NERVE GROWTH FACTOR SIGNALLING

神经生长因子信号传导

• 批准号: 2460594
• 负责人: MARIE W WOOTEN
• 金额: $ 11.93万
• 依托单位: AUBURN UNIVERSITY AT AUBURN
• 依托单位国家: 美国
• 财政年份: 1995
• 资助国家: 美国
• 起止时间: 1995-08-01 至 1999-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2460594
• 关键词: PC12 cells; cell cycle proteins; cyclic AMP; enzyme activity; guanine nucleotide binding protein; immunoprecipitation; isozymes; mitogen activated protein kinase; neurogenesis; neurotrophic factors; phosphoproteins; protein kinase C; transfection

Protein kinase C (PKC) has been implicated as a component of the Nerve growth factors (NGF) signalling pathway in PC12 cells; although, disagreement regarding the role that PKC plays in this process has persisted in the literature. We provide novel information demonstrating that activation of atypical zeta-PKC relates to the signalling paradox surrounding NGF and PKC isoforms. In addition, we also document that zeta- PKC is activated by NGF in a trk-dependent manner and modulated in a positive fashion by increases in cAMP. We hypothesize that zeta-PKC lies in parallel with raf kinases and is a major constituent of a newly described signalling pathway serving as a MAP kinase kinase kinase (MEK kinase). The goal of this project is to precisely define the role which zeta-PKC plays in mediating signal-coupling and neurite outgrowth. In order to fulfill this goal, two specific aims will be undertaken. We are proposing to; 10 establish the means by which activation of zeta-PKC modulates an existing or newly defined pathway by studying its interaction with proteins which comprise athe classical NGF signalling cascade involving ras / raf yieldsMAP kinase and 2) overexpress zeta-PKC to upregulate its signal and study both biochemical and morphological aspects of overexpression. The long-term goal of these studies is to establish the mechanism whereby zeta-PKC regulates NGF dependent and independent signalling involving neuronal differentiation. To accomplish these aims we will employ PC12 cells deficient in zeta-PKC or those overexpressing this isoform. The regulation exerted by zeta-PKC will be studied using a range of techniques: in situ activity assays, immunoprecipitation of individual signaling components, reconstitution, protein purification, peptide mapping and phosphoamino acid analysis. The outcome of these studies stand to change our conceptual framework of NGF-signal coupling and neurite outgrowth. Moreover, the findings generated by this study will have a direct impact upon Alzheimer's research, since PC12 cells have been used as a model system to elucidate the role of amyloid precursor protein (APP). Previous work in PC12 cells has demonstrated that APP is capable of enhancing neurite outgrowth and acts as a mediator of NGF's effects. Hence, elucidating the signalling pathway(s) leading to neurite extension will bear directly into the etiology and treatment of Alzheimer's, as well as, other human disease in which alterations in PKC and NGF are hypothesized to play a role.

蛋白激酶C（PKC）已与神经的成分有关 PC12细胞中的生长因子（NGF）信号通路；虽然， 关于PKC在此过程中所起的作用的分歧 在文学中坚持下去。 我们提供新的信息证明 非典型Zeta-PKC的激活与信号悖论有关 周围NGF和PKC同工型。 此外，我们还记录了Zeta- PKC被NGF以TRK依赖性方式激活，并在A中调制 积极的时尚营地增加。 我们假设Zeta-PKC在 与RAF激酶并行，是新的主要组成部分 描述了用作MAP激酶激酶激酶的信号通路（MEK 激酶）。 该项目的目的是精确定义角色 Zeta-PKC在介导信号偶联和神经突产物中发挥作用。 在 为了实现这一目标，将实现两个具体的目标。 我们是 提议10建立激活zeta-pkc的平均值 通过研究其相互作用来调节现有或新定义的途径 蛋白质包括A含有经典的NGF信号级联 涉及RAS / RAF产生MAP激酶，2）过表达Zeta-PKC至 上调其信号并研究生化和形态学方面 过表达。 这些研究的长期目标是建立 Zeta-PKC调节NGF依赖和独立的机制 涉及神经元分化的信号传导。 实现这些目标 我们将使用缺陷Zeta-PKC或过表达的PC12单元 这是同工型。 将使用zeta-pkc执行的调节 技术范围：原位活动分析，免疫沉淀 单个信号成分，重构，蛋白质纯化， 肽图和磷酸氨基酸分析。 这些结果 研究将改变我们NGF信号耦合的概念框架和 神经突生长。 而且，这项研究产生的发现将 对阿尔茨海默氏症的研究有直接影响，因为PC12细胞已经 用作阐明淀粉样前体蛋白作用的模型系统 （应用程序）。 PC12细胞中的先前工作已经证明了应用程序能够 增强神经突生长并充当NGF作用的中介。 因此，阐明导致神经突延伸的信号通路 也将直接涉及阿尔茨海默氏症的病因和治疗 因为，PKC和NGF改变的其他人类疾病是 假设发挥作用。