Ion Channel-Based Reporters for cAMP Detection

Methods Mol Biol. 2022:2483:265-279. doi: 10.1007/978-1-0716-2245-2_17.

Abstract

In the last 20 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 using cyclic nucleotide (CNG) channel-based probes. These probes take advantage of the ability of CNG channels to transduce small changes in cAMP concentration into ionic flux through channel pores that can be readily detected by measuring Ca2+ and/or Mn2+ influx or by measuring ionic currents.

Keywords: Adenylyl cyclase; Cyclic nucleotide-gated channel; GPCR; Phosphodiesterase; cAMP.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Calcium / metabolism
  • Cell Membrane / metabolism
  • Cyclic AMP* / metabolism
  • Cyclic Nucleotide-Gated Cation Channels* / metabolism
  • Signal Transduction

Substances

  • Cyclic Nucleotide-Gated Cation Channels
  • Cyclic AMP
  • Calcium