Electrophysiological Analysis of Drosophila PKD2

果蝇 PKD2 的电生理分析

• 批准号: 6440152
• 负责人: XIANGYI LU
• 金额: $ 14.35万
• 依托单位: UNIVERSITY OF ALABAMA AT BIRMINGHAM
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-09-30 至 2003-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6440152
• 关键词: Drosophilidae; biotechnology; calcium ion; cations; confocal scanning microscopy; electrophysiology; epithelium; gene expression; genetic models; genetically modified animals; membrane channels; polycystic kidney; regulatory gene; technology /technique development; voltage /patch clamp; voltage gated channel

DESCRIPTION (provided by applicant): Much progress has been made recently in understanding the autosomal dominant polycystic kidney disease (ADPKD), which affects millions of people world-wild. The main disease phenotype is characterized by the persistent proliferation and de-differentiation of renal epithelial cells that eventually form fluid-filled cysts. Gradually, these cysts cause anatomical distortion and kidney failure. One of the disease-causing genes, PKD2, has a significant overall sequence and structure conservation to various known cation channels. Recent study indicates that expression of PKD2 or a related protein PKDL is sufficient to produce Ca2+-permeable non-selective cation channel activities. Another ADPKD disease gene, PKD1, is also a transmembrane protein that has been shown to interact with PKD2. PKD 1 may regulate PKD2 channel activity which initiates yet undefined intracellular signal transduction pathway that maintains proper epithelial cell polarity, secretion and other functions. This proposal applies newly developed tools of functional genomics to the study of PKD2 gene through its homologue in Drosophila, namely the DmPKD2 gene. We have cloned the DmPKD2 gene and it shows similarities of 48 percent and 59 percent to human PKD2 in two most conserved regions that contain the predicted transmembrane segments. The N- and C-termini of DmPKD2.are more divergent. We propose here to test the hypothesis that DmPKD2 functions as a channel as was shown for other vertebrate PKD2-related proteins. The properties and regulation of DmPKD2 will be quantified. We also propose to test the channel function of DmPKD2 in its native environment, namely living fly epithelial tissues. Specifically, we will genetically over-express or ablate the production of DmPKD2 in culture cells (in vitro) and fly tissues (in vivo). Single channel analyses will be performed on both the wild-type and mutant cells and tissues where DmPKD2 is normally expressed. The long-term goal of this research is to define the biochemical pathway by which the PKD2 family of channels operates to regulate cell and developmental processes.

描述（由申请人提供）： 最近在理解常染色体显性方面取得了很多进展 多囊性肾脏疾病（ADPKD），会影响全球数百万的人。 主要疾病表型的特征是持续增殖和 肾上皮细胞的脱不同，最终形成流体填充的 囊肿。逐渐地，这些囊肿会导致解剖失真和肾衰竭。 引起疾病的基因之一PKD2具有显着的总体序列，并且 结构保护到各种已知的阳离子通道。最近的研究表明 PKD2或相关蛋白PKDL的表达足以产生 CA2+ - 可渗透的非选择性阳离子通道活动。另一种ADPKD疾病 基因PKD1也是一种跨膜蛋白，已被证明是相互作用的 与PKD2。 PKD 1可以调节启动的PKD2通道活动 保持正确的细胞内信号转导途径 上皮细胞极性，分泌和其他功能。该建议适用 新开发的功能基因组学工具通过 它在果蝇中的同源物，即DMPKD2基因。我们已经克隆了DMPKD2 基因及其与人类PKD2的相似性相似，在 两个最保守的区域，其中包含预测的跨膜段。 dmpkd2的n-和c-termini。更具分歧。我们在这里建议测试 假设DMPKD2充当通道，如其他脊椎动物所示 PKD2相关蛋白。 DMPKD2的属性和调节将是 量化。我们还建议测试DMPKD2在其中的通道函数 天然环境，即活蝇上皮组织。具体来说，我们会的 遗传过表达或消融培养细胞中DMPKD2的产生 （体外）和蝇组织（体内）。将进行单通道分析 在野生型和突变细胞和组织上，DMPKD2通常为 表达。这项研究的长期目标是定义生化 PKD2通道家族运行以调节细胞的途径 发展过程。