CALCIUM INFLUX PATHWAYS IN ALLERGY

描述：(改编自申请人的摘要)持续的钙离子进入 肥大细胞在变态反应的启动和维持中起关键作用 与Fc epsilon RI的配基结合相关的应答。据信， 与Fc epsilon RI结合相关的持续钙内流是 通过开放质膜上的钙通道来调节反应 通过第二信使耗尽一部分钙存储 1，4，5-三磷酸肌醇(IP3)。虽然函数关系 钙储备耗尽和钙进入之间的关系是有充分证据的 对于这种现象，很少或根本没有关于其性质的确凿数据 涉及的相关钙离子通道(称为门店运营通道或 SoC)，或者这些通道作为响应而被门控的分子机制 导致钙库耗尽。此外，尽管最近的数据表明，钙 进入可能受到与第二种生物的产生相关的途径的调节 信使鞘氨醇-1-磷酸，环状ADP-核糖或NAADP， 这些可能的途径与钙储备耗竭的关系，以及 这些途径的共性以及这些特定途径的潜在靶点 第二个信使要么是未知的，要么是有争议的。总而言之，有一个 严重缺乏关于分子机制的具体知识 它调节钙离子进入肥大细胞和其他非兴奋性细胞。 由于钙离子进入对肥大细胞功能的基本重要性， 调查员的实验室已经开始了一系列的实验 鉴定钙离子进入调节蛋白的方法。在预赛中 提供的数据，实验描述了识别和初始 一类新的钙通道家族(CECH蛋白)的特性 它们广泛表达在包括肥大细胞在内的非兴奋性细胞中。在 在当前的应用中，提出了实验来分析它们的功能 肥大细胞和非肥大细胞系中的蛋白质。在具体目标1中，研究 将对野生型切赫的组装和运输进行分析 蛋白质，进行结构/功能分析以确定结构 正确组装和运输所需的功能，以及隔离和 描述切赫相关蛋白在切赫功能中的作用。具体而言 目的2，实验将分析乙酰胆碱在调节钙稳态中的作用 以及通过钙成像和电生理相结合的信号 培养细胞表达特定组合的Cech蛋白的分析 在各种类型的刺激条件下。

DESCRIPTION: (Adapted from the applicant's abstract) Sustained calcium entry in mast cells plays a critical role in the initiation and maintenance of allergic responses associated with ligand binding to Fc epsilon RI. It is believed that the sustained calcium entry associated with engagement of Fc epsilon RI is mediated by the opening of calcium channels in the plasma membrane in response to depletion of a subset of calcium stores by the second messenger inositol-1,4,5-trisphosphate (IP3). Although the functional relationship between calcium store depletion and calcium entry is a well-documented phenomenon, there is little or no definitive data concerning the nature of the relevant calcium channels involved (referred to as Store Operated Channels or SOC), or the molecular mechanisms by which these channels are gated in response to calcium store depletion. Moreover, although recent data suggest that calcium entry may be regulated by pathways associated with the production of the second messengers sphingosine-1-phosphate, cyclic ADP-ribose or NAADP, the relationship between these putative pathways and calcium store depletion, and the generality of these pathways and the potential targets of these specific second messengers are either unknown or controversial. In summary, there is a significant lack of specific knowledge concerning the molecular mechanisms which regulate calcium entry into mast cells and other non-excitable cells. Because of the fundamental importance of calcium entry to mast cell function, the investigator's laboratory has embarked upon a series of experimental approaches to identify calcium entry regulatory proteins. In the preliminary data provided, experiments describe the identification and initial characterization of a novel family of putative calcium channels (CeCH proteins) which are widely expressed in non-excitable cells including mast cells. In the current application, experiments are proposed to analyze the function of these proteins in both mast cell and non-mast cell lines. In specific aim 1, studies will be performed to analyze the assembly and transport of wild type CeCH proteins, to conduct structure/function analyses to identify structural features required for proper assembly and transport, and to isolate and characterize the role of CeCH-associated proteins in CeCH function. In specific aim 2, experiments will analyze CeCH function in regulating calcium homeostasis and signaling through a combination of calcium imaging and electrophysiologic analysis of cultured cells expressing defined combinations of CeCH proteins under various types of stimulus conditions.