Kidney development and cystogenesis in Medaka

青鳉的肾脏发育和囊肿发生

• 批准号: 6856354
• 负责人: TOMOKO OBARA
• 金额: $ 15.15万
• 依托单位: CASE WESTERN RESERVE UNIVERSITY
• 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 2006-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6856354
• 关键词: autosomal dominant trait; disease /disorder model; gene expression; gene mutation; genetically modified animals; medaka; messenger RNA; nephrogenesis; polycystic kidney; polymerase chain reaction; protein protein interaction; renal tubule

DESCRIPTION (provided by applicant): Autosomal dominant polycystic kidney disease (ADPKD) is a hereditary disease occurring at a frequency of 1:1000 in humans and characterized by cyst formation in kidney tubules, deregulated fluid transport and alteration of extracellular adhesion. ADPKD is caused by mutations in the PKD1 or PKD2 gene that encode polycystin-1 and polycystin-2, respectively. The primary causes and mechanisms of cyst formation in ADPKD remain elusive. Our attempts to knockdown pkdl in zebrafish failed because of gene redundancy. Medaka is a unique inbred aquatic vertebrate fish model system, which shows lower redundancy in gene number compared to zebrafish. The use of medaka provides an opportunity to develop a novel system to study organogenesis and to understand the mechanisms of cystogenesis. Given that PKD1 mutations account for 85% of all ADPKD cases in humans, we have targeted pkdl in medaka to understand its function(s). Preliminary results demonstrate that targeting pkdl in medaka (in contrast to zebrafish) causes prronephric cysts formation. We have also established a system using medaka for studies of kidney, cystogenesis that is sensitive for the defect of polycystin-1 interacting proteins such as polycystin-2, which can be used to assess the in vivo relevance of the large number of polycystin-1 interacting proteins. In specific aim 1, we will test the in vivo function of specific polycystin-1 motifs by disrupting pkdl mRNA splicing and generate an allelic series of deletions in medaka. For this purpose, we propose to use our urogenital specific medaka transgenic GFP line to monitor the progress of cyst formation, and to analyze the kidney phenotypes as well as other potential defects in organogenesis caused by the defers. In specific aim 2, we will validate the biological significance of polycystin-1 interacting proteins.

描述（由申请方提供）：常染色体显性多囊肾病（ADPKD）是一种遗传性疾病，在人类中的发生率为1：1000，其特征为肾小管囊肿形成、液体转运失调和细胞外粘附改变。ADPKD是由分别编码多囊蛋白-1和多囊蛋白-2的PKD 1或PKD 2基因突变引起的。ADPKD囊肿形成的主要原因和机制仍不清楚。由于基因冗余，我们在斑马鱼中敲除pkdl的尝试失败了。青鳉是一种独特的近交水生脊椎动物模式鱼，与斑马鱼相比，其基因冗余度较低。青鳉的使用提供了一个机会，开发一个新的系统来研究器官发生和了解的机制，cystogenesis。鉴于PKD 1突变占人类所有ADPKD病例的85%，我们已经靶向青鳉中的pkdl以了解其功能。初步结果表明，针对青鳉（与斑马鱼相反）的pkdl导致原肾囊肿的形成。我们还建立了一个系统，使用青鳉的研究肾脏，cystogenesis是敏感的缺陷的多囊蛋白-1相互作用的蛋白质，如多囊蛋白-2，这可以用来评估在体内的相关性的大量的多囊蛋白-1相互作用的蛋白质。 在具体目标1中，我们将通过破坏pkdl mRNA剪接并在青鳉中产生一系列等位基因缺失来测试特定多囊蛋白-1基序的体内功能。为此，我们建议使用我们的泌尿生殖系统特异性青鳉转基因GFP系来监测囊肿形成的进展，并分析肾脏表型以及由延迟引起的器官发生中的其他潜在缺陷。在具体目标2中，我们将验证多囊蛋白-1相互作用蛋白的生物学意义。