Role of CIC-5 Endocytosis and NHES Trafficking

CIC-5 胞吞作用和 NHES 贩运的作用

• 批准号: 8379307
• 负责人: Sandra Guggino
• 金额: $ 37.02万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-09-15 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/8379307
• 关键词: Abbreviations; Acids; Address; Adult; Affect; Albumins; Amino Acids; Antibodies; Antigens; Apical; Back; Basal Cell; Biotinylation; Brush Border; CLC Gene; Calcium; Cell Line; Cell membrane; Cells; Chloride Channels; Chloride Ion; Chlorides; Cleaved cell; Confocal Microscopy; Coupled; Data; Defect; Dexamethasone; Dextrans; Didelphidae; Dipeptidyl Peptidases; Disease; Disease Outcome; Early Endosome; Endocytosis; Endosomes; Epithelial Cells; Excretory function; Exhibits; Exocytosis; Fanconi Syndrome; Genes; Glucosephosphates; Glycosuria; Goals; Golgi Apparatus; Grant; Hereditary Disease; Hormones; Horseradish Peroxidase; Human; Instruction; Intestines; Kidney; Kidney Calculi; Kidney Failure; Knock-out; Knockout Mice; LDL-Receptor Related Protein 2; Label; Link; Liquid substance; Location; Lysosomes; Mannosidase; Measures; Mediating; Membrane; Membrane Protein Traffic; Membrane Transport Proteins; Metabolic; Microscopy; Modeling; Molecular; Molecular Weight; Movement; Mus; Mutate; Mutation; Na(+)-K(+)-Exchanging ATPase; Nephrocalcinosis; Nephrolithiasis; Nephrons; Normal Cell; Nutrient; Organelles; Pain; Parathyroid gland; Pathway interactions; Patients; Pattern; Phase; Phenotype; Physiologic pulse; Polyuria; Process; Property; Proteins; Proteinuria; Protons; Proximal Kidney Tubules; Radio; Radiolabeled; Rat-1; Recycling; Renal tubule structure; Retrieval; Role; Secondary to; Sodium; Subfamily lentivirinae; Surface; Syndrome; Testing; Transferrin; Vitamin D-Binding Protein; Water; Western Blotting; Wild Type Mouse; absorption; alanine aminopeptidase; apical membrane; base; basolateral membrane; brush border membrane; calcification; cellular microvillus; channel blockers; chromophore; dextran; disease phenotype; hypercalciuria; inhibitor/antagonist; inorganic phosphate; insight; kidney cell; kidney cortex; late endosome; male; membrane synthesis; mouse model; prevent; radiotracer; receptor; research study; residence; response; small hairpin RNA; sodium phosphate; sodium-phosphate cotransporter proteins; tetramethylrhodamine; trafficking; two-photon; urinary; voltage

Dent disease, is a disorder that is characterized by low molecular weight proteinuria, aminoaciduria, glycosuria, hyperphosphaturia (Fanconi syndrome) loss of sodium and water as well as hypercalciuria, nephrocalcinosis, and nephrolithiasis. Dent disease has been established to be caused by mutations of the CICN-5 gene, the product of which is CIC-5 voltage-gated Cl/H exchanger expressed in highest abundance in renal tubules ofthe proximal nephron. Our preliminary studies in a CICn-5 knockout mouse model, show that it recapitulates the features of Dent disease including low molecular weight proteinuria, aminoaciduria, glycosuria, loss of sodium, polyuria, hyperphosphaturia, hypercalciuria, renal calcifications, and renal failure. The proposed study will define the underlying molecular basis of these reabsorption defects. Our studies have shown CIC-5 Is necessary for exocytosis of NHE3 under basal and stimulated conditions. Specific aim 1. We hypothesize that loss of Clc-5 or an alkaline pH In the recycling endosome causes decreased exocytic trafficking of NHES and less surface NHES. Total NHES is not reduced because synthesis and surface delivery of NHES does not occur via the recycling endosome and is normal. Specific aim 2. We hypothesize that surface megalin in CICn-5 ko proximal tubules is decreased because megalin Is arrested in the recycling endosome when CIC-5 is lacking, or there is an alkaline pH in this compartment. Another option that may alter megalin at the surface is decreased surface NHES activity. We hypothesize that the loss of CIC-5, or increased endosomal pH causes less synthesis and apical delivery of megalin because this process occurs through the recycling endosome. Specific aim S. We hypothesize that the loss of CIC-5, or Increased endosomal pH causes less synthesis and delivery of newly synthesized Npt2a to the surface because this process occurs via a CIC-5 positive compartment. RELEVANCE (See instructions): This grant will outline the molecular mechanisms underlying the kidney defects in Dent disease. The outcome of this disease Is loss of important nutrients Including proteins, sodium and phosphate. This disease also causes painful kidney stones and kidney failure. In order to treat any of these outcomes of this disease we need to know more about how the faulty movement of key proteins are caused.

凹陷疾病是一种疾病，其特征是低分子量蛋白尿，氨基酸尿症， 糖尿，高磷酸（Fanconi综合征）钠和水的损失以及高钙尿症， 肾上腺素病和肾结石病。已经确定了疾病是由突变引起的 CICN-5基因，其乘积是CIC-5电压门控的Cl/H交换器，以最大的丰度表示 在近端肾单位的肾小管中。我们在CICN-5敲除鼠标模型中的初步研究，显示 它概括了凹痕疾病的特征，包括低分子量蛋白尿，氨基酸尿症， 糖尿，钠，多尿，高磷酸，高钙，肾脏钙化和肾衰竭的丧失。 拟议的研究将定义这些重吸收缺陷的基本分子基础。我们的研究 已经表明CIC-5对于基础和刺激条件下NHE3的胞吐作用是必需的。具体目标 1。我们假设回收内体中CLC-5或碱性pH的损失会导致外旋细胞减少 nhes的运输和较少的表面nhes。由于合成和表面 NHES的递送不会通过回收内体发生，并且是正常的。具体目标2。我们假设 CICN-5 KO近端小管中的表面巨林由于在回收中被捕而降低 当缺乏CIC-5时，内体或该隔室中有碱性pH值。可能的另一个选择 在表面上改变梅加林的表面活性降低。我们假设CIC-5或 内体pH的增加会导致较少的合成和梅加林的顶递送，因为此过程发生 通过回收的内体。具体目的S.我们假设CIC-5的丢失或增加 内体pH会导致较少的合成和新合成的NPT2A向表面的递送 过程通过CIC-5正隔室发生。 相关性（请参阅说明）： 该赠款将概述凹陷疾病中肾脏缺陷的分子机制。这 这种疾病的结果是蛋白质，钠和磷酸盐在内的重要营养素的丧失。这种疾病 还会导致疼​​痛的肾结石和肾衰竭。为了治疗这种疾病的任何结果 我们需要更多地了解关键蛋白的错误运动是如何引起的。