Ryanodine Receptor Regulation of Sensory Neuron Function

Ryanodine 受体对感觉神经元功能的调节

基本信息

批准号：
6849129
负责人：
David W Thomas
金额：
$ 22.5万
依托单位：
UNIVERSITY OF THE PACIFIC-STOCKTON
依托单位国家：
美国
项目类别：
财政年份：
2004
资助国家：
美国
起止时间：
2004-09-25 至 2008-08-31
项目状态：
已结题

项目摘要

DESCRIPTION (provided by applicant): The long-term objective of this proposal is to elucidate novel roles played by ryanodine receptors (RyRs) in regulating sensory neuron function. We currently lack a sufficiently detailed understanding of how calcium signaling pathways regulate genetic changes that condition the inflammatory phenotype and influence neuronal survival. Current work suggests that calcium influx channels of the vanilloid receptor (VR) family are important components of inflammatory signaling pathways. Inflammatory signaling mediators such as bradykinin, ATP and nerve growth factor (NGF) regulate VR calcium influx channels, yet it is not known how these receptor systems couple to VR channel activation. Based on recent findings and our preliminary work, our hypothesis is that RyRs are integral regulators of VR or VR-like calcium channels as pivotal couplers linking calcium mobilizing receptors to activation of influx pathways. To begin to dissect the roles of RyRs in neuronal inflammatory responses, we propose to further develop the NG115-401L neuronal cell line as a model cell system for sensory neuron function. Thus, our first specific aim is to characterize the expression profiles of RyRs and calcium channels of the TRP family (including VR channels) in 401L cells and to perform co-immunoprecipitation studies to determine whether RyRs and TRP/TRP-like channels exist as physical complexes. In the second specific aim we take advantage of using the well-characterized homogeneous 401L cell line to produce cell lines deficient in RyR function. RyR deficient 401L cells represent a valuable tool to specifically assess the role of RyRs in calcium signaling pathways elicited by pro-inflammatory agents and the longer-term effects of impaired RyR function on gene expression and growth regulation, which constitutes the third specific aim of the project. A better understanding of RyR function in sensory neurons will greatly augment our ability to understand and rectify flawed RyR activity in inflammatory signaling pathways that lead to conditions of neuropathic pain and inflammation.

描述（由申请人提供）：该提案的长期目标是阐明兰尼碱受体（RyR）在调节感觉神经元功能中所发挥的新作用。目前，我们对钙信号通路如何调节影响炎症表型和影响神经元存活的基因变化缺乏足够详细的了解。目前的工作表明香草酸受体（VR）家族的钙流入通道是炎症信号通路的重要组成部分。缓激肽、ATP 和神经生长因子 (NGF) 等炎症信号传导介质调节 VR 钙流入通道，但尚不清楚这些受体系统如何与 VR 通道激活耦合。根据最近的发现和我们的初步工作，我们的假设是，RyR 是 VR 或 VR 样钙通道的整体调节剂，作为连接钙动员受体与流入途径激活的关键偶联剂。为了开始剖析 RyRs 在神经元炎症反应中的作用，我们建议进一步开发 NG115-401L 神经元细胞系作为感觉神经元功能的模型细胞系统。因此，我们的第一个具体目标是表征 401L 细胞中 RyR 和 TRP 家族钙通道（包括 VR 通道）的表达谱，并进行免疫共沉淀研究以确定 RyR 和 TRP/TRP 样通道是否以物理形式存在。复合物。在第二个具体目标中，我们利用充分表征的同质 401L 细胞系来产生 RyR 功能缺陷的细胞系。 RyR 缺陷的 401L 细胞是一种有价值的工具，可专门评估 RyR 在促炎剂引发的钙信号通路中的作用，以及 RyR 功能受损对基因表达和生长调节的长期影响，这构成了该研究的第三个具体目标项目。更好地了解感觉神经元中的 RyR 功能将极大地增强我们理解和纠正导致神经性疼痛和炎症的炎症信号通路中存在缺陷的 RyR 活性的能力。