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Channel-Based Reporters for cAMP Detection

  • Thomas C. RichEmail author
  • Wenkuan Xin
  • Silas J. Leavesley
  • Mark S. Taylor
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1294)

Abstract

In the last 15 years, tremendous progress has been made in the development of single-cell cAMP sensors. Sensors are based upon cAMP-binding proteins that have been modified to transduce cAMP concentrations into electrical or fluorescent readouts that can be readily detected using patch clamp amplifiers, photomultiplier tubes, or cameras. Here we describe two complementary approaches for the detection and measurement of cAMP signals near the plasma membrane of cells. These probes take advantage of the ability of cyclic nucleotide-gated (CNG) channels to transduce small changes in cAMP concentrations into ionic flux through channel pores that can be readily detected by measuring Ca2+ and/or Mn2+ influx or by measuring ionic currents.

Keywords

Cyclic nucleotide-gated channel cAMP GPCR Adenylyl cyclase Phosphodiesterase 

Notes

Acknowledgments

This work was supported by NIH P01HL066299, the Center for Lung Biology, and the Colleges of Medicine and Engineering, University of South Alabama.

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Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  • Thomas C. Rich
    • 1
    • 2
    Email author
  • Wenkuan Xin
    • 3
  • Silas J. Leavesley
    • 1
    • 2
    • 4
  • Mark S. Taylor
    • 5
  1. 1.Department of PharmacologyUniversity of South AlabamaMobileUSA
  2. 2.Center for Lung BiologyUniversity of South AlabamaMobileUSA
  3. 3.Department of Drug Discovery and Biomedical SciencesSouth Carolina College of PharmacyMobileUSA
  4. 4.Department of Chemical and Biomolecular EngineeringUniversity of South AlabamaMobileUSA
  5. 5.Department of PhysiologyUniversity of South AlabamaMobileUSA

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