DEVELOPMENT OF HIPPOCAMPAL NEURONS IN CULTURE

培养中海马神经元的发育

• 批准号: 2263143
• 负责人: GARY A BANKER
• 金额: $ 24.14万
• 依托单位: UNIVERSITY OF VIRGINIA
• 依托单位国家: 美国
• 财政年份: 1981
• 资助国家: 美国
• 起止时间: 1981-08-01 至 1997-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2263143
• 关键词: Golgi apparatus; antibody receptor; cellular polarity; chimeric proteins; dendrites; electron microscopy; exocytosis; fluorescence microscopy; growth cones; hippocampus; intracellular transport; laboratory rat; membrane proteins; molecular cloning; neural cell adhesion molecules; neural plasticity; neurogenesis; neuronal transport; tissue /cell culture; transferrin receptor; vesicle /vacuole; video microscopy

The long-term goal of this research is to understand from a cell biological perspective how neurons establish and maintain distinct axonal and dendritic domains that differ in structure and function. This feature of nerve cells -- referred to as neuronal polarity -- is essential to normal neural function and a disruption of neuronal polarity is thought to contribute to the pathophysiology of many neurologic diseases. An understanding of the cellular mechanisms underlying neuronal polarity will be necessary for the development of molecular therapies for neurologic diseases. The development and maintenance of polarity will be studied in cell cultures of embryonic rat hippocampal neurons, using techniques that range from observation and manipulation of living cells to investigations at the cellular and molecular levels. The particular questions to be addressed derive from previous work that has identified the key stages in the development of hippocampal neurons that determine their polarity. The initial emergence of the axon, the defining event in the establishment of polarity, will be analyzed by high resolution time-lapse microscopy under normal and experimental conditions. The response of more mature cells to axonal transection will be used to determine when polarity becomes irreversibly specified. Dendritic growth and the behavior of dendritic growth cones will be analyzed by time-lapse video microscopy. Electron microscopy will be used to identify ultrastructural features that distinguish dendritic from axonal growth cones. Sorting signals that direct integral membrane proteins to axons or dendrites will be identified. Defective Herpes viral vectors will be used to introduce cDNA constructs into cultured cells in order to assess how deletions in the endo-and ectodomains of polarized membrane proteins affect their distribution. Exocytic vesicle traffic between the Golgi complex and sites of incorporation into the plasma membrane will be analyzed at each stage of neuronal development using fluorescent lipid analogs.

这项研究的长期目标是从细胞中了解 生物学的观点神经元如何建立和保持独特的轴突 以及在结构和功能上不同的树突状域。 这 神经细胞的特征 - 称为神经元极性 - 正常神经功能和神经元极性的破坏至关重要 被认为有助于许多神经系统的病理生理学 疾病。 对基础机制的理解 神经元极性对于分子的发展是必要的 神经系统疾病的疗法。 的发展和维护 极性将在胚胎大鼠海马的细胞培养物中进行研究 神经元，使用从观察和操纵的技术 在细胞和分子水平上进行研究的活细胞。 要解决的特定问题来自以前的工作 已经确定了海马神经元发展的关键阶段 这决定了它们的极性。 轴突的最初出现， 在建立极性的情况下定义事件将通过高 分辨率在正常和实验下的延时显微镜 状况。 更成熟的细胞对轴突横断的反应将 用于确定何时不可逆转地指定极性。 树突状的生长和树突生长锥的行为将是 通过延时视频显微镜分析。 电子显微镜将是 用于识别区分树突与树突的超微结构特征 轴突生长锥。 排序直接整体膜的信号 将确定蛋白质到轴突或树突。 有缺陷的疱疹 病毒载体将用于将cDNA构建体引入培养 细胞以评估如何在内部和外生域中的删除 极化膜蛋白会影响其分布。 外囊囊泡 高尔基体综合体与融合地点之间的交通 质膜将在神经元发育的每个阶段进行分析 使用荧光脂质类似物。