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Ryanodine受体（RYRS）在调节感觉神经元功能中扮演的新作用。我们目前缺乏对钙信号通路如何调节遗传变化的足够详细的理解，从而调节炎症表型并影响神经元存活。当前的工作表明，香草素受体（VR）家族的钙涌入通道是炎症信号通路的重要组成部分。炎症信号介质（例如心动激肽，ATP和神经生长因子（NGF））调节VR钙涌入通道，但尚不清楚这些受体系统如何与VR通道激活。根据最新发现和我们的初步工作，我们的假设是Ryrs是VR或VR样钙通道的整体调节剂，作为关键耦合器，将动员钙的受体与涌入途径的激活联系起来。为了开始剖析Ryrs在神经元炎症反应中的作用，我们建议进一步发展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的是表征401L细胞中TRP家族（包括VR通道）的RYRS和钙通道的表达谱，并进行共免疫沉淀研究，以确定RYRS和TRP/TRP样通道是否作为物理复合物存在。在第二个特定目的中，我们利用使用良好的均质401L细胞系来产生缺陷RYR功能的细胞系。 RYR缺乏401L细胞代表了一种有价值的工具，可以专门评估RYR在钙信号传导途径中的作用，该钙信号传导途径是由促炎剂引起的，以及RYR受损功能对基因表达和生长调节的长期影响，这构成了该项目的第三个特定目标。对感官神经元中的RYR功能的更好理解将大大增强我们理解和纠正炎症信号传导途径中RYR活性的能力，从而导致神经性疼痛和炎症状况。