WATER COMPARTMENTALIZATION IN ISCHEMIA

缺血时的水区室化

• 批准号: 6243844
• 负责人: CHARLES NICHOLSON
• 金额: $ 19万
• 依托单位: NEW YORK UNIVERSITY SCHOOL OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-09-20 至 1998-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6243844
• 关键词: age difference; body fluid osmolarity; brain cell; cell membrane; cell morphology; cell water; cerebral ischemia /hypoxia; extracellular; glutamate receptor; glutamate transporter; guinea pigs; intracellular transport; kainate; laboratory rat; light scattering; membrane activity; neocortex; osmosis; sectioning

The overall goal of this project is to understand how water shifts between extracellular and intracellular compartments during ischemia, and during changes in the brain microenvironment that mimic ischemic events. Basic experiments will be done to choose and characterize an ischemic slice model and to look at related protocols that cause water movement in both normal and gliotic slices. A set of collaborative investigations with other members of the Center will then capitalize on this basic research. During all experiments data will be gathered on light scattering properties of the tissue and [Na] - [K] content to see if they will provide useful indicators of water behavior. Finally the data will be used to model changes in the geometry the extracellular space. The majority of experiments will use slices from the neocortex of rats, with which we have experience under both normal and ischemic conditions. The slice permits the application of different media and drugs at precisely known concentrations. Water compartmentalization will be measured using a combination of the extracellular TAMA-method, based on ion-selective microelectrodes, and the weighing-drying method. Aim 1. After setting up a reliable ischemic slice model we will determine how water moves in the intracellular and extracellular compartments, determine the role of NaCl entry into cells, and assess the role of extracellular calcium. Aim 2. Elucidates which of the changes in the cellular microenvironment that occur during ischemia cause water movements. We shall use the normal slice and elevate [K+]c block Na-K transport, change osmolarity of bathing medium, add glutamate and NMDA and AMPA blockers. Aim 3. Some of the experiments will be repeated on slices containing predominantly glial cells. Aim 4. (With Dr. Margaret Rice ) Studies the distribution of water in the young adult and ag ed guinea pig brain under normal and ischemic conditions both with and without ascorbate in the bathing medium. Aim 5. (With Dr. Mitchell Chesler) Looks at the relationship between the putative Na+ - HCO3 co-transporter and water compartmentalization in the gliotic slice under normal conditions, ischemia and when Na-K transport is blocked. Aim 6. Measures the light scattering properties of the slice and analyzes the total Na and K content for many of the above experiments to determine if these measures permit a simplified approach to charting water movement between compartments. The accumulated data will be used to test models relating volume fraction and tortuosity in the extracellular space.

这个项目的总体目标是了解水是如何在 缺血期间和期间的细胞外和细胞内间隔 模拟缺血事件的大脑微环境的变化。基本信息 将进行实验以选择和表征缺血切片模型 并查看在正常情况下导致水运动的相关方案 和神经胶质切片。与其他机构合作进行的一系列调查 然后，该中心的成员将利用这一基础研究。在.期间 所有实验数据都将收集关于光散射特性的 组织和[Na]-[K]含量，看看它们是否会提供有用的指示剂 水的行为。最后，数据将用于对 对细胞外空间进行几何学处理。 大多数实验将使用大鼠新皮质的切片， 这是我们在正常和缺血条件下都有的经验。 这种薄片可以精确地使用不同的介质和药物 已知浓度。水的分区将使用 基于离子选择性的胞外TAMA法的结合 微电极和称重-干燥方法。 目的1.建立可靠的脑缺血切片模型后，我们将确定 水在细胞内和细胞外的隔间里是如何运动的， 确定氯化钠进入细胞的作用，并评估 细胞外钙。目标2。阐明以下哪些变化 在缺血期间发生的细胞微环境会引起水的运动。 我们将使用正常切片，并提高[K+]c阻断Na-K转运， 改变沐浴介质渗透压，添加谷氨酸、NMDA和AMPA 拦截者。目标3.一些实验将在切片上重复 含有以胶质细胞为主的。《目标4》(与玛格丽特·赖斯博士合作) 水在幼年豚鼠和老龄豚鼠体内的分布 有无抗坏血酸的正常和缺血条件下的脑 在洗澡介质里。目标5。(与米切尔·切斯勒博士)看着 可能的Na+-HCO3共转运体与水的关系 正常情况下脑缺血时胶质细胞切片的区划 当钠钾转运受阻时。目标6.测量光散射 切片的性质，并分析了许多 上述实验以确定这些措施是否允许简化 绘制舱室间水流运动图的方法。累积的 数据将被用来测试与体积分数和弯曲度有关的模型 细胞外空间。