RENAL CYSTIC DISEASE--CHARACTERIZING THE MOUSE CPK GENE

导致肾囊肿的突变提供了强有力的工具， 肾囊肿的病因有哪些？在 人类遗传性囊性肾病是遗传的， 孟德尔性状，表现出广泛的变异表型和帐户 对于成人和成人中约10%的终末期肾病， 儿科人群。 连锁研究表明， 不同的位点引起肾囊肿形成。 然而，只有一些人类 基因座已被定位，但迄今为止还没有克隆。 老鼠提供了一个很好的实验模型来识别 肾囊肿的治疗方法有哪些 在小鼠中， 已经描述了囊性肾突变，每种突变都破坏了一种独特的 基因和突变表型与人类疾病非常相似。 的 在这些模型中，先天性多囊肾（cpk）突变是最好的 表征了 基于显微切割和细胞生物学数据，我们 假设CPK是两者重要分子决定因子 肾小管的终末分化 上皮细胞 作为定位克隆策略的第一步，我的实验室 将cpk定位于近端小鼠染色体12。 值得注意的是，没有重组 在cpk和D12 Nyu 2之间检测到事件。 我们的基因图谱： 着丝粒-（Odc，D12 Mit 10）-（cpk，D12 Nyu 2）-（Tpo，D12 Mit 12）-端粒， 位置cpk在以D12 Nyu 2为中心的1.3cM区域内。 基于 这张图，我们将建立克隆所需的分子框架， cpk基因 本提案的目标是：1）建立一个 以cpk为中心的染色体区域的物理图， 脉冲场凝胶电泳技术与酵母 人工染色体（YAC）克隆; 2）鉴定候选cpk cDNA 使用HTF岛识别的互补策略，YAC- 基于cDNA选择和mRNA差异显示和3）表征 候选cpk cDNA。 一旦cpk基因被克隆，未来的研究将针对1） 功能上表征了这种定义明确的 PKD的小鼠模型;和2）确定肾间质与PKD的关系。 肾小管囊肿形成和肾小管分化。 鉴于 小鼠和人类之间广泛的遗传保守性，我们将 还使用cpk基因来鉴定其人类同源物， 这一人类基因在肾囊肿疾病中的作用。

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.