除loxP位点，从而选择性地保护那些神经元中的5-HT 1B基因，而其他地方的剩余5-HT 1B受体随后可以被Cre敲除。3.靶向构建体被设计为使5-HT 1B表达在缺失之前基线降低的机会最小化（即，亚型）。4.这种策略可以应用于许多不同的情况，以靶向（或保存）5-HT 1B受体与任何可用的Cre和Flp驱动线。对于这个修订的R21建议，我们将集中我们的特性的条件敲除5-HT 1B自身受体只是在肾上腺素能神经元。在目标1中，我们将构建靶向构建体，通过同源重组表达它，获得转基因小鼠，并表征小鼠的行为表型。在目标2中，我们将研究选择性敲除5-HT 1B自身受体（5-羟色胺神经元）对5-羟色胺转运体功能和条件性恐惧的影响，这是一种与许多精神疾病相关的动物模型。这将使我们能够探索我们的假设，即5-HT 1B自身受体是5-HT 1B介导的恐惧行为减少的主要部位。在未来，通过使用广泛的其他Cre和Flp驱动线来检查5-HT 1B药物的作用位点，以研究其他重要的神经生物学问题，包括药物成瘾模型、进食行为的调节和婴儿猝死综合征动物模型中的呼吸控制。