GROWTH-ASSOCIATED PROTEINS AND NEURONAL REGENERATION

生长相关蛋白质和神经元再生

• 批准号: 3398153
• 负责人: JOHN A FREEMAN
• 金额: $ 16.87万
• 依托单位: VANDERBILT UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1982
• 资助国家: 美国
• 起止时间: 1982-07-01 至 1990-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3398153
• 关键词: active transport; axon; cell free system; central nervous system; complementary DNA; cyclic GMP; electrofocusing; electron microscopy; evolution; gel electrophoresis; genetic transcription; genetic translation; glia; histochemistry /cytochemistry; immunochemistry; immunologic assay /test; immunoprecipitation; laboratory rabbit; laboratory rat; messenger RNA; molecular weight; nervous system regeneration; neurogenesis; neurons; neurotrophic factors; nucleic acid hybridization; nucleic acid probes; protein biosynthesis; radiotracer; synapses

Recent work has led to the discovery of a class of growth-associated proteins, called GAPs, whose expression is selectively enhanced in regenerating and developing neurons of some species. The purpose of this investigation is to study the cellular role of these growth-associated proteins, and in particular to determine whether tje abo;otu pf a meirpm tp regenerate depends on its ability to induce their synthesis. There are five related objectives. The first objective is to determine, by means of a limited phylogenetic survey, whether these proteins appear in CNS neurons lacking the capability to regenerate, or whether they occur only in growing or regenerating neurons. Two very sensitive and powerful techniques will be employed to detect and quantitate these proteins: computer-analyzed 2-dimensional gel electrophoresis, and immunological techniques. The second objective is to determine the cellular localization of GAPs, using EM immunocytochemical localization methods. The third objective is to characterize the major distinguishing characteristics of GAPs, and the fourth is to investigate the cellular functions of GAPs, by correlating the kinetics of GAP expression with different phases of axonal growth in developing and regenerating systems. The last objective is to determine how the expression of GAPs is regulated. We plan to study the effects of several likely regulatory molecules on GAP synthesis, including nerve growth factor (NGF) and cyclic AMP. The site of regulation (transcriptional, translational, of post- translational) will be determined with inhibitors of RNA and protein synthesis. Finally, specific changes in GAP messenger RNA during regeneration will be observed using a cell-free synthesis system. The significance of these studies lies in their promise to reveal some molecular mechanisms which control axonal growth. These are of paramount importance in understanding the development of the nervous system, and its response to injury.

最近的研究发现了一类与生长相关的 被称为GAP的蛋白质，其表达在 某些物种的神经元再生和发育。这样做的目的是 调查是为了研究这些与生长相关的细胞作用 蛋白质，特别是确定是否有一种新的蛋白质 再生依赖于它诱导它们合成的能力。确实有 五个相关目标。第一个目标是通过以下方式确定 一项有限的系统发育调查，这些蛋白是否出现在中枢神经系统神经元中 缺乏再生的能力，或者是否只在生长过程中出现 或者再生神经元。两种非常敏感和强大的技术将 被用来检测和定量这些蛋白质：计算机分析 双向凝胶电泳法和免疫学技术。这个 第二个目标是确定间隙的细胞定位，使用 EM免疫细胞化学定位方法。第三个目标是 描述差距的主要区别特征，以及 第四是通过关联GAP的细胞功能来研究GAP 轴突生长不同阶段中GAP表达的动态变化 开发和再生系统。最后一个目标是确定 间隙的表达是如何被调控的。我们计划研究 几个可能调节GAP合成的分子，包括神经 生长因子(NGF)和环磷酸腺苷(CAMP)。监管的场所 (转录、翻译或翻译后)将确定 核糖核酸和蛋白质合成的抑制剂。最后，中的具体变化 在再生过程中将观察到GAP信使RNA使用无细胞 综合系统。这些研究的意义在于它们的前景 揭示控制轴突生长的一些分子机制。这些是 对于理解神经质的发育至关重要 系统及其对伤害的反应。