Laboratory of Integrated Kidney Function

总结 综合肾功能核心的目标是提供广泛的服务，并支持 计划项目组每个PI的研究项目。分子遗传学的进步极大地 拓宽了我们对肾脏生理和病理生理的理解。转基因动物提供 优雅的模型来检查单个蛋白质对维持正常功能的贡献， 疾病的病理生理学转基因动物被广泛用作一种强大的工具， 表征特定蛋白质、通道、泵或转运蛋白的生理或病理生理作用。 这些研究对于了解疾病的机制和寻找新的靶点具有重要意义。 临床疾病的治疗。然而，由于技术上的困难，敲除和 转基因小鼠模型主要限于血液和尿液电解质测量。我们能够 在体外和体内检查完整的肾功能，从整个动物到单个肾单位，通过肾 清除率分析和来自大鼠或小鼠的肾小管的微灌注。使用这些方法，我们 能够测量盐、水和酸碱转运以及肾脏中的蛋白质吸收。比较 在正常动物与特定蛋白质、通道或转运蛋白发生改变的转基因动物之间 将提供生理学、病理生理学和疾病适应方面的宝贵信息。集成 Kidney Function Core在肾脏生理学研究中发挥了独特的重要作用， 研究项目在计划项目补助金。这个核心是由训练有素的人谁是熟练的所有 评估从整个动物到单个肾单位水平的肾转运功能所需的技术。 该核心实验室提供的服务和协作为项目中的每个PI提供了工具， 将使他们能够将细胞和分子观察与生理和病理生理学联系起来， 流程.为了促进计划项目的总体目标，我们计划提供以下具体内容 项目中提出的服务：1。检测和比较Na+、K+、Ca 2+和Mg 2+的表型 在ROMK和WNK 4（PHAII）突变小鼠中观察到的转运至从KLHL 3和CUL 3突变小鼠获得的转运 小鼠（项目#1）; 2.在低浓度和高浓度条件下， 肾小管流速，以研究NKCC 1在调节集合小管K+转运中的生理作用 （项目#2）; 3.检查生理抑制（通过PTH，多巴胺）或刺激（通过Ang II， 糖皮质激素）的NHE 3活性，以响应NHE 3磷酸化的位点， 减少NHE 3活性（项目#3）;和4.目的：研究缺血性脑损伤对电解质转运的影响， 显示AMPK表达改变的动物模型（项目#4）。

Summary The goal of the Integrated Kidney Function Core is to provide a wide range of services and to support the research projects of each PI of the Program Project Group. The progress in molecular genetics has greatly broadened our understanding of renal physiology and pathophysiology. Genetically altered animals provide elegant models to examine the contributions of individual proteins to the maintenance of normal function and pathophysiology of disease. Genetically modified animals are widely used as a powerful tools with which to characterize the physiological or pathphysiological roles of specific proteins, channels, pumps or transporters. These studies are important to understand the mechanisms of diseases and to look for novel targets for the treatment of clinical diseases. However, due to technical difficulties the characterization of knockout and transgenic mouse models has been largely limited to blood and urine electrolyte measurements. We are able to examine complete renal functions in vitro and in vivo, from whole animals to single nephrons, by renal clearance analysis and by microperfusion of renal tubules from rat or mouse. Using these methods, we are able to measure salt, water and acid-base transport as well as protein absorption in the kidney. Comparisons between normal vs. genetically modified animals in which a specific protein, channel or transporter is altered will provide invaluable information on physiology, pathophysiology and adaptation to diseases. The Integrated Kidney Function Core has played a uniquely important role in renal physiology research by supporting many research projects in the Program Project Grant. This core is staffed by well trained people who are skilled in all techniques needed to assess renal transport functions from the whole animal to the single nephron levels. Services and collaborations provided from this core lab provide each PI in the program project with tools that will allow them to connect their cellular and molecular observations to physiological and pathophysiological processes. To facilitate the overall goal of the Program Project, we plan to provide the following specific services as proposed in the projects: 1. To examine and compare the phenotypes of Na+, K+ Ca2+ and Mg2+ transport observed in ROMK and WNK4 (PHAII) mutant mice to those obtained from KLHL3 and CUL3 mutant mice (Project #1); 2. To measure both net Cl- transport and cellular Cl- concentrations under low and high tubular flow rates to study the physiological role of NKCC1 in the regulation of collecting tubule K+ transport (Project # 2); 3. To examine physiological inhibition (By PTH, dopamine) or stimulation (by Ang II, glucocorticoid) of NHE3 activity in response to NHE3 phosphorylation at sites that correlate with increased or reduced NHE3 activities (Project #3); and 4. To study electrolyte transport and the effects of ischemic injury in animal models manifesting altered AMPK expression (Project #4).