ALCOHOL AND TRANSMEMBRANE SIGNALLING

酒精和跨膜信号传导

• 批准号: 3112915
• 负责人: MARIE W WOOTEN
• 金额: $ 10万
• 依托单位: AUBURN UNIVERSITY AT AUBURN
• 依托单位国家: 美国
• 财政年份: 1991
• 资助国家: 美国
• 起止时间: 1991-09-27 至 1994-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3112915
• 关键词: G protein; RNA; acetylcholinesterase; adenylate cyclase; biological signal transduction; cell differentiation; cell growth regulation; cyclic AMP; enzyme induction /repression; ethanol; gene expression; immunocytochemistry; isozymes; membrane permeability; messenger RNA; neurons; neurotrophic factors; phosphorylation; protein kinase C; protein sequence; radiotracer; receptor binding; second messengers; thymidine; tissue /cell culture; tyrosine 3 monooxygenase

PC12 cells have been used as a model system to examine the cellular and molecular mechanisms that govern neuronal differentiation. These cells respond to nerve growth factor (NGF) and acquire the properties of sympathetic neurons. Using PC12 cells, I have observed that ethanol possesses the capability of synergizing with suboptimal concentrations of NGF to induce early neurite extension and subsequent differentiation. Preliminary studies indicate alterations in both protein kinase C and cAMP systems as targets for the effects of ethanol. The long term objective is to develop an understanding of the mechanism by which ethanol interacts with transmembrane signalling pathways to promote gene expression and subsequent differentiation. The specific aims are therefore to define conditions by which ethanol synergizes to promote differentiation by use of pharmacological agents with known specificity and correlate the presence or absence of neurites with indices for differentiation and expression of NGF-specific genes. I propose to detail the contribution of signal transducing pathways which interact to communicate the ethanol transmembrane signal by: a) examining protein kinase C and specifically explore alterations in the isoforms of this kinase brought about by both acute and chronic ethanol exposure; b) analyze activation pathways which either precede PKC activation, ie 'PI-turnover', are a result of PKC activation, i.e. coactivation of other kinase systems such as MAP2 and S6; c) correlate the activation of the adenylate cyclase system as a potential 'cross-talk' regulator between signal transducing pathways; and lastly, d) examine the phosphorylation of tyrosine hydroxylase-as a convergent substrate for multiple kinases.

PC12细胞已被用作模型系统，以检查细胞和 控制神经元分化的分子机制。 这些细胞 响应神经生长因子（NGF）并获得的特性 交感神经元。 使用PC12细胞，我观察到乙醇 具有与次优浓度协同作用的能力 NGF诱导早期神经突扩展和随后的分化。 初步研究表明蛋白激酶C和CAMP的改变 系统作为乙醇影响的目标。 长期目标是 以了解乙醇相互作用的机制 具有跨膜信号通路，以促进基因表达和 随后的分化。 因此，具体的目的是定义乙醇的条件 通过使用药理学剂与 已知特异性并将神经突的存在或不存在与 NGF特异性基因分化和表达的指标。 我 提议详细详细介绍信号传递途径的贡献 互动以传达乙醇跨膜信号：a）检查 蛋白激酶C并专门探索了同工型的改变 这种激酶由急性和慢性乙醇暴露带来。 b） 分析在PKC激活之前的激活途径，即 “ pi turnover”是PKC激活的结果，即其他 激酶系统，例如MAP2和S6； c）关联 腺苷酸环化酶系统作为潜在的“串扰”调节剂 信号传递途径；最后，d）检查 酪氨酸羟化酶 - 作为多个激酶的收敛底物。