Simultaneous optical measurements of cytosolic Ca2+ and cAMP in single cells.
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.
15 references, click to expand
-
A new generation of Ca2+ indicators with greatly improved fluorescence properties.J Biol Chem. 1985 Mar 25;260(6):3440-50 PMID: 3838314
-
Novel single chain cAMP sensors for receptor-induced signal propagation.J Biol Chem. 2004 Sep 3;279(36):37215-8 PMID: 15231839
-
Single cell Ca2+/cAMP cross-talk monitored by simultaneous Ca2+/cAMP fluorescence ratio imaging.Proc Natl Acad Sci U S A. 1996 May 14;93(10):4577-82 PMID: 8643446
-
Fluorescent indicators for Ca2+ based on green fluorescent proteins and calmodulin.Nature. 1997 Aug 28;388(6645):882-7 PMID: 9278050
-
Bcl-2-mediated alterations in endoplasmic reticulum Ca2+ analyzed with an improved genetically encoded fluorescent sensor.Proc Natl Acad Sci U S A. 2004 Dec 14;101(50):17404-9 PMID: 15585581
-
Interplay of Ca2+ and cAMP signaling in the insulin-secreting MIN6 beta-cell line.J Biol Chem. 2005 Sep 2;280(35):31294-302 PMID: 15987680
-
Termination of cAMP signals by Ca2+ and G(alpha)i via extracellular Ca2+ sensors: a link to intracellular Ca2+ oscillations.J Cell Biol. 2005 Oct 24;171(2):303-12 PMID: 16247029
-
Fluorescence resonance energy transfer-based assays for the real-time detection of nitric oxide signaling.Methods Enzymol. 2005;396:317-26 PMID: 16291241
-
Biosensors to measure inositol 1,4,5-trisphosphate concentration in living cells with spatiotemporal resolution.J Biol Chem. 2006 Jan 6;281(1):608-16 PMID: 16249182
-
Simultaneous measurements of intracellular cAMP and L-type Ca2+ current in single frog ventricular myocytes.J Physiol. 2001 Jan 1;530(Pt 1):79-91 PMID: 11136860
-
CAMP and Ca2+ interplay: a matter of oscillation patterns.Trends Neurosci. 2003 Feb;26(2):53-5 PMID: 12536124
-
Visualization of the spatial and temporal dynamics of intracellular signaling.Dev Cell. 2003 Mar;4(3):295-305 PMID: 12636912
-
Crosstalk between cAMP and Ca2+ signaling in non-excitable cells.Cell Calcium. 2003 Dec;34(6):431-44 PMID: 14572802
-
FRET imaging.Nat Biotechnol. 2003 Nov;21(11):1387-95 PMID: 14595367
-
Fluorescence ratio imaging of cyclic AMP in single cells.Nature. 1991 Feb 21;349(6311):694-7 PMID: 1847505
Contact
No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong
Qilu Normal University · Genelibs Bioinformatics Lab
750 Shunhua Rd, Jinan
2F, Bldg F, University Science Park
Tel: 0531-88819269
WeChat Official Account
Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.
Business Email
E-mail: [email protected]
Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2
