Activation of adenylyl cyclase causes stimulation of adenosine receptors

  • Background/Aims: Signaling of Gs protein-coupled receptors (GsPCRs) is accomplished by stimulation of adenylyl cyclase, causing an increase of the intracellular cAMP concentration, activation of the intracellular cAMP effectors protein kinase A (PKA) and Epac, and an efflux of cAMP, the function of which is still unclear. Methods: Activation of adenylyl cyclase by GsPCR agonists or cholera toxin was monitored by measurement of the intracellular cAMP concentration by ELISA, anti-phospho-PKA substrate motif phosphorylation by immunoblotting, and an Epac-FRET assay in the presence and absence of adenosine receptor antagonists or ecto-nucleotide phosphodiesterase/pyrophosphatase2 (eNPP2) inhibitors. The production of AMP from cAMP by recombinant eNPP2 was measured by HPLC. Extracellular adenosine was determined by LC-MS/MS, extracellular ATP by luciferase and LC-MS/MS. The expression of eNPP isoenzymes 1-3 was examined by RT-PCR. The expression of multidrug resistance protein 4 was suppressed by siRNA. Results: Here we show that the activation of GsPCRs and the GsPCRs-independent activation of Gs proteins and adenylyl cyclase by cholera toxin induce stimulation of cell surface adenosine receptors (A2A or A2B adenosine receptors). In PC12 cells stimulation of adenylyl cyclase by GsPCR or cholera toxin caused activation of A2A adenosine receptors by an autocrine signaling pathway involving cAMP efflux through multidrug resistance protein 4 and hydrolysis of released cAMP to AMP by eNPP2. In contrast, in PC3 cells cholera toxin- and GsPCR-induced stimulation of adenylyl cyclase resulted in the activation of A2B adenosine receptors. Conclusion: Our findings show that stimulation of adenylyl cyclase causes a remarkable activation of cell surface adenosine receptors.

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Author:Thomas Pleli, Antonia Mondorf, Nerea Ferreiros, Dominique Jeanette ThomasORCiDGND, Karel Dvorak, Ricardo M. Biondi, Dagmar Meyer zu HeringdorfORCiDGND, Stefan ZeuzemORCiDGND, Gerd GeisslingerORCiDGND, Herbert Zimmermann, Oliver WaidmannORCiDGND, Albrecht Piiper
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Parent Title (English):Cellular physiology and biochemistry
Place of publication:Basel
Document Type:Article
Year of Completion:2018
Year of first Publication:2018
Publishing Institution:Universitätsbibliothek Johann Christian Senckenberg
Release Date:2018/05/22
Tag:Adenylyl cyclase; MRP4; PKA; adenosine receptors; cAMP; eNPP2
Page Number:13
First Page:2516
Last Page:2528
This article is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License (CC BY-NC-ND) ( Usage and distribution for commercial purposes as well as any distribution of modified material requires written permission.
Institutes:Biowissenschaften / Biowissenschaften
Medizin / Medizin
Wissenschaftliche Zentren und koordinierte Programme / Interdisziplinäres Zentrum für Neurowissenschaften Frankfurt (IZNF)
Dewey Decimal Classification:6 Technik, Medizin, angewandte Wissenschaften / 61 Medizin und Gesundheit / 610 Medizin und Gesundheit
Licence (German):License LogoCreative Commons - Namensnennung-Nicht kommerziell - Keine Bearbeitung 4.0