Quantitative Genetic Study of Seizures

癫痫发作的定量遗传学研究

• 批准号: 7340461
• 负责人: THOMAS N FERRARO
• 金额: $ 47.99万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-09-01 至 2009-09-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7340461
• 关键词: Accounting; Address; Applications Grants; Bacterial Artificial Chromosomes; Candidate Disease Gene; Chromosomes, Human, Pair 1; Chromosomes, Human, Pair 15; Chromosomes, Human, Pair 5; Classification; Complex; Congenic Strain; Data; Dissection; Electroconvulsive Shock; Electrophysiology (science); Epilepsy; Evaluation; Event; Exhibits; Facility Construction Funding Category; Foundations; Funding; Future; Generalized Epilepsy; Genes; Genetic; Genetic Predisposition to Disease; Genetic Variation; Genetically Engineered Mouse; Goals; Homologous Gene; Human; In Vitro; Inbred Mouse; Inbred Strains Mice; Injection of therapeutic agent; Ion Channel Protein; Kainic Acid; Knock-in Mouse; Lead; Location; Maps; Methods; Modeling; Mouse Strains; Mus; Nature; Pentylenetetrazole; Phase; Positioning Attribute; Potassium Channel; Predisposition; Property; Quantitative Genetics; Quantitative Trait Loci; Regulation; Request for Applications; Research; Research Design; Resistance; Screening procedure; Seizures; Staging; Surveys; Susceptibility Gene; Testing; Transgenic Organisms; Translating; Variant; Work; base; behavior test; concept; gene cloning; homologous recombination; in vivo; insight; neuronal excitability; programs; prototype; success

This application requesting 4 years of funding to continue a comprehensive program in the study of genetic susceptibility to seizures. The long term goal of this work is to identify seizure susceptibility genes in inbred mice and use them as the basis for focused studies in humans with common forms of epilepsy. In the initial stages of this project, we used quantitative trait locus (QTL) mapping to study seizure susceptibility in 2 inbred mouse strains: B6, which is resistant to experimental seizures, and D2, which is susceptible. Results of this work identified a locus of major effect, Szs1 (chr.1), and several other loci of moderate effect including Szs11 (chr. 5) and Szs13 (chr. 15). We hypothesize that the genes underlying these loci are fundamentally important in the regulation of neuronal excitability since they were detected consistently in diverse screening paradigms using kainic acid, pentylenetetrazol and electroshock. Thus, over the past funding period, we have focused on characterizing each of these loci further, primarily through a congenic strain-candidate gene strategy. Results have led to nomination of a gene for Szs1 and confirmation of Szs11 and Szs13 in congenic strains. The putative Szs1 gene is Kcnj10 which encodes an inward-rectifying potassium channel and plans to generate formal proof that it is Szs1 comprise the first aim of this proposal. Studies will involve creation of transgenic and knock-in mice and will utilize in vivo and in vitro electrophysiology as well as behavioral tests of seizure susceptibility. Aim 2 involves continued refinement of Szs11 and Szs13 using the prototype strategy we developed for Szs1. Thus, based on completion of congenic strains begun during the last funding period and systematic creation of interval specific congenic strains, we plan to reduce the critical intervals to 3 cM and then begin candidate gene analysis. The third aim involves initiation of new QTL studies designed to exploit the phenotypic diversity exhibited by other inbred mouse strains for identifying seizure susceptibility loci. Overall, we hypothesize that genetic variation associated with seizure susceptibility in mice is related to that which underlies common human epilepsies; and further, that by identifying key loci in mice greater insight will be afforded into mechanisms of seizures and their abolition in humans. Recent data from our lab documenting a significant association between variants of KCNJ10 and idiopathic generalized epilepsy support this concept and justify the continued use of QTL mapping for identifying seizure susceptibility loci in mice.

该申请要求4年的资金继续一项全面的计划，以研究遗传易感性。这项工作的长期目标是识别近交小鼠中的癫痫敏感性基因，并将其用作具有常见癫痫病的人类的集中研究的基础。在该项目的初始阶段，我们使用定量性状基因座（QTL）映射来研究2种近交小鼠菌株中的癫痫敏感性：B6，对实验性癫痫发作具有抵抗力，而D2具有易感性。这项工作的结果确定了一个主要影响的基因座，SZS1（CHR.1）和其他几个中等效果的基因座，包括SZS1111（Chr。5）和SZS13（Chr。15）。我们假设这些基因座的基因从根本上是 对于调节神经元兴奋性很重要，因为使用Kainic Acid，五乙醇唑和电摇头在不同的筛选范式中始终检测到它们。因此，在过去的资金期间，我们专注于进一步表征这些基因座，主要是通过先天性应变基因策略。结果导致提名了SZS1基因，并确认了先天性菌株中SZS11和SZS13的确认。推定的SZS1基因是KCNJ10，它编码了一个内向的矫正钾渠道，并计划产生正式证明它是SZS1构成该提案的第一个目的。研究将涉及创建转基因和敲门小鼠，并将在体内和体外电生理学以及癫痫敏感性的行为测试中使用。 AIM 2涉及使用我们为SZS1开发的原型策略对SZS11和SZS13的持续改进。因此，基于在最后一个资金期间开始的先天性菌株的完成，并系统地创建了间隔特定的先天性菌株，我们计划将关键间隔减少到3 cm，然后开始候选基因分析。第三个目的涉及启动新的QTL研究，旨在利用其他近交小鼠菌株表现出的表型多样性，以识别癫痫发作易感性基因座。总体而言，我们假设与小鼠的癫痫敏感性相关的遗传变异与人类常见癫痫的基础相关。此外，通过识别小鼠的关键基因座，将提供更大的洞察力，以癫痫发作的机制及其在人类中的废除。来自我们实验室的最新数据记录了KCNJ10变体与特发性广泛性癫痫之间的显着关联，并证明了继续使用QTL映射以识别小鼠中的癫痫发作位点是合理的。