Constructing a Conditional Kalirin Null Mouse

构建有条件的 Kalirin 空小鼠

• 批准号: 6954668
• 负责人: RICHARD E MAINS
• 金额: $ 14.5万
• 依托单位: UNIVERSITY OF CONNECTICUT SCH OF MED/DNT
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-09-30 至 2006-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6954668
• 关键词: alleles; confocal scanning microscopy; developmental neurobiology; gene targeting; genetically modified animals; guanine nucleotide binding protein; hippocampus; laboratory mouse; mutant; nervous system; neural plasticity; nucleus accumbens; polymerase chain reaction; protein isoforms; protein quantitation /detection; protein structure function

DESCRIPTION (provided by applicant): Kalirin is a large, multidomain Rho GDP/GTP exchange factor (GEF) of the Dbl family found almost exclusively in the nervous system. Based on genetic studies in invertebrates and analysis of genes involved in X-linked mental retardation, Kalirin is expected to play a role in axonal pathfinding and synapse formation. Our studies in rat hippocampal, cortical and sympathetic neurons showed that Kalirin plays a role in axon initiation and outgrowth, dendritic growth, and spine formation and maintenance. Kalirin has two GEF domains with specificity for different Rho family GTPases and a potential kinase domain. In addition to these catalytic domains, it has interactor domains for a variety of proteins. Kalirin expression in the nucleus accumbens and striatum is stimulated by chronic administration of cocaine, with no change in cortical Kalirin levels; Kalirin is also increased following electroconvulsive shock stimulation. The exploratory studies proposed are based on our observations that antisense-mediated elimination of Kalirin expression in organotypic slices and dissociated hippocampal neurons leads first to a reduction in linear spine density followed by simplification of the dendritic tree. Aim 1 is to generate mutant mice that will facilitate studies of neuronal development and plasticity in multiple systems. In order to avoid embryonic or early postnatal lethality, and to allow generation of tissue-specific and developmentally regulated elimination of expression, two mouse lines will be created as floxed alleles. Both will be provided to all qualified investigators without restrictions. One allele will produce the Kalirin-7 null. Kalirin-7 is the most prevalent isoform in the adult brain, with a single Rho GEF domain, terminating with a unique PDZ binding motif. Kalirin-7 appears late in development and is localized to dendritic spines. The second allele will produce the total Kalidn knockout. Both will be bred with available tissue-specific and drug-inducible Cre recombinase mice. Aim 2 will examine the biochemical, histological, and developmental consequences of tissue-specific Kalirin conditional knockouts.

描述（由申请人提供）： Kalirin是Dbl家族的大的多结构域Rho GDP/GTP交换因子（GEF），几乎仅在神经系统中发现。基于对无脊椎动物的遗传研究和对X连锁精神发育迟滞相关基因的分析，Kalirin有望在轴突寻路和突触形成中发挥作用。我们在大鼠海马、皮质和交感神经元中的研究表明，Kalirin在轴突起始和生长、树突生长以及棘形成和维持中起作用。Kalirin具有两个对不同Rho家族GTP酶具有特异性的GEF结构域和一个潜在的激酶结构域。除了这些催化结构域之外，它还具有多种蛋白质的相互作用结构域。Kalirin的表达在丘脑核和纹状体的刺激慢性管理的可卡因，皮质Kalirin水平没有变化; Kalirin电休克刺激后也增加。提出的探索性研究是基于我们的观察，反义介导的消除卡利林表达的器官型切片和解离的海马神经元导致首先减少线性棘密度，然后简化的树突树。目的1是产生突变小鼠，这将有助于在多个系统中研究神经元发育和可塑性。为了避免胚胎或出生后早期致死，并允许产生组织特异性和发育调节的表达消除，将创建两个小鼠品系作为floxed等位基因。这两份报告将无限制地提供给所有合格的研究者。一个等位基因将产生Kalirin-7无效。Kalirin-7是成人脑中最普遍的同种型，具有单个Rho GEF结构域，以独特的PDZ结合基序终止。Kalirin-7在发育后期出现，并定位于树突棘。第二个等位基因将产生总Kalidn敲除。两者都将与可用的组织特异性和药物诱导型Cre重组酶小鼠一起繁殖。目的2将研究组织特异性Kalirin条件性敲除的生物化学、组织学和发育后果。