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Molecular pathegenesis of congenital renal disease

先天性肾病的分子发病机制

基本信息

批准号：
14571016
负责人：
KAWAHARA Michio
金额：
$ 2.24万
依托单位：
Tokyo Medical and Dental University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2002
资助国家：
日本
起止时间：
2002 至 2003
项目状态：
已结题

项目摘要

1. We previously reported an AQP2 gene mutation (763 -772de1) causing autosomal dominant nephrogenic diabetes insipidus (NDI). To determine the cellular pathogenesis of the NDI, MDCK cells were transfected with the wild-type AQP2, or the 763-772de1, or both. Immunofluorescence microscopy showed that the wild-type AQP2 and the 763-772de1 were expressed in the apical and basolateral region, respectively. When they were cotransfected, mixed oligomers of the wild-type AQP2 and the 763-772de1 were mistargeted to the basolateral membrane due to the dominant negative effect of the mutant. This defect is likely to explain the pathogenesis of autosomal dominant NDI.2. To determine we the role of AQP2 C-terminus for apical trafficking, C-terminal mutants of AQP2 were transfected into MDCK cells to determine the part responsible for apical trafficking. Our Tresults suggest that the last 10 amino acids are the determinant for apical trafficking of AQP2.3. We identified two novel AQP2 gene mutation (Q57P, GlOOV) causing autosomal recessive NDI. When these mutant AQP2s were expressed in Xenopus oocytes, they were retained in intracellular compartment, suggesting the NM is caused by the trafficking defect.4. We examined the osmotic regulation of AQP1 expressed in cultured mesothelial cells in peritoneum. Exposure to hyperosmolality significantly increased the expression of AQP1 and cell water penneability after 24 hours, suggesting upregulation of AQP1 by hyperosmolality.5. We cloned a novel water channel, AQP1-. AQP10 was specifically expressed in ileum. Xenopus oocyte expression study revealed that AQP1O permeates glycerol in addition to water.

1.我们以前曾报道过一个AQP 2基因突变（763 - 772 de 1）引起的常染色体显性遗传性肾性尿崩症（NDI）。为了确定NDI的细胞发病机制，用野生型AQP 2或763- 772 de 1或两者转染MDCK细胞。免疫荧光显微镜显示野生型AQP 2和763- 772 de 1分别在顶侧和基底侧区域表达。当它们被共转染时，由于突变体的显性负效应，野生型AQP 2和763- 772 de 1的混合寡聚体被错误定位到基底外侧膜。这种缺陷可能解释常染色体显性遗传NDI的发病机制。为了确定AQP 2 C-末端在顶端运输中的作用，将AQP 2的C-末端突变体转染到MDCK细胞中以确定负责顶端运输的部分。我们的结果表明，最后10个氨基酸是决定AQP2.3的顶端运输。我们发现了两个新的AQP 2基因突变（Q57 P，GlOOV）导致常染色体隐性NDI。当这些突变的AQP 2在非洲爪蟾卵母细胞中表达时，它们被保留在细胞内隔室中，表明NM是由运输缺陷引起的。4.我们研究了水通道蛋白1在腹膜间皮细胞中表达的渗透调节作用。暴露于高渗24小时后，AQP 1的表达和细胞水渗透性显著增加，提示高渗可上调AQP 1的表达.我们克隆了一种新的水通道，AQP 1-。AQP 10在回肠特异性表达。非洲爪蟾卵母细胞表达研究表明，AQP 1 O渗透甘油除了水。