MOLECULAR GENETIC STUDIES OF POLYCYSTIC KIDNEY DISEASE

有几条证据表明基底膜的改变 结构可能是观察到的临床和形态变化的基础 成人多囊肾病(APKD)。建议的研究是设计的 要回答这个问题：在一个或多个编码基因中发生突变 基底膜蛋白[IV型胶原层粘连蛋白、巢蛋白、牙本质蛋白或 硫酸乙酰肝素蛋白多糖(HSPG)核心蛋白]病因(APKD)？这个 提出了以下方法：(1)组织结构 APKD中编码主要基底膜蛋白的基因 种群数量将通过Southern杂交分析来确定。不寻常的图案 将与在控件面板中看到的控件进行比较，以确定 前者是严重的基因改变的结果。人PRO基因的c DNA探针 α1(IV)胶原蛋白和层粘连蛋白链B1目前可用于这些 学习。小鼠层粘连蛋白A和B2、Nidogen、entactin和HSPG基因 核心蛋白将用于分离同源的人cDNAs探针。(2) 这些基因结构中更细微的变化将通过 连锁分析。有多个受影响个人的大家庭将 对APKD表型和特异性基因进行共分离检查 等位基因的遗传是通过多态限制酶来检测的。 三个大的、多代的家庭适合进行这些研究； 其他家庭将在#年的早期阶段确定并确定特征 提议的项目。(3)第(1)款或第(1)款所指的个人或家庭 (2)表明感兴趣的基因之一发生突变，对mrna的研究。 由该基因编码的基因将被执行以精确定位涉及 突变。(4)由(1)或(2)确定的突变基因将是 克隆并测序，以证明突变的性质。(5) 将合成针对突变和正常等位基因的寡核苷酸探针 并用于研究多个家庭，以努力确定 APKD的遗传异质性。

Several lines of evidence suggest that alterations of basement membrane structure may underlie the clinical and morphologic changes observed in adult polycystic kidney disease (APKD). The proposed studies are designed to answer the question: Do mutation in one or more of the genes encoding basement membrane proteins [Type IV collagen laminin, nidogen, entactin, or heparan sulfate proteoglycan (HSPG) core protein] cause (APKD)? The following approaches are proposed: (1) The structure and organization of the genes encoding the major basement membrane proteins in an APKD population will be determined by Southern blot analysis. Unusual patterns will be compared to those seen in a panel of controls to determine if the former are the result of gross gene alterations. Human cDNA probes for pro Alpha1(IV) collagen and laminin chain B1 are currently available for these studies. Murine genes for laminin A and B2, nidogen, entactin, and HSPG core protein will be used to isolate the homologous human cDNA probes. (2) More subtle alterations in the structure of these genes will be detected by linkage analysis. Large families with multiple affected individuals will be examined for cosegregation of a the APKD phenotype and specific gene alleles whose inheritance is detected by polymorphic restriction sites. Three large, multi-generation families are suitable for these studies; other families will be ascertained and characterized in the early phase of the proposed project. (3) In individual or families in whom either (1) or (2) indicates mutation in one of the genes interest, studies of mRNA encoded by this gene will be performed to pinpoint the region involved in the mutation. (4) Mutant genes identified by either (1) or (2) will be cloned and sequenced to demonstrate the nature of the mutation. (5) Oligonucleotide probes will be synthesized for mutant and normal alleles and used to study multiple families in an effort to determine the extent of genetic heterogeneity in APKD.