REGENERATIVE PHENOMENA IN THE DEVELOPING ORGAN OF CORTI

柯蒂氏器官发育中的再生现象

• 批准号: 3217559
• 负责人: HANNA M SOBKOWICZ
• 金额: $ 16.78万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 1992
• 资助国家: 美国
• 起止时间: 1992-07-01 至 1996-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3217559
• 关键词: cell cycle; developmental neurobiology; ear hair cell; electron microscopy; epidermal growth factor; fibroblast growth factor; immunocytochemistry; injury; laboratory mouse; nervous system regeneration; neural degeneration; neurotrophic factors; newborn animals; organ of Corti; synaptogenesis; tissue /cell culture

The long-term goal of the proposed research is to gain understanding of the process of regeneration of sensory and nerve cells following mechanical injury to the cochlea in culture, and to define the factors that lead to their morphological recovery. In light of the lack of overt evidence proving that the mammalian organ of Corti is able to form new sensory cells, and given the fact that these cells are few and may be formed only once in a life span the significance of the research lies in understanding the mechanism of survival and recovery of receptor cells after injury. Specifically, we propose to study: 1) Post-traumatic cell proliferation in the organ of Corti: A) Mitotic cell proliferation in the injured cultures will be examined using 3H thymidine to investigate the identity of dividing cells, the duration of the mitotic response and the age- related occurrence of cell division; B) Nonmitotic cell proliferation, expressed by extensive sprouting of supporting cells, will be investigated in search of possible diffusable growth factors (NGF, EGF, FGF) released by the cells in response to direct injury or to the injury of sensory cells. 2) Regeneration of the injured sensory cells, especially the reformation of their cuticular plates and stereocilia. The study will encompass the morphological sequence of the regenerative phenomena, its comparison with normal development, and the influence of age on the regenerative capacity of sensory cells. 3) Degeneration and regeneration of spiral neurons, sprouting of their endings (using antibodies to GAP-43 and NF proteins), and formation of new ribbon synapses. The study will be done using short- and long-term cultures of the mouse organ of Corti. The injury will be done by hand, using either a pulled- glass pipettes or a laser. Light and electron microscopy, autoradiography, biochemistry and immunocytochemistry will be employed. Some specialized immunological assays will be done in collaboration with Dr. Philippe Lefebvre, University de Liege, Belgium.

拟议研究的长期目标是了解 随后的感觉和神经细胞再生的过程 培养中耳蜗的机械损伤，并定义因素 这导致了他们的形态恢复。鉴于缺乏公开 证据证明Corti的哺乳动物器官能够形成新的 感觉细胞，鉴于这些细胞很少，可能是 在生命中仅形成一次研究的重要性在于 了解生存和恢复受体细胞的机制 受伤后。 具体而言，我们建议研究：1）创伤后细胞增殖 在Corti的器官中：a）受伤的有丝分裂细胞增殖 将使用3H胸苷来研究培养物以研究身份 分裂细胞，有丝分裂反应的持续时间和年龄 细胞分裂的相关发生； b）非霉菌细胞增殖， 通过广泛的支持细胞发芽表达的将是 研究以寻找可能的扩散生长因子（NGF，EGF， FGF）由细胞释放，以响应直接损伤或损伤 感觉细胞。 2）受伤的感觉细胞的再生， 尤其是其表皮板和立体猫的改革。 该研究将涵盖再生的形态序列 现象，与正常发展的比较以及 年龄在感觉细胞的再生能力上。 3）变性和 螺旋神经元的再生，结尾的发芽（使用 GAP-43和NF蛋白的抗体）以及新功能区的形成 突触。 该研究将使用小鼠的短期和长期培养 Corti器官。受伤将使用拉动 - 玻璃管或激光。光与电子显微镜， 将采用放射自显影，生物化学和免疫细胞化学。 一些专门的免疫学测定将与 菲利普·勒费维（Philippe Lefebvre）博士，比利时列日大学。