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

doi:10.1126/stke.3532006pl6

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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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