CYSTIC DILATATION OF NEPHRONS IN TRANSGENIC INV MICE

转基因 INV 小鼠肾单位囊性扩张

• 批准号: 6262576
• 负责人: CARRIE L PHILLIPS
• 金额: $ 12.07万
• 依托单位: INDIANA UNIV-PURDUE UNIV AT INDIANAPOLIS
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-07-01 至 2006-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6262576
• 关键词: autosomal recessive trait; chimeric proteins; developmental genetics; disease /disorder model; electron microscopy; embryogenesis; genetically modified animals; green fluorescent proteins; immunofluorescence technique; laboratory mouse; light microscopy; mammalian embryology; messenger RNA; polycystic kidney; postpartum; renal tubule

Polycystic kidney disease (PKD) is the most common inherited renal disease, accounting for 5-10% of end-stag renal disease with an annual cost of $240,000,000. The etiology of renal cysts is especially important for the children an adults with inherited PKD who may require dialysis or transplantation for survival. We have a newly describe transgenic mouse model of PKD, called inv/inv, which will be used to conduct studies not possible in humans. The long term goal of this project is to understand how the inheritance of a novel situs inversus gene, inv, results in PKD and loss of renal function. Our general strategy is to study the kidneys of inv/inv mice, which share phenotypic features with autosomal recessive (AR) PKD. They develop situs inversus, PKD and die prematurely from renal failure. The inv gene has been fully cloned and sequenced. Our preliminary data suggest inv is an intracellular protein with discrete localization in specific segments of the nephron. Our experiments will test the hypothesis that the inv gene codes for a developmentally important protein that maintains nephron integrity. Grant funding is a major factor in the PI's immediate goal of attaining research independence and the long-term career goal of becoming a successful and productive investigator in an academic institution. The specific aims of this present study are as follows: 1. To characterize the inv/inv mouse model of ARPKD, we will perform single and dual photon fluorescence microscopy with cell-specific antibodies and lectins. Immunogold electron microscopy with anti-inv antibodies will target the intracellular localization of the inv protein. These microscopy experiments will allow us to appropriately direct mechanistic studies of inv protein function and interactions. 2. In situ hybridization studies will be used to determine the cellular levels of inv messenger RNA during critical periods of development. This is especially important, as incremental variations in mRNA expression are known to occur during narrow developmental windows. 3. We will use green fluorescent protein-inv constructs and anti-inv antibodies t establish spatial and temporal inv expression patterns during development. We believe the inv/inv model offers a new an exciting opportunity to extend the understanding of the genetic mechanisms of nephrogenesis and the abnormalities in this process which lead to PKD.

多囊性肾脏疾病（PKD）是最常见的肾脏疾病，占末端纳尔肾病的5-10％，年费用为240,000,000美元。肾囊肿的病因对于可能需要透析或移植才能生存的儿童尤其重要。我们有一个新的PKD的转基因小鼠模型，称为Inv/Inv，该模型将用于进行人类进行研究。该项目的长期目标是了解新型Situs Inversus基因的遗传如何导致PKD和肾功能的丧失。我们的一般策略是研究Inv/Inv小鼠的肾脏，这些肾脏与常染色体隐性（AR）PKD共享表型特征。他们会因肾衰竭而过早地发展现场倒置，PKD并过早地死亡。 INV基因已被完全克隆和测序。我们的初步数据表明INV是一种细胞内蛋白，在肾单位的特定段中具有离散定位。我们的实验将检验以下假设：INV基因代码为维持肾单位完整性的发育重要蛋白质代码。赠款资金是PI取得研究独立性的近期目标的主要因素，也是成为学术机构中成功和生产性研究人员的长期职业目标。本研究的具体目的如下：1。为了表征ARPKD的Inv/Inv小鼠模型，我们将用细胞特异性抗体和凝集素执行单个和双光子荧光显微镜。具有抗INV抗体的免疫元电子显微镜将针对Inv蛋白的细胞内定位。这些显微镜实验将使我们能够适当地直接对INV蛋白功能和相互作用的机械研究。 2。原位杂交研究将用于确定在关键发展期间INV Messenger RNA的细胞水平。这尤其重要，因为已知在狭窄的发育窗口中已知MRNA表达的增量变化。 3。我们将使用绿色荧光蛋白INV构建体和抗INV抗体t在发育过程中建立空间和时间INV表达模式。我们认为，INV/INV模型为一个新的激动人心的机会扩展了对肾脏发生的遗传机制和此过程中导致PKD的异常的理解。