NEURITE PROMOTING EFFECTS OF TPA

TPA 的神经突促进作用

• 批准号: 3402219
• 负责人: KUANG Yu CHEN
• 金额: $ 15.49万
• 依托单位: RUTGERS, THE STATE UNIV OF N.J.
• 依托单位国家: 美国
• 财政年份: 1987
• 资助国家: 美国
• 起止时间: 1987-01-01 至 1989-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3402219
• 关键词: afferent nerve; autoradiography; binding proteins; cell differentiation; cerebellum; cerebral cortex; chick embryo; enzyme induction /repression; enzyme substrate; fluorescent dye /probe; ganglions; gel electrophoresis; glycoproteins; human tissue; laboratory mouse; membrane proteins; neuroblastoma; neurogenesis; neurons; neurotrophic factors; ornithine decarboxylase; phorbols; phosphorylation; plasminogen activator; polyamines; protease inhibitor; protein kinase C; receptor; spinal cord; stainings; tissue /cell culture

Previously, we demonstrated that the phorbol ester, 12-0-tetradecanoyl-phorbol-13-acetate (TPA) was effective in eliciting a dense outgrowth of neurites from 9 day old embryonic sensory ganglia. We will extend our survey of neuronal targets responsive to TPA and attempt to define the molecular mechanisms underlying the TPA effects. We suggest further studies which will compare the neurite outgrowth response of central neuronal tissues including explants from the spinal cord, cerebral cortex and cerebellum. The receptor binding characteristics of sensory neurons from chick embryos ranging from 9-20 days in age will also be studied. Our goal is to extend our previous observations of the neurite promoting effects of TPA so that we can evaluate its action in terms of neuronal target and age specificity. Since the Ca++/phospholipid-dependent protein kinase (protein kinase C, or PK-C) is most likely the cellular phorbol ester receptor within the cell, we will correlate the neurite-promoting effects of TPA with activation of PK-C. We will test if treatment of neuronal cultures with known activators of PK-C such as diacylglycerol and phospholipase C will reproduce the neurite-promoting effects of TPA. Inhibitors of PK-C will also be employed to determine if they are effective in preventing the outgrowth of neurites induced by TPA. To begin formulating the molecular basis of TPA action, we will search for native substrates proteins of PK-C in neuronal cells. Three other TPA effects which may bear relevance to neurite outgrowth will be studied. The induction decarboxylase and plasminogen activator by TPA will be correlated with neurite outgrowth. Changes in the surface glycoproteins of neuronal cells subsequent to TPA treatment will also be examined both morphologically and biochemically. These experiments should provide useful information on the molecular basis of neuritogenesis and the physiological significance of phorbol ester receptors, most likely PK-C, in neuronal development.

先前，我们证明佛波醇酯， 12-0-十四烷酰基-佛波醇-13-乙酸酯（TPA）可有效地诱导 来自9天大的胚胎感觉神经节的神经突的密集生长。 我们 将扩大我们对TPA反应的神经元靶点的调查，并试图 定义TPA效应的分子机制。 我们建议 进一步的研究将比较神经突生长反应， 中枢神经元组织，包括来自脊髓、脑 皮质和小脑。 感觉神经元的受体结合特性 来自9-20天龄的鸡胚的神经元也将被 研究了 我们的目标是扩展我们之前对神经突的观察 促进TPA的效果，以便我们可以评估其作用方面， 神经元靶向和年龄特异性。 由于Ca++/磷脂依赖 蛋白激酶（蛋白激酶C，或PK-C）最有可能是细胞的 佛波酯受体在细胞内，我们将相关的 TPA的促神经突起作用与PK-C的激活。 我们将测试如果 用已知的PK-C活化剂处理神经元培养物， 甘油二酯和磷脂酶C将复制促进神经突的 TPA的影响。 还将使用PK-C抑制剂来确定是否 它们可有效地防止TPA诱导的神经突生长。 为了开始阐明TPA作用的分子基础，我们将寻找 神经元细胞中PK-C的天然底物蛋白。 3其他TPA 将研究可能与神经突生长相关的作用。 的 TPA诱导脱羧酶和纤溶酶原激活剂的作用将相互关联 长出神经突 神经细胞表面糖蛋白的变化 TPA处理后的细胞也将被检查， 形态学和生物化学上。 这些实验应该提供有用的 信息的分子基础的神经突和生理 佛波酯受体，最可能是PK-C，在神经元 发展