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 Forster
(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.
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