Conway Bruce R, Demarest Keith T
Department of Drug Discovery, R. W. Johnson Pharmaceutical Research Institute, Raritan, New Jersey, USA.
Recept Channels. 2002;8(5-6):331-41.
Plasma membrane-associated G protein-coupled receptors (GPCRs) initiate the transmission of multiple intracellular signals leading to a myriad of physiological and pathophysiological effects. The downstream signaling events associated with occupation of the GPCR and activation of the G-protein include the generation of numerous second messenger molecules to provide the necessary signal amplification within the appropriate intracellular compartment to transmit a specific signal from the cell surface to the cell interior. The complex process of signal transmission also requires a series of highly orchestrated events which includes the translocation of cellular proteins to discreet intracellular destinations. A better understanding of these events has made it possible to design assays to examine multiple endpoints within whole cells. In this review we describe recent advances in assay biology and instrumentation useful for broadening our understanding of the molecular events associated with GPCR activation. This review will focus on novel cell-based approaches using fluorescent biosensors, such as fluorescent dyes and fluorescent protein tags, to generate information-rich data from multiple cellular targets--a process that has been referred to as high-content screening. High-content screening applications will be discussed as they pertain to specific signal transduction cascades initiated upon GPCR activation and examples of specific biosensors will be provided.
质膜相关的G蛋白偶联受体（GPCRs）启动多种细胞内信号的传递，导致无数的生理和病理生理效应。与GPCR的占据和G蛋白的激活相关的下游信号事件包括产生众多第二信使分子，以便在适当的细胞内区室中提供必要的信号放大，从而将特定信号从细胞表面传递到细胞内部。信号传递的复杂过程还需要一系列高度协调的事件，这包括细胞蛋白向离散的细胞内目的地的转运。对这些事件的更好理解使得设计检测全细胞内多个终点的分析方法成为可能。在本综述中，我们描述了分析生物学和仪器方面的最新进展，这些进展有助于拓宽我们对与GPCR激活相关的分子事件的理解。本综述将聚焦于使用荧光生物传感器（如荧光染料和荧光蛋白标签）的新型基于细胞的方法，以从多个细胞靶点生成信息丰富的数据——这一过程被称为高内涵筛选。将讨论高内涵筛选应用与GPCR激活后引发的特定信号转导级联的相关性，并提供特定生物传感器的实例。
