DYSGENIC KIDNEY LETHAL MUTANT MICE

我们研究的目标是确定导致这种缺失的基因 在我们最近培育的一种嵌入型小鼠突变中的肾脏 并了解它们在肾脏发育中的作用。突变的基因座有 被称为KDL的肾脏：发育不良，新生儿死亡(KDL)。这 突变被鉴定为由转基因引起的插入突变。 整合。纯合子小鼠出生后24小时内死亡， 肾脏发育异常。大体组织学检查未发现 其他组织的异常。我们有分离出的序列位于 并鉴定了一个编码新基因的基因 G蛋白偶联受体，该受体已被插入打乱。 我们已经从大鼠中克隆了受体的cdna，并对其进行了测序。 完全是。该基因(在此称为KDLR)被表达 主要在成年小鼠的肾脏中发现，并在整个小鼠中发现 胚胎RNA。kdlr基因的氨基末端是 并删除和KDL小鼠。跨越已删除区域的PAC克隆具有 是从野生型小鼠KDLR区域分离出来的，将对其进行评估 寻找其他候选基因。初步测绘的插入物表明 最大缺失区域小于100kb，因此 可能只含有KDLR，或者可能含有数量有限的其他基因。 我们研究的具体目标包括：1)完成地图绘制 Kd1插入位点和mKDLR野生型基因组座位：扫描， 其他候选基因的鉴定和评估；2)提供 KDL突变小鼠和KDLR缺失小鼠缺陷的详细分析 包括免疫组织化学、超微结构和形态计量 刻画。3)制作靶向消融KDLR的小鼠，并 并与KDL表型进行比较；KDLRr是一种 优秀的候选人，因此它将与 绘制该地区的地图。4)组织特异性和发育性分析 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)调查职能部门 KDLR OR的配体结合和细胞活性的表征 其他候选基因。这些研究将导致对 新基因的功能如何影响肾脏发育并提供 对肾脏形成过程的洞察。

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.