Isolation and characterization of rat kidney active urea transporter

大鼠肾活性尿素转运蛋白的分离和表征

基本信息

批准号：
7992617
负责人：
Guangping Chen
金额：
$ 10万
依托单位：
EMORY UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-12-15 至 2010-11-30
项目状态：
已结题

项目摘要

DESCRIPTION (provided by applicant): Urea plays an important role in the urinary concentrating mechanisms. In the past decade, significant progress has been made in understanding urea reabsorption and recycling in kidney including the cloning of the two facilitated urea transporter genes, UT-A and UT-B. However, strong evidence from physiological studies demonstrated that the existence of a sodium- dependent active transporter in mammalian kidney, which can uphill move urea in some nephron segments. Though the active urea transporter was proposed over 40 years ago, none has yet been characterized. The overall objective of this project is to clone this novel active urea transporter from kidney inner medulla (IM) by using two independent approaches: 1) modified two-tester suppression subtraction hybridization (ttSSH) and 2) expression screening using a urea uptake defective yeast strain. After having successfully identified the novel urea transporter, we will investigate the nature of the active urea transporter. We hope the two approaches we apply here will lead us to discover the new genes. We believe the success of this project will fill in the gap with the molecular evidence how urea is actively transported in kidney and will extend our understanding on the urinary concentrating mechanism and water homeostasis. PUBLIC HEALTH RELEVANCE Strong physiological evidence demonstrated the existence of active urea transport activity in mammalian kidney over 40 years. The overall objective of this project is to clone the novel active urea transporter from kidney IM by using two independent approaches. The success of this project will extend our understanding on the urinary concentrating mechanism and water homeostasis.

描述（由申请人提供）：尿素在尿液浓缩机制中起重要作用。在过去的十年中，在理解肾脏的重吸收和回收方面取得了重大进展，包括两个促进的尿素转运蛋白转运蛋白基因UT-A和UT-B的克隆。但是，来自生理研究的强有力证据表明，哺乳动物肾脏中钠依赖性活性转运蛋白的存在，这可以使某些肾单位段中的尿素移动尿素。尽管活跃的尿素运输蛋白是在40年前提出的，但尚未表征。该项目的总体目的是通过使用两种独立的方法克隆从肾脏内髓质（IM）中克隆这种新型的活性尿素转运蛋白：1）使用尿素吸收有缺陷的酵母菌菌株进行了修改后的两核抑制减法杂交（TTSSH）和2）表达筛查。成功鉴定出新型尿素转运蛋白后，我们将研究活性尿素转运蛋白的性质。我们希望我们在这里采用的两种方法将导致我们发现新基因。我们认为，该项目的成功将通过分子证据来填补空白，尿素是如何在肾脏中积极运输的，并将扩展我们对尿液浓缩机制和水稳态的理解。公共卫生相关性有力的生理证据表明，在40年内，哺乳动物肾脏中存在主动尿素转运活性。该项目的总体目的是使用两种独立方法克隆肾脏IM的新型活性尿素转运蛋白。该项目的成功将扩展我们对尿液集中机制和水稳态的理解。