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Sci. STKE, 19 September 2006
Vol. 2006, Issue 353, p. pl6
[DOI: 10.1126/stke.3532006pl6]

PROTOCOLS

Simultaneous Optical Measurements of Cytosolic Ca2+ and cAMP in Single Cells

Mark C. Harbeck1, Oleg Chepurny2, Viacheslav O. Nikolaev3, Martin J. Lohse3, George G. Holz2, and Michael W. Roe1*

1Department of Medicine, The University of Chicago, Chicago, IL 60637, USA.
2Department of Physiology and Neuroscience, New York University School of Medicine, New York, NY 10016, USA.
3Institute of Pharmacology and Toxicology, University of Würzburg, D-97078 Würzburg, Germany.

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.

*Corresponding author. Department of Medicine, MC-1027, The University of Chicago, 5841 South Maryland Avenue, Chicago, IL 60637, USA. Telephone, 773-702-4965; fax, 773-834-0486; e-mail, mroe{at}medicine.bsd.uchicago.edu

Citation: M. C. Harbeck, O. Chepurny, V. O. Nikolaev, M. J. Lohse, G. G. Holz, M. W. Roe, Simultaneous Optical Measurements of Cytosolic Ca2+ and cAMP in Single Cells. Sci. STKE 2006, pl6 (2006).

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