CILIARY NEUROTROPHIC FACTOR RECEPTORS ON NEURONS

描述：(申请者摘要)细胞间的相互作用很重要 在神经元发育、维持和再生中的作用。这些操作 被认为是由电活动和神经营养共同介导的 各种因素。这个实验室的总体目标是确定分子 这些因素发挥作用的机制。对许多胆碱能神经元来说 睫状神经营养因子(CNTF)是一种候选分子 这些事件。CNTF影响感觉神经元的体外发育， 自主神经细胞和神经胶质细胞，并拯救目标剥夺的运动和感觉 活体中的神经元。雏鸡纤毛的多重戏剧性反应 神经节细胞对CNTF的可及性和简单性 这一神经节使其成为研究CNTF作用的有价值的模型系统 包括体内和体外的详细信息。在正常发育期间，50%的 神经元会死亡，但在细胞培养中，所有人都可以通过一种 选择包括CNTF在内的一组代理。受体(S)及其分子机制 对CNTF的这些影响负责的人已经提出，但支持 目前还缺乏证据，也不知道CNTF在体内可能起到什么作用。 申请者在这里概述了确定分子机制的计划。 CNTF及相关神经因子对睫状神经节神经元作用的研究 采取了三种不同的方法。首先，研究是使用生化和 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 鉴定激活转录的因子和核凝胶移位分析 元素及其在基因上的识别位置。二是微量注射 STAT阻断抗体和反应元件DNA探针的研究 注射入CNTF敏感神经元阻断特定转导通路 并确定它们在调节细胞存活和神经表型中的作用。 第三项研究旨在确定CNTF对神经节的调控基因。 逆转录-聚合酶链式反应偶联检测神经元 差异显示寻找CNTF调控的新基因。它的价值 这项研究的目的是促进我们对分子机制的理解 以及营养因子信号通路在神经元表型调控中的作用。 这一结果也可能对营养因子在 神经发育、神经疾病和神经损伤。

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.