STUDIES ON MAMMALIAN NEURONS AND GLIA IN VITRO

拟议中的研究是对一些 基本生理、形态和药理学特性 哺乳动物神经胶质和神经元的体外系统。 这项研究也将 研究离子和 细胞外间隙（ECS）的体积波动 准备工作 其中一些问题将在不同的 在出生后的发展阶段，并与重要的 在这些时间发生的形态学或生物化学变化。 一个新 本计划的研究方向是单离子通道的分析 在培养或分离的神经胶质细胞中。 离体大鼠视神经，组织 来自大鼠新皮层的切片和培养的啮齿动物星形胶质细胞将用于 大多数实验，并提供了几个优势，在体内制备 包括灌注溶液到ECS的容易进入，直接 组织细胞的显微解剖学特征的可视化，以及相关的 神经元和神经胶质细胞的细胞内记录的简易性。 的一个组成部分 该项目将调查其中一些问题的特点， 非哺乳动物标本龟脑 使用细胞内染色 将开展以下项目：1） 大鼠神经胶质细胞生理和形态的发育研究 视神经; 2）电生理、形态学和代谢研究 慢电位产生机制 和ECS离子稳态; 4）表征神经元的 大鼠视神经和新皮质组织pH值的活动依赖性变化 切片; 5）神经活动依赖的ECS收缩机制; 6）补丁 神经胶质细胞离子通道的钳夹研究

The proposed research represents a continuation of studies on some of the basic physiological, morphological, and pharmacological properties of mammalian glia and neurons in in vitro systems. This research will also investigate the characteristics and homeostatic regulation of ionic and volume fluctuations in the extracellular space (ECS) of in vitro preparations. Some of these issues will be carefully studied at different stages during postnatal development and correlated with important morphological or biochemical changes occurring at these times. A new research direction for this plan is the analysis of the single ion channel in cultured or isolated glial cells. Isolated rat optic nerves, tissue slices from rat neocortex, and cultured rodent 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 relative ease of intracellular recordings from neurons and glia. One component of the project will investigate characteristics of some of these issues in a non-mammalian preparation, the turtle brain. Using intracellular staining and recording techniques the following projects will be undertaken: 1) developmental study of glial cell physiology and morphology in the rat optic nerve; 2) electrophysiological, morphological, and metabolic studies on glia in the rat optic nerve; 3) mechanisms of slow potential generation and ECS ionic homeostasis in turtle cortex; 4) characterization of neural activity-dependent changes of pHo in rat optic nerve and neocortical tissue slice; 5) mechanisms of neural activity dependent ECS shrinkage; 6) patch clamp studies on single ionic channels of glia in vitro.