PROMOTING AND DIRECTING PROCESS OUTGROWTH FROM ADULT NEURONS

促进和指导成年神经元的生长过程

• 批准号: 6338924
• 负责人: DAMIEN P KUFFLER
• 金额: $ 18.58万
• 依托单位: UNIVERSITY OF PUERTO RICO MED SCIENCES
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-05-01 至 2001-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6338924
• 关键词: Rana; Schwann cells; alternatives to animals in research; axon; bioassay; denervation; diffusion; electron microscopy; electrophysiology; growth cones; growth media; histochemistry /cytochemistry; innervation; macrophage; microelectrodes; microscopy; mixed tissue /cell culture; motor neurons; nervous system regeneration; neuronal guidance; neurotrophic factors; spinal ganglion; synapses; synaptogenesis; tissue /cell culture; video recording system

Central nervous system injury normally results in permanent neuronal deficits, while neuronal connections lost due to peripheral nervous system injury are normally re-established. To understand how regeneration in the central nervous system can be made possible and to promote better re-innervation in the peripheral nervous system, we must understand the basic mechanisms underlying the reestablishment of synaptic connections: what induces neurons to extend processes (trophic influences), how is growth of axons directed towards their targets (tropic influences), and how do axons recognize their targets (targets recognition cues)? Favorable preparations for studying these problems at the cellular and molecular levels are provided by neurons, muscle fibers and macrophages isolated from adult frogs and maintained in tissue culture. This laboratory has established methods for isolating adult motor and sensory neurons which survive for more than four weeks and extend processes. Culture have been established of pure populations of Schwann cells, macrophages and isolated intact adult skeletal muscle fibers. Another preparation will be used is for an in vivo bioassay. This proposal examines 3 aspects of nerve regeneration: (1) A study of the trophic influences of denervated peripheral nerve and macrophages on neuronal process outgrowth. (2) A study of the tropic influence of denervated nerve and macrophages on neuronal process outgrowth (3) Determination of whether motoneuron synapses form with specificity at original synaptic sites on cultured adult muscle fibers (an indicator of selective target recognition). Sensory and motor neurons will be cultured with Schwann cells, macrophages and muscle fibers and in the presence of known and putative trophic factors and the interactions and influences observed using physiological, electrophysiological and morphological methods. Techniques to be used include tissue culture, immunocytochemistry, morphometric analysis of process outgrowth and electron microscopy of contacts between neurons, macrophages and muscle fibers. Electrophysiological studies of synaptic transmission between neurons and muscle fibers will be carried out in collaboration with Project 50. Electrophysiological and morphological studies of the interactions between sensory/motor neurons and co-culture glial cells will be carried out in collaborations with Project 51.

中枢神经系统损伤通常会导致永久性神经元损伤 缺陷，而神经元连接由于周围神经而丢失 系统损伤通常会重新建立。 要了解如何 中枢神经系统的再生成为可能 促进周围神经系统更好的重新神经支配，我们必须 了解重建的基本机制 突触连接：什么诱导神经元延长过程（营养 影响），轴突的生长如何定向到其目标 （热带影响），以及轴突如何识别它们的目标（目标 识别线索）？ 研究这些问题的有利准备 在细胞和分子水平上由神经元、肌肉提供 从成年青蛙中分离并保存在组织中的纤维和巨噬细胞 文化。 该实验室已建立了分离成人的方法 运动和感觉神经元存活超过四个星期 扩展流程。 纯种人群的文化已经建立 雪旺细胞、巨噬细胞和分离的完整成人骨骼肌 纤维。 另一种制剂将用于体内生物测定。 该提案考察了神经再生的 3 个方面：(1) 去神经末梢神经和巨噬细胞对营养的影响 神经元过程的生长。 (2) 热带影响研究 去神经神经和巨噬细胞对神经元突起生长的影响 (3) 确定运动神经元突触是否形成具有特异性 培养的成体肌纤维上的原始突触位点（一个指标 选择性目标识别）。 感觉和运动神经元将 与雪旺细胞、巨噬细胞和肌纤维一起培养 已知和推定的营养因子的存在以及相互作用和 使用生理学、电生理学和 形态学方法。 使用的技术包括组织培养、 免疫细胞化学、过程生长的形态分析和 神经元、巨噬细胞和肌肉之间接触的电子显微镜 纤维。 突触传递的电生理学研究 神经元和肌肉纤维将与 项目 50. 电生理学和形态学研究 感觉/运动神经元与共培养神经胶质细胞之间的相互作用 将与Project 51合作进行。