通过荧光寿命成像使用基于EPAC的FRET传感器记录细胞内cAMP水平。

Recording intracellular cAMP levels with EPAC-based FRET sensors by fluorescence lifetime imaging.

作者信息

Raspe Marcel, Klarenbeek Jeffrey, Jalink Kees

机构信息

Department of Cell Biology and Biochemistry, The Netherlands Cancer Institute, Plesmanlaan 121, 1066 CX, Amsterdam, The Netherlands.

出版信息

Methods Mol Biol. 2015;1294:13-24. doi: 10.1007/978-1-4939-2537-7_2.

DOI:10.1007/978-1-4939-2537-7_2

PMID:25783874

Abstract

Eukaryotic cells use second messengers such as Ca(2+), IP3, cGMP, and cAMP to transduce extracellular signals like hormones, via membrane receptors to downstream cellular effectors. FRET-based sensors are ideal to visualize and measure these rapid changes of second messenger concentrations in time and place. Here, we describe the use of EPAC-based FRET sensors to measure cAMP with spatiotemporal resolution in cells by fluorescence lifetime imaging (FLIM).

摘要

真核细胞利用诸如Ca(2+)、IP3、cGMP和cAMP等第二信使，通过膜受体将激素等细胞外信号转导至下游细胞效应器。基于荧光共振能量转移（FRET）的传感器非常适合在时间和空间上可视化和测量这些第二信使浓度的快速变化。在此，我们描述了基于EPAC的FRET传感器通过荧光寿命成像（FLIM）在细胞中以时空分辨率测量cAMP的方法。