RENAL CYSTIC DISEASE--CHARACTERIZING THE MOUSE CPK GENE

导致肾囊性疾病的突变提供了强大的工具 确定肾囊肿形成的分子决定因素。在……里面 人类，遗传性囊性肾病是遗传性的 孟德尔性状，在表型和账户上表现出很大的差异 约10%的终末期肾脏疾病在成人和 儿科人口。连锁研究表明，几个基因的突变 不同的基因座可导致肾囊性变。然而，只有部分人类 已经绘制了基因座图谱，到目前为止还没有克隆到任何一个。 小鼠提供了一个很好的实验模型来识别 肾囊肿形成的分子决定因素。在鼠标中，有几个 囊性肾突变已被描述为每一种破坏不同的 基因和突变的表型与人类疾病非常相似。的 这些模型中，先天性多囊肾(CPK)突变最好 特色化的。基于显微解剖和细胞生物学数据，我们 假设CPK是两者的重要分子决定因素 肾小管的成囊与终末分化 上皮细胞。 作为定位克隆策略的第一步，我的实验室已经 将CPK定位于小鼠12号染色体近端。值得注意的是，没有重组 在CPK和D12Nyu2之间检测到事件。我们的基因图谱： 着丝粒-(ODC，D12Mit10)-(CPK，D12Nyu2)-(TPO，D12-12)-端粒， 将CPK放置在以D12Nyu2为中心的1.3厘米区域内。基于 在这张图上，我们将建立克隆所需的分子框架 CPK基因。这项提议的目标是：1)建立一个 以CPK为中心的染色体区域的物理图谱 脉冲场凝胶电泳(PFGE)和酵母菌技术 人工染色体(YAC)克隆；2)鉴定CPK候选cDNA 利用HTF岛屿识别的补充战略，YAC- 基于c DNA选择和m RNA差异显示及3)特征分析 候选人CPK的cDNA。 一旦CPK基因被克隆，未来的研究将针对1) 在功能上表征了这个定义明确的分子缺陷 建立小鼠PKD模型；2)确定肾损害与肾功能损害的关系。 肾小管囊肿形成与肾小管分化。根据…… 老鼠和人类之间广泛的遗传保守性，我们将 也使用CPK基因来鉴定其人类同源基因并确定 这个人类基因在肾囊性疾病中的作用。

Mutations that cause renal cystic disease provide powerful tools to identify the molecular determinants of renal cyst formation. In humans, genetic cystic kidney diseases are inherited as simple mendelian traits, exhibit wide variability in phenotype and account for approximately 10% of end stage renal disease in both adult and pediatric populations. Linkage studies that mutations in several distinct loci cause renal cystogenesis. yet, only some of the human loci have been mapped and to date none have cloned. The mouse provides an excellent experimental model to identify molecular determinants of renal cyst formation. In the mouse, several cystic kidney mutations have been described each disrupts a distinct gene and the mutant phenotypes closely resemble human diseases. Of these models, the congenital polycystic kidney (cpk) mutation is best characterized. Based on microdissection and cell biology data, we hypothesize that cpk is an important molecular determinant of both cystogenesis and the terminal differentiation of renal tubular epithelia. As the first step in a positional cloning strategy, my laboratory has mapped cpk to proximal mouse Chromosome 12. Of note, no recombination events were detected between cpk and D12Nyu2. Our genetic map: centromere-(Odc,D12Mit10)-(cpk,D12Nyu2)-(Tpo,D12Mit 12)-telomere, positions cpk within a 1.3 cM region centered on D12Nyu2. Based on this map, we will establish the molecular framework required to clone the cpk gene. The goals of this proposal are to: 1) construct a physical map of the chromosomal region centered on cpk using the techniques of pulsed field gel electrophoresis (PFGE) and yeast artificial chromosome (YAC) cloning; 2) identify candidate cpk cDNAs using the complementary strategies of HTF island identification, YAC- based cDNA selection and mRNA differential display and 3) characterize the candidate cpk cDNAs. Once the cpk gene is cloned, future studies will be directed at 1) functionally characterizing the molecular defect in this well-defined mouse model of PKD; and 2) determining the relationship between renal tubular cyst formation and renal tubular differentiation. In light of the extensive genetic conservation between mice and humans, we will also use the cpk gene to identify its human homolog and determine the role of this human gene in renal cystic disease.