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Renal Effects of Aldosterone

醛固酮对肾脏的影响

基本信息

批准号：
10266000
负责人：
David H Ellison
金额：
--
依托单位：
PORTLAND VA MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-01-01 至 2022-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10266000
关键词：
Addison&apos s disease Address Adrenal Glands Aftercare Aldosterone Aldosterone Antagonists Binding Biological Markers Blood Pressure Brain CYP11B2 gene Candidate Disease Gene Cardiovascular Diseases Carrier Proteins Cell Nucleus Cells Consumption Diet Disease Distal Distal convoluted renal tubule structure Doxycycline Duct (organ) structure Ductal Epithelial Cell EFRAC Effectiveness Electrolytes Epithelial Epithelial Cells Excretory function Exhibits Funding Genes Genetic Genetic Transcription Glucocorticoids Goals Heart failure Homeostasis Hormones Human Hyperaldosteronism Hypertension Hypotension Infusion procedures KCNJ1 gene Kidney Knock-out Knockout Mice Knowledge Label Lead Life Measures Mediating Mediator of activation protein Metabolic Mineralocorticoid Receptor Mineralocorticoids Mus Nephrons Organ Pathway interactions Patients Perinatal mortality demographics Pharmaceutical Preparations Phenotype Phosphotransferases Physiology Plasma Population Potassium Potassium Channel Process Production Protein Analysis Proteins Receptor Activation Regulation Reporting Research Personnel Resistance Role SLC11A2 gene SLC12A3 gene Secondary Hypertension Serum Signal Transduction Site Sodium Sodium Channel Sodium Chloride Testing Therapeutic Tissues Transcript Translating Veterans Water Whole-Cell Recordings Work Zona Glomerulosa design dietary salt differential expression epithelial Na+ channel exosome experimental study fighting hyperkalemia in vivo novel novel strategies patch clamp receptor response salt intake side effect steroid hormone thiazide transcriptome transcriptome sequencing urinary wasting

项目摘要

The goal of this work is to determine how aldosterone causes cardiovascular disease, including hypertension and heart failure. Mice in which the mineralocorticoid receptor can be deleted in the kidney only will be used to examine aldosterone's role in signaling. These mice exhibit diminished sodium flux through both the thiazide-sensitive NaCl cotransporter and the epithelial sodium channel. Yet transport via the former is fully normalized when the mice are placed on low potassium diet, indicating that sodium channel stimulation is the major direct effect of aldosterone. The proposed experiments will test whether mineralocorticoid receptors along the second part of the distal convoluted tubule, known as the DCT2, modulate sodium chloride transport, but do so by occupancy with glucocorticoids, rather than by aldosterone. This will be done by comparing effects of deleting the aldosterone synthase gene, which leads to deficient aldosterone production, with the effects of deleting the mineralocorticoid receptor. The activity of the sodium channel will be measured using patch clamp and whole cell recording and the activity of the thiazide-sensitive NaCl cotransporter will be assessed using thiazide testing and phosphor-protein analysis. If channel transport activity is low in the receptor knockout, but not the synthase knockout, this will support the hypothesis. In a second aim, novel mediators of aldosterone signaling will be sought. We will dissect collecting ducts from mice with and without mineralocorticoid receptors, and consuming diets of varied electrolyte content. These collecting ducts will be subjected to RNAseq, to determine transcriptional differences that may underlie aldosterone effects. We will compare the transcripts differentially expressed in high aldosterone states versus those absent in mice with low aldosterone or lacking mineralocorticoid receptors. We will then select candidate genes to be tested using cultured epithelial cells, and ultimately in vivo. We will seek proteins that are essential for stimulating sodium channels under high aldosterone conditions.

这项工作的目标是确定醛固酮如何引起心血管疾病，包括高血压和心力衰竭。盐皮质激素受体可以在小鼠体内缺失，肾脏仅用于检查醛固酮在信号传导中的作用。这些老鼠表现出减少钠通量通过噻嗪敏感的NaCl协同转运蛋白和上皮细胞钠离子通道然而，当小鼠被置于低钾饮食，表明钠通道刺激是主要的直接影响，醛固酮拟议的实验将测试盐皮质激素受体是否沿沿着远曲小管的第二部分，称为DCT2，调节氯化钠运输，但这样做的占用糖皮质激素，而不是醛固酮。这将是通过比较删除醛固酮合酶基因的影响，这导致缺乏醛固酮的产生，具有删除盐皮质激素受体的作用。的活性将使用膜片钳和全细胞记录来测量钠通道，噻嗪敏感性NaCl协同转运蛋白的活性将使用噻嗪检测进行评估，磷蛋白分析如果在受体敲除中通道转运活性较低，合酶基因敲除这将支持我们的假设在第二个目标中，将寻求醛固酮信号传导的新介质。我们将解剖从有和没有盐皮质激素受体的小鼠收集导管，并消耗以下饮食：改变电解质含量。这些收集管将经受RNAseq，以确定转录差异可能是醛固酮效应的基础。我们会把记录在高醛固酮状态与低醛固酮状态小鼠中不存在的差异表达醛固酮或缺乏盐皮质激素受体。然后，我们将选择候选基因，使用培养的上皮细胞进行测试，并最终在体内进行测试。我们将寻找那些在高醛固酮条件下刺激钠通道所必需的。