ION TRANSPORT IN COLLECTING CELL CULTURES

收集细胞培养物中的离子传输

• 批准号: 3238160
• 负责人: ELSA N. BELLO-REUSS
• 金额: $ 15.12万
• 依托单位: UNIVERSITY OF TEXAS MED BR GALVESTON
• 依托单位国家: 美国
• 财政年份: 1986
• 资助国家: 美国
• 起止时间: 1986-08-01 至 1993-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3238160
• 关键词: aldosterone; basolateral membrane; cell type; dopamine; electrophysiology; epithelium; hormone regulation /control mechanism; ion transport; kidney cell; laboratory rabbit; membrane permeability; membrane potentials; microelectrodes; mixed tissue /cell culture; neurotransmitters; norepinephrine; potassium; renal tubule acidosis

We aim to understand the ion transport properties and the mechanisms of its regulation in the different cell types of the cortical collecting duct (CCD). Specific aims are: 1. Continue characterizing the ion-transport mechanisms expressed in CCD cells in culture. Mixed CCD cultures will be used to identify mechanisms of ion transport, and the effects of ADH, aldosterone, and isoproterenol on the different transporters. 2. To study the effects of intra and extracellular pH on the cell biology and transport functions of CCD monolayers. 2.1. To assess the modifications in the proportion of the different cell types and the morphological changes in response to alterations in acid-base status of the cultures. 2.2. To establish the pH dependence of the transporters located in the apical and basolateral membranes of the different cells. We will examine the conductive and nonconductive ion-transport pathways of the three cell types under basal conditions and their responses to acute and chronic changes in pH of basolateral and/or apical solutions. 3. To characterize, structurally and functionally, homogeneous monolayers of principal cells and intercalated cells. 3.1. The ultrastructure and function of principal- cell monolayers will be studied: We will characterize at the cell-membrane level the baseline ion transport mechanisms and their regulation by hormones (ADH, aldosterone, isoproterenol). 3.2. Homogeneous cultures of intercalated cells, initially identified as either alpha or beta cells, will be studied with and without hormonal stimulation. Special consideration will be given to the possibility of transformation of a cell type to another. Monolayers of CCD cells will allow us to gather basic information on ion transport mechanisms at the cellular and single-membrane levels. They should also be useful tools for biochemical and molecular studies. Homogenous cultures will permit quantitative studies without the confounding influences of neighboring cells of different type. These studies could help our understanding of the pathogenesis of renal tubule ion-transport defects, such as renal tubular acidosis, concentration defects and K+ secretion abnormalities.

我们旨在了解离子传输属性及其机制 皮质收集管的不同细胞类型的调节 （CCD）。 具体目的是：1。继续表征离子传输 在培养中CCD细胞中表达的机制。 混合的CCD文化将是 用于鉴定离子传输机制和ADH的影响， 醛固酮和异丙醇在不同的转运蛋白上。 2。学习 细胞内pH和细胞外pH的影响对细胞生物学和运输的影响 CCD单层的功能。 2.1。评估修改 不同细胞类型的比例和形态变化 对培养物酸碱状态改变的反应。 2.2。到 建立位于顶端的转运蛋白的pH依赖性 不同细胞的基底外侧膜。 我们将检查 三种细胞类型的导电和非导电离子传输途径 在基础条件下及其对急性和慢性变化的反应 基底外侧和/或顶端溶液的pH。 3。要表征， 在结构和功能上，主要细胞的均匀单层 和插入的细胞。 3.1。主结构和主的功能 - 将研究细胞单层：我们将在细胞膜上进行表征 将基线离子传输机制及其调节 激素（ADH，醛固酮，异丙肾上腺素）。 3.2。同质文化 插入的细胞，最初被鉴定为α或β细胞， 将在有和没有激素刺激的情况下进行研究。 特别的 考虑到细胞转换的可能性 输入另一个。 CCD细胞的单层将使我们能够收集基本 有关细胞和单个膜处的离子传输机制的信息 水平。 它们还应该是生化和分子的有用工具 研究。 同质文化将允许无需定量研究 混淆不同类型的相邻细胞的影响。 这些 研究可以帮助我们理解肾小管的发病机理 离子传输缺陷，例如肾小管酸中毒，浓度 缺陷和K+分泌异常。