单细胞中细胞溶质Ca2+和cAMP的同步光学测量。
Simultaneous optical measurements of cytosolic Ca2+ and cAMP in single cells.
作者信息
Harbeck Mark C, Chepurny Oleg, Nikolaev Viacheslav O, Lohse Martin J, Holz George G, Roe Michael W
机构信息
Department of Medicine, The University of Chicago, Chicago, IL 60637, USA.
出版信息
Sci STKE. 2006 Sep 19;2006(353):pl6. doi: 10.1126/stke.3532006pl6.
Abstract
Understanding the temporal and spatial integration of the Ca2+ and adenosine 3',5'-monophosphate (cAMP) signaling pathways requires concurrent measurements of both second messengers. Here, we describe an optical technique to simultaneously image cAMP and Ca2+ concentration gradients in MIN6 mouse insulinoma cells using Epac1-camps, a Förster (or fluorescence) resonance energy transfer (FRET)-based cAMP biosensor, and Fura-2, a fluorescent indicator of Ca2+. This real-time imaging method allows investigation of the dynamic organization and integration of multiple levels of signal processing in single living cells.
摘要
要理解钙离子（Ca2+）和3',5'-环磷酸腺苷（cAMP）信号通路的时空整合，需要同时测量这两种第二信使。在此，我们描述了一种光学技术，可使用基于Förster（或荧光）共振能量转移（FRET）的cAMP生物传感器Epac1-camps和钙离子荧光指示剂Fura-2，同时对MIN6小鼠胰岛素瘤细胞中的cAMP和Ca2+浓度梯度进行成像。这种实时成像方法能够研究单个活细胞中多级信号处理的动态组织和整合。
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