DLGH1 AND UROGENITAL DEVELOPMENT

DLGH1 和泌尿生殖系统发育

基本信息

批准号：
7876620
负责人：
JEFFREY H MINER
金额：
$ 31.98万
依托单位：
WASHINGTON UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2008
资助国家：
美国
起止时间：
2008-08-14 至 2013-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7876620
关键词：
Affect Alleles Animals Binding Proteins Biological Process Biology Cell Line Cell Polarity Cell Proliferation Cells Complex Cytoskeleton Data Defect Development Drosophila Proteins Drosophila genus Embryo Employee Strikes Epithelial Epithelial Cells Epithelium Event Exhibits Eyelid structure Family Family member Female Fibroblasts Gene Expression Genes Genetic Genitourinary system Golgi Apparatus Guanylate kinase Hand Health Homologous Gene Human Hydronephrosis Immigration Immunofluorescence Immunologic In Vitro Kidney Kidney Diseases Knock-out Knockout Mice Life MAPK14 gene Mammalian Cell Mediating Mesenchymal Mesenchyme Metanephric Diverticulum Methods Molecular Molecular Profiling Morphogenesis Morphology Motion Mus Muscle Mutagenesis Mutant Strains Mice Mutate Mutation Nature Palate Pattern Peristalsis Phenotype Phosphotransferases Play Population Prevention Protein Binding Domain Protein Family Proteins Regulation Reporter Research Role SH3 Domains Scaffolding Protein Seminal Vesicles Signal Transduction Site Smooth Muscle Smooth Muscle Myocytes Staging Sternum Stromal Cells Structure Structure of mesonephric duct Suppressor-Effector T-Lymphocytes System Testing Time Tissues Transfection Transgenes Transgenic Mice Tumor Suppressor Proteins Ureter Urinary tract Urine Urothelial Cell Urothelium Vagina Work Wound Healing base cell motility cell type dlg protein fly glycogen synthase kinase 3 beta human DLG1 protein imaginal disc in vivo male member membrane-associated guanylate kinase migration mouse model mutant novel null mutation postsynaptic density protein public health relevance reproductive research study synaptic function urinary tract obstruction

项目摘要

DESCRIPTION (provided by applicant): The long term objective of this research involves determining the mechanism whereby the mouse discs- large homolog 1 gene, Dlgh1, regulates urogenital development. DLGH1 is a scaffolding protein with multiple protein-protein interaction domains, including three PDZ domains and one SH3 domain. Dlgh1 is a homolog of the Drosophila discs-large gene, dlg, which is a founding member of the eponymous PDZ family of proteins and also of the MAGUK (membrane-associated guanylate kinase) family. Studies in Drosophila suggest that dlg is a tumor suppressor, because mutation of dlg causes overgrowth of imaginal discs. Studies in both fly and mammalian cells suggest that dlg and its homologs are also involved in establishing and/or maintaining epithelial cell polarity. We have studied mice lacking DLGH1 and found several urogenital defects, including sporadic renal agenesis, congenital hydronephrosis, and reproductive tract abnormalities. We have shown that hydronephrosis is associated with dramatic smooth muscle alignment defects; the normally circular muscle aberrantly aligns in the longitudinal direction. This greatly impairs the squeezing motion of peristalsis, such that urine is retained in the kidney. In addition, ureteric stromal cells, which normally lie between the urothelium and the smooth muscle layers and express Raldh2, are absent from Dlgh1 mutant ureters. The absence of these cells could also affect ureteric function, either directly or indirectly. Immunofluorescence studies show that DLGH1 is expressed strongly in the urothelium and weakly in other cells of the ureter. We will determine which cellular compartment must express Dlgh1 for normal smooth muscle cell alignment and for differentiation and/or migration of ureteric stromal cells using a new floxed Dlgh1 allele and appropriate Cre transgenic mice. In addition, we will generate chimeric mice to investigate whether DLGH1 acts in a non-cell autonomous fashion. Morphological and functional assessments will be performed to determine the cellular requirements for Dlgh1 expression. Next we will use genetic methods to investigate the hypothesis that the interaction of other DLGH family members with known or suspected DLGH1 binding proteins are compensating in part for the absence of DLGH1. Finally, we will use in vitro methods to define the role of DLGH1 in cell migration, polarization, and signaling. PUBLIC HEALTH RELEVANCE: Congenital urinary tract abnormalities are a relatively common health problem in the human population and are responsible for a significant number of cases of progressive renal disease. We have generated a novel mouse model of hydronephrosis (Dlgh1-/- mice) involving the misalignment of ureteric smooth muscle, which is commonly found in human cases of urinary tract obstruction. A better understanding of the biology of DLGH1 could have important implications for understanding and perhaps for treatment or prevention of urinary tract abnormalities in humans.

描述（由申请人提供）：这项研究的长期目标涉及确定小鼠椎间盘 - 大型同源物1基因DLGH1的机制，可调节泌尿生殖器的发育。 DLGH1是一种具有多种蛋白质 - 蛋白质相互作用结构域的脚手架蛋白，包括三个PDZ结构域和一个SH3结构域。 DLGH1是果蝇圆盘大基因DLG的同源物，该基因是同名PDZ蛋白质家族的创始成员，也是Maguk（膜相关的鸟苷酸盐激酶）家族的植物。果蝇的研究表明，DLG是肿瘤抑制因子，因为DLG的突变会导致想象盘的过度生长。在FLY和哺乳动物细胞中的研究表明，DLG及其同源物也参与建立和/或维持上皮细胞极性。我们研究了缺乏DLGH1的小鼠，并发现了几种泌尿生殖器缺陷，包括零星的肾脏发育不全，先天性肾结通和生殖道异常。我们已经表明，肾积水与剧烈的平滑肌比对缺陷有关。通常，圆形肌肉异常对准纵向方向。这极大地损害了蠕动的挤压运动，因此肾脏保留在肾脏中。此外，DLGH1突变输尿管中不存在通常位于尿路上皮和平滑肌层和表达RALDH2之间的输尿管基质细胞。这些细胞的不存在也可能直接或间接影响输尿管功能。免疫荧光研究表明，DLGH1在尿路上皮中强烈表达，在输尿管的其他细胞中弱表示。我们将使用新的Floxed DLGH1等位基因和适当的CRE转基因小鼠来确定哪种细胞室必须表达正常平滑肌细胞对齐的DLGH1和输尿管基质细胞的分化和/或迁移。此外，我们还将生成嵌合小鼠，以研究DLGH1是否以非细胞自主方式起作用。将进行形态学和功能评估，以确定DLGH1表达的细胞需求。接下来，我们将使用遗传学方法来研究以下假设：其他DLGH家族成员与已知或怀疑的DLGH1结合蛋白的相互作用部分是由于缺乏DLGH1的。最后，我们将使用体外方法来定义DLGH1在细胞迁移，极化和信号传导中的作用。公共卫生相关性：先天性尿路异常是人口中相对常见的健康问题，并负责大量进行性肾脏疾病病例。我们已经产生了一种新型的肾结化小鼠模型（DLGH1 - / - 小鼠），涉及输尿管平滑肌的未对准，这在人类尿路阻塞的情况下通常发现。对DLGH1生物学的更好理解可能对人类的理解或预防尿路异常具有重要意义。