Channel-Based Reporters for cAMP Detection
- 1.5k Downloads
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.
KeywordsCyclic nucleotide-gated channel cAMP GPCR Adenylyl cyclase Phosphodiesterase
This work was supported by NIH P01HL066299, the Center for Lung Biology, and the Colleges of Medicine and Engineering, University of South Alabama.
- 11.Rochais F, Vandecasteele G, Lefebvre F et al (2004) Negative feedback exerted by cAMP-dependent protein kinase and cAMP phosphodiesterase on subsarcolemmal cAMP signals in intact cardiac myocytes: an in vivo study using adenovirus-mediated expression of CNG channels. J Biol Chem 279:52095–52105CrossRefPubMedGoogle Scholar
- 19.Kwan CY, Putney JW (1990) Uptake and intracellular sequestration of divalent cations in resting and methacholine-stimulated mouse lacrimal acinar cells. Dissociation by Sr2+ and Ba2+ of agonist-stimulated divalent cation entry from the refilling of the agonist-sensitive intracellular pool. J Biol Chem 265:678–684PubMedGoogle Scholar
- 28.Molday RS (1998) Photoreceptor membrane proteins, phototransduction, and retinal degenerative diseases: the Friedenwald Lecture. Invest Ophthalmol Vis Sci 39:2493–2513Google Scholar
- 34.Ponimaskin EG, Heine M, Zeug A et al (2007) Monitoring receptor-mediated changes of intracellular camp level by using ion channels and fluorescent proteins as biosensors. In: Chattopadhyay A (ed) Serotonin receptors in neurobiology. CRC Press, Boca Raton, FL, Chapter 2Google Scholar
- 37.Hille B (2001) Ionic channels of excitable membranes. Sinauer Associates, Sunderland, MAGoogle Scholar