Abstract：
Activation of G protein-coupled receptors upon chemoattractant stimulation induces activation of multiple signaling pathways. To fully understand how these signaling pathway coordinates to achieve directional migration of neutrophils, it is essential to determine the dynamics of the spatiotemporal activation profile of signaling components at the level of single living cells. Here, we describe a detailed methodology for monitoring and quantitatively analyzing the spatiotemporal dynamics of 1,4,5-inositol trisphosphate (IP3) in neutrophil-like HL60 cells in response to various chemoattractant fields by applying Förster resonance energy transfer (FRET) fluorescence microscopy.
摘要：
在化学引诱物刺激时G蛋白偶联受体的激活诱导多个信号传导途径的激活。为了充分理解这些信号通路如何协调实现中性粒细胞的定向迁移，在单个活细胞水平上确定信号成分的时空激活谱的动力学是至关重要的。这里,我们描述了通过应用Förster共振能量转移（FRET）荧光显微镜监测和定量分析中性粒细胞样HL60细胞中1,4,5-三磷酸肌醇（IP3）的时空动力学的详细方法。
