GROWTH-ASSOCIATED PROTEINS AND NEURONAL REGENERATION

生长相关蛋白质和神经元再生

• 批准号: 3398152
• 负责人: JOHN A FREEMAN
• 金额: $ 16.32万
• 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 1989-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3398152
• 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; messenger RNA; molecular weight; nervous system regeneration; neurons; neurotrophic factors; 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 蛋白质，称为GAP，其表达选择性增强， 某些物种的神经元再生和发育。 这样做的目的 研究的目的是研究这些生长相关的细胞作用， 蛋白质，并且特别是确定是否存在蛋白质， 再生取决于它诱导它们合成的能力。 有 五个相关目标。 第一个目标是确定，通过 一个有限的系统发育调查，这些蛋白质是否出现在中枢神经系统神经元 缺乏再生能力，或者它们是否只发生在生长中， 或再生神经元。 两个非常敏感和强大的技术将 用于检测和定量这些蛋白质：计算机分析 2-三维凝胶电泳和免疫学技术。 的 第二个目标是确定GAP的蜂窝定位，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， 第四是研究GAP的细胞功能， GAP表达的动力学与轴突生长的不同阶段， 开发和再生系统。 最后一个目标是确定 GAP的表达是如何调节的。 我们计划研究 几种可能的GAP合成调控分子，包括神经 生长因子（NGF）和环AMP。 调节部位 （转录的、翻译的、翻译后的）将被确定 RNA和蛋白质合成的抑制剂。 最后，具体变化 再生期间的GAP信使RNA将使用无细胞的 合成系统 这些研究的意义在于它们的承诺 以揭示一些控制轴突生长的分子机制。 这些是 对理解神经系统的发展至关重要 系统及其对损伤的反应。