CILIARY NEUROTROPHIC FACTOR RECEPTORS ON NEURONS

神经元上的睫状神经营养因子受体

• 批准号: 2037498
• 负责人: STANLEY W HALVORSEN
• 金额: $ 18.63万
• 依托单位: STATE UNIVERSITY OF NEW YORK AT BUFFALO
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-09-17 至 2000-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2037498
• 关键词: animal genetic material tag; chickens; developmental genetics; developmental neurobiology; gel mobility shift assay; gene induction /repression; growth factor receptors; neurogenesis; neurogenetics; neurons; neuropharmacology; neurotrophic factors; nucleic acid probes; pharmacogenetics; polymerase chain reaction; receptor binding; receptor expression; tissue /cell culture; transcription factor

DESCRIPTION: (Applicant's Abstract) Cell-cell interactions play important roles in neuronal development, maintenance and regeneration. These actions are believed to be mediated by both electrical activity and neurotrophic factors. The overall goals of this lab are to determine the molecular mechanisms by which these factors can act. For many cholinergic neurons ciliary neurotrophic factor (CNTF) is a candidate molecule participating in these events. CNTF affects the in vitro development of sensory neurons, autonomic neurons and glia, and rescues target-deprived motor and sensory neurons in vivo. The multiple and dramatic responses of chick ciliary ganglion neurons to CNTF coupled with the accessibility and simplicity of this ganglion make it a valuable model system to study CNTF actions in detail, both in vivo and in vitro. During normal development 50% of these neurons will die, but all can be rescued from this fate in cell culture by a select group of agents including CNTF. Receptor(s) and molecular mechanisms responsible for these effects of CNTF have been proposed, but supporting evidence is lacking, nor is it known what the roles of CNTF may be in vivo. Here the applicants outline plans for determining the molecular mechanisms of action of CNTF and related neurokines on ciliary ganglion neurons by taking three different approaches. First, are studies using biochemical and immunological methods to determine the intracellular and molecular pathways activated by CNTF. These studies will use antibodies for STAT transcription factors and nuclear gel shift assays to identify activated transcription elements and their recognition sites on genes. Second, are microinjection studies using STAT blocking antibodies and response element DNA probes injected into CNTF-sensitive neurons to block specific transduction pathways and determine their role in regulating cell survival and neural phenotype. Third are studies aimed at identifying genes regulated by CNTF on ganglion neurons using reverse transcriptase-polymerase chain reaction coupled with differential display to identify novel genes regulated by CNTF. The value of this research is to advance our understanding of the molecular mechanisms and trophic factor signal pathways in the control of neuronal phenotype. The results may also have implications for the role of trophic factors in neural development, neural disease and neural injury as well.

描述：（申请人的摘要）细胞与细胞的相互作用非常重要 在神经元发育、维持和再生中发挥作用。 这些动作 据信是由电活动和神经营养介导的 因素。 该实验室的总体目标是确定分子 这些因素发挥作用的机制。 对于许多胆碱能神经元 睫状神经营养因子（CNTF）是参与参与的候选分子 这些事件。 CNTF影响感觉神经元的体外发育， 自主神经元和神经胶质细胞，并拯救目标剥夺的运动和感觉 体内的神经元。 雏鸡纤毛的多重且戏剧性的反应 神经节神经元到 CNTF 的可访问性和简单性 该神经节使其成为研究 CNTF 行为的有价值的模型系统 体内和体外的细节。 在正常发育期间，其中 50% 神经元会死亡，但在细胞培养中，所有神经元都可以通过一种方法从这种命运中拯救出来： 选择包括 CNTF 在内的代理组。 受体和分子机制 已提出对 CNTF 的这些影响负责，但支持 缺乏证据，也不知道 CNTF 在体内的作用。 申请人在此概述了确定分子机制的计划 CNTF和相关神经因子对睫状神经节神经元的作用 采取三种不同的方法。 首先，是使用生化和 确定细胞内和分子途径的免疫学方法 由 CNTF 激活。 这些研究将使用抗体进行 STAT 转录 因子和核凝胶位移测定来识别激活的转录 元素及其在基因上的识别位点。 二、显微注射 使用 STAT 阻断抗体和反应元件 DNA 探针的研究 注射到 CNTF 敏感神经元中以阻断特定的转导途径 并确定它们在调节细胞存活和神经表型中的作用。 第三是旨在识别神经节中 CNTF 调控基因的研究 神经元使用逆转录酶聚合酶链反应结合 差异显示来识别 CNTF 调控的新基因。 价值 这项研究的目的是增进我们对分子机制的理解 和营养因子信号通路控制神经元表型。 该结果也可能对营养因子在 神经发育、神经疾病和神经损伤。