BIOCHEMICAL DIFFERENTIATION OF SOMATIC MOTOR NEURONS

体运动神经元的生化分化

• 批准号: 3328381
• 负责人: Arlene Y Chiu
• 金额: $ 14.28万
• 依托单位: BECKMAN RESEARCH INSTITUTE/CITY OF HOPE
• 依托单位国家: 美国
• 财政年份: 1990
• 资助国家: 美国
• 起止时间: 1990-01-03 至 1994-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3328381
• 关键词: Rodentias; antigens; autonomic nervous system; brain stem; cell differentiation; developmental neurobiology; hybridomas; immunocytochemistry; innervation; intermediate filaments; monoclonal antibody; motor neurons; neuroanatomy; neurogenesis; spinal cord; tissue /cell culture

The long term goal of this project is to understand the steps in the development and maturation of the motor neuronal phenotype, and how extrinsic factors influence this process. Toward this end, our current efforts have been focused on generating a panel of monoclonal antibodies with which to identify somatic motor neurons during rodent development. Once the sequential appearance of such markers, during development, is determined, the antigenic profile of a motor neuron will serve as an accurate means of staging its state of differentiation. We have now generated several antibodies that selectively recognize somatic motor neurons. One of these shows a very strict specificity for this class of neurons, and immunoreactivity appears late in development. In contrast, two other antibodies stain relatively immature motor neurons. These results demonstrate that molecular markers of motor neuronal differentiation exists, and that monoclonal antibody techniques can be successfully used to produce probes to such antigens. We will continue to expand our panel of markers for this class of neurons, using novel immunization protocols, in order to obtain a complete antigenic profile of motor neurons throughout development. With the antibodies that we have already generated, our first aim is to examine carefully the spatial and temporal distribution of immunoreactive epitopes, in different populations of somatic motor neurons in the developing brainstem and spinal cord. We will then perturb afferent inputs or target contact to ask if acquisition or maintenance of immunoreactivity is dependent on these parameters. We will also biochemically characterize and identify the immunoreactive molecules present in motor neurons. Finally, we propose to examine the differentiation of motor neurons in vitro using a spinal slice preparation. We will ask whether the antigens expressed by motor neurons in situ can be acquired in culture, and with the same specificity and time course. These experiments, in conjunction with studies in situ, will provide information on the molecular differentiation of this important class of cholinergic neurons.

该项目的长期目标是了解 运动神经元表型的发育和成熟，以及如何 外在因素影响这一过程。 为此，我们当前 工作重点是产生一组单克隆抗体 用于识别啮齿动物发育过程中的体运动神经元。 一旦这些标记在发育过程中连续出现， 确定后，运动神经元的抗原谱将作为 分期其分化状态的准确方法。 我们现在已经产生了几种选择性识别体细胞的抗体 运动神经元。 其中之一显示了该类的非常严格的特殊性 神经元的数量和免疫反应性出现在发育后期。 相比之下， 另外两种抗体对相对不成熟的运动神经元进行染色。 这些 结果表明，运动神经元的分子标记 差异化存在，单克隆抗体技术可以 成功地用于生产此类抗原的探针。 我们将继续 使用新颖的方法扩展此类神经元的标记物组 免疫方案，以获得完整的抗原谱 运动神经元贯穿整个发育过程。 利用我们已经产生的抗体，我们的首要目标是 仔细检查免疫反应的空间和时间分布 表位，在不同的体细胞运动神经元群体中 脑干和脊髓正在发育。 然后我们将扰乱传入输入 或目标接触，询问是否获得或维持免疫反应性 取决于这些参数。 我们还将进行生化表征 并识别运动神经元中存在的免疫反应分子。 最后，我们建议检查运动神经元的分化 体外使用脊髓切片制备。 我们会询问抗原是否 运动神经元原位表达的表达可以在培养物中获得，并且 相同的特异性和时间进程。 这些实验结合 原位研究，将提供分子分化的信息 这一类重要的胆碱能神经元。