Mammalian genetic analysis of embryonic neural overgrow*

胚胎神经过度生长的哺乳动物遗传分析*

• 批准号: 6708073
• 负责人: KAREN J ARTZT
• 金额: $ 36.88万
• 依托单位: UNIVERSITY OF TEXAS AT AUSTIN
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-03-15 至 2007-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6708073
• 关键词: animal genetic material tag; artificial chromosomes; athymic mouse; developmental genetics; developmental neurobiology; disease /disorder model; gene mutation; genetic mapping; green fluorescent proteins; in situ hybridization; lethal genes; major histocompatibility complex; northern blottings; phenotype; yeast two hybrid system; zebrafish

DESCRIPTION (provided by applicant): This proposal is to complete an ongoing mapping of genes involved in axis defects in mice and move on to study a new and cloned zebrafish neurogenic mutation. One ultimate goal is to make a model of it in mouse. To date, a number of model organisms have been used in the study of molecular development: those that have really good genetics or ones for which developing stages are readily accessible but for technical reasons genetic studies are not an option. Recently, zebrafish have come into use because it satisfies both criteria. Among its advantages are that it is a vertebrate, hundreds of embryos can be obtained in a day, the embryos are transparent and can be easily visualized. Thus, novel phenotypes in early neural development are evident whereas they would be inaccessible in a mammal. In a retroviral mutant screen in zebrafish, a mutation has a viral tag that can be used to easily clone it. From the Hopkins screen at MIT, I have chosen to analyze a recessive lethal (Hi904) that causes disorganization and tumor-like overgrowth of the nervous system. The gene has been cloned and is novel. We will do experiments that are easily accomplished in fish, and the rest in mice where we can recreate the mutation by homologous recombination. The experiments in zebrafish will include: expression studies of Hi904 message in embryos, analysis with well-characterized molecular markers to assess cell number in the nervous system of mutant embryos, and also patterning and state of differentiation. This will be accomplished by in situ hybridization and antibody staining. The experiments in mice will include: obtaining the complete mouse cDNAs including expected alternative splice forms, expression analysis in normal and tumor tissues and in situ hybridization in embryos. We will address tumorigenicity by transplanting mutant fish cells into nude mice. We will make antibody for functional studies. Mutations in mouse will be generated using homologous recombination. The biochemical and cellular function of Hi904 protein will be explored by studying its localization and interactions. It is expected that this set of genes will define a new tumor suppressor that plays-a role in the development of primitive neuroectodermal tumors of children.

描述（由申请人提供）：该提案是为了完成持续的 涉及小鼠轴缺陷的基因的映射并继续研究新的 并克隆斑马鱼神经源突变。一个最终目标是制作模型 它在鼠标中。迄今为止，已经使用了许多模型生物 分子发育研究：那些真正具有良好遗传学或遗传学的人 对于哪个开发阶段很容易访问，但出于技术原因 遗传研究不是一种选择。最近，斑马鱼已经使用了 因为它满足这两个标准。它的优势包括它是 脊椎动物，数百个胚胎可以在一天内获得，胚胎是 透明，可以轻松可视化。因此，早期的新表型 神经发育是显而易见的，而在哺乳动物中它们将无法获得。 在斑马鱼的逆转录病毒突变屏中，突变具有一个病毒标签，可以 被用来轻松克隆它。在麻省理工学院的霍普金斯屏幕上，我选择了 分析导致混乱和类似肿瘤的隐性致命（HI904） 神经系统的过度生长。该基因已被克隆，是新颖的。我们 将进行轻松在鱼中完成的实验，其余的小鼠 我们可以通过同源重组来重现突变。实验 在斑马鱼中，将包括：HI904消息的表达研究， 用良好的分子标记物分析，以评估细胞数 突变胚胎的神经系统，也有图案和状态 分化。这将通过原位杂交和 抗体染色。小鼠的实验将包括：获得完整 小鼠cDNA包括预期的替代剪接形式，表达分析 正常和肿瘤组织以及胚胎中的原位杂交。我们将解决 通过将突变鱼细胞移植到裸鼠中，致肿瘤。我们会做的 功能研究的抗体。鼠标中的突变将使用 同源重组。 HI904的生化和细胞功能 蛋白质将通过研究其本地化和相互作用来探索蛋白质。这是 预计这组基因将定义一个播放的新肿瘤抑制剂-a 在儿童原始神经皮质肿瘤的发展中的作用。