Signaling Mechanisms and Mobility of the EGF Receptor

本项目主要研究培养中丝裂原刺激细胞中离子信号的调节。表皮生长因子(EGF)和细胞外ATP都将作为有丝分裂激动剂进行研究。将结合荧光探针成像和膜片钳电生理学对单个细胞离子信号进行研究。我们将考察这些激动剂之间的异源相互作用。将模拟从激活的受体到G蛋白、脂肪酶、激酶和可溶性第二信使的信号转导级联的化学动力学后果。动力学预测将与单细胞数据进行比较，以阐明各种信号成分在初级有丝分裂信号后细胞内钙变化的时间响应模式中的作用。这项研究的重点是表皮生长因子细胞受体激活后的信号转导级联。光学显微镜结合细胞内游离钙离子浓度和膜电位的荧光指示剂将被用来在单个细胞水平上研究这些因素。将构建一个信令级联的数学模型，以帮助理解这一复杂的事件序列。这项研究的结果将有助于理解细胞对外部信号分子反应的分子基础。

This project focuses on the regulation of ionic signaling in mitogen-stimulated cells in culture. Both epidermal growth factor (EGF) and extracellular ATP will be studied as mitogenic agonists. Individual cell ionic signals will be studied by a combination of fluorescent probe imaging and patch clamp electrophysiology. Heterologous interactions between these agonists will be examined. The chemical kinetic consequences of the signal transduction cascade, from activated receptor through G-protein, lipase, kinase, and soluble second messengers, will be modeled. Kinetic predictions will be compared with single cell data to clarify the roles of the various signaling components in the temporal response pattern of intracellular calcium changes subsequent to a primary mitogenic signal. This research focuses on the signal transduction cascade following activation of the cellular receptor for epidermal growth factor. Optical microscopy in combination with fluorescent indicators of cytosolic free calcium ion concentration and of membrane potential will be used to study these factors at the level of a single cell. A mathematical model of the signaling cascade will be constructed to aid in the understanding of this complex sequence of events. The results of this research will contribute to understanding of the molecular basis of the response of cells to external signaling molecules.