DYSGENIC KIDNEY LETHAL MUTANT MICE

我们研究的目标是确定导致缺乏的基因 我们最近制造的一个插入突变小鼠的肾脏 并了解它们在肾脏发育中的作用。突变基因座具有 被称为kdl，用于肾脏：发育不良的新生儿致死性（kdl）。这 突变被鉴定为由于转基因引起的插入突变 一体化纯合子小鼠在出生后24小时内死亡， 肾脏发育异常大体组织学检查显示， 其他组织的异常。我们有孤立的序列 插入位点的转基因，并确定了一个基因编码的新的 G蛋白偶联受体，其已被插入破坏。 我们已经克隆了大鼠受体的cDNA并进行了测序 完全是该基因（在此称为KDLR）表达于 主要存在于成年小鼠肾脏中， 胚胎RNA早在第7天。kdlr基因的氨基末端是 缺失和KDL小鼠。跨删除区域的PAC克隆具有 从野生型小鼠KDLR区域分离，并将进行评价 其他候选基因。插入物的初步映射表明 最大缺失区域小于100 kb，因此 可能仅含有KDLR或可能含有有限数量的其他基因。 本研究的具体目标包括：1）完成对 kdl插入位点和mKDLR野生型基因组基因座：扫描， 其他候选基因的鉴定和评估; 2）提供一个 详细分析kdl突变小鼠和KDLR缺失小鼠中的缺陷 包括免疫组织化学、超微结构和形态计量学 特征化。3)产生具有KDLR靶向消融的小鼠， 其他候选基因，并与kdl表型进行比较; KDLRr是一个 优秀的候选人，因此，它将同时进行测试， 该地区的地图。4)组织特异性和发育分析 包括KDLR在内的候选基因的表达;并确定人类 KDLR的染色体定位。5)调查功能 KDLR或KDLR的配体结合和细胞活性的表征 其他候选基因这些研究将有助于了解 新基因的功能如何影响肾脏发育， 深入了解肾发生的过程

The goal of our research is to define the genes responsible for the lack of kidneys in an insertional mouse mutant that we have recently produced and to understand their role in kidney development. The mutant locus has been termed kdl for kidney: dysgenic, neonatal lethal (kdl). This mutation was identified as an insertional mutation due to a transgene integration. Homozygous mice die within 24 hours of birth, with abnormally developed kidneys. Gross histological examination reveals no abnormalities in other tissues. We have isolated sequences flanking the insertion site of the transgene and identified a gene encoding a novel G-protein coupled receptor, which has been disrupted by the insertion. We have cloned the cDNA of the receptor from rat and sequenced it entirely. This gene (hereby referred to as KDLR) is expressed predominantly in the adult mouse kidney and is identified in whole mouse embryo RNA as early as day 7. The amino terminus of the kdlr gene is deleted and kdl mice. A PAC clone which spans the deleted region has been isolated from the wild type mouse KDLR region and will be evaluated for other candidate genes. Preliminary mapping of the insert indicates that the maximum deleted region is less than 100 kb, and consequently likely to contain only KDLR or possibly a limited number of other genes. The specific aims of our study include: 1) Complete the mapping of the kdl insertion site and the mKDLR wild type genomic loci: scanning, identification, and evaluation of other candidate genes; 2) Provide a detailed analysis of defects in kdl mutant mice and KDLR null mice including immunohistochemical, ultrastructural and morphometric characterizations. 3) Produce mice with targeted ablation of KDLR and other candidate genes and compare with the kdl phenotype; KDLRr is an excellent candidate, hence it will be tested concurrently with the mapping of the region. 4) Analysis of tissue specific and developmental expression of candidate genes including KDLR; and determine the human chromosomal location for KDLR. 5) Investigate the functional characterization of ligand binding and cellular activity of KDLR or other candidate genes. These studies will lead to an understanding of how the functions of new genes affect kidney development and provide an insight into the process of nephrogenesis.