NEURONS AND GLIA IN VITRO

体外神经元和神经胶质细胞

• 批准号: 3396358
• 负责人: BRUCE Robert RANSOM
• 金额: $ 17.4万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1987
• 资助国家: 美国
• 起止时间: 1987-09-01 至 1991-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3396358
• 关键词: acid base balance; astrocytes; cerebral cortex; cerebral ischemia /hypoxia; developmental neurobiology; electrophysiology; extracellular; glia; histology; homeostasis; laboratory mouse; laboratory rat; membrane permeability; microelectrodes; neural information processing; neuroanatomy; neurochemistry; neurons; neurophysiology; optic nerve; tissue /cell culture; voltage /patch clamp

The proposed research represents a continuation of studies, in vitro, of some of the basic physiological and morphological properties of mammalian glia and neurons, in relation to the ionic and volume regulation of the extracellular space (ECS) of brain. The long term goals of this work are to discover how the basic properties of glia and neurons interact to account for the dynamic, but regulated, ionic and volume flucuations seen in the ECS with brain activity. A new direction for this proposal is to study this issue in a pathological circumstance; specifically, to analyze the pathophysiology of anoxia in white matter of the brain, in mature and developing animals, using the isolated rat optic nerve (RON). Isolated RONs, tissue slices from rat cortex, and cultured astrocytes, will be used in most experiments and offer several advantages over in vivo preparations, including ready access of perfusion solutions to the ECS, direct visualization of microanatomic features of the tissue cells, and great experimental flexibility. Using standard electrophysiological recording techniques, ion-sensitive microelectrodes, and the patch-clamp method, the following projects will be undertaken: 1) developmental study of glial cell physiology and morphology in the RON; 2) patch-clamp studies of single ion channels in cultured glia; 3) analysis of activity dependent and K+ - induced changes in pHo in the RON and cortex; 4) studies on the pathophysiology of anoxia in white matter, using the RON; 5) studies on the mechanism(s) of neural activity dependent and K+ - induced ECS shrinkage; 6) developmental studies on the susceptibility of white matter to anoxic injury. These studies should provide basic new information about the properties of glia and neurons, especially as these relate to ion homeostasis in brain ECS under normal and pathological conditions. These studies are broadly relevant to the clinical problems of stroke, neonatal asphyxia, and brain swelling.

拟议的研究代表了研究的延续， 体外，一些基本的生理和形态学 哺乳动物神经胶质细胞和神经元的特性与离子的关系 和大脑细胞外空间（ECS）的体积调节。 这项工作的长期目标是发现基本的 神经胶质细胞和神经元的特性相互作用以解释动态， 但在 ECS 中观察到受调节的离子和体积波动 大脑活动。 该提案的一个新方向是研究这个 病理情况下的问题；具体来说，要分析的是 成熟大脑白质缺氧的病理生理学 和发育中的动物，使用离体的大鼠视神经（RON）。 分离 RON、大鼠皮层组织切片并进行培养 星形胶质细胞，将用于大多数实验并提供多种 与体内制剂相比的优势，包括易于获取 ECS 的灌注解决方案，直接可视化 组织细胞的显微解剖特征以及出色的实验 灵活性。 使用标准电生理记录 技术、离子敏感微电极和膜片钳 方法，将开展以下项目：1) 胶质细胞生理学和形态学的发育研究 罗恩； 2）培养物中单离子通道的膜片钳研究 神经胶质细胞; 3) 活动依赖性和 K+ 诱导的变化分析 RON 和皮质中的 pHo； 4）病理生理学研究 白质缺氧，使用 RON； 5）研究 神经活动依赖性和 K+ 诱导的 ECS 机制 收缩； 6）白人易感性的发育研究 与缺氧损伤有关。 这些研究应该提供基本的新 有关神经胶质细胞和神经元特性的信息，尤其是 因为这些与正常和正常情况下脑 ECS 中的离子稳态有关 病理状况。 这些研究与广泛相关 中风、新生儿窒息和脑肿胀的临床问题。