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| Content Provider | World Health Organization (WHO)-Global Index Medicus |
|---|---|
| Author | Von Hayn, Kathrin Werthmann, Ruth C. Nikolaev, Viacheslav O. Hommers, Leif G. Lohse, Martin J. Bünemann, Moritz |
| Description | Country affiliation: Germany Author Affiliation: von Hayn K ( University of Marburg, Institute of Pharmacology and Toxicology, Karl-von-Frisch-Strasse 1, 35033 Marburg, Germany.) |
| Abstract | cAMP and Ca(2+) are antagonistic intracellular messengers for the regulation of vascular smooth muscle tone; rising levels of Ca(2+) lead to vasoconstriction, whereas an increase of cAMP induces vasodilatation. Here we investigated whether Ca(2+) interferes with cAMP signaling by regulation of phophodiesterases (PDEs) or adenylyl cyclases (ACs). We studied regulation of cAMP concentrations by Ca(2+) signals evoked by endogenous purinergic receptors in vascular smooth muscle cells (VSMCs). The fluorescence resonance energy transfer (FRET)-based cAMP sensor Epac1-camps allowed the measurement of cAMP levels in single-living VSMCs with subsecond temporal resolution. Moreover, in vitro calibration of Epac1-camps enabled us to estimate the absolute cytosolic cAMP concentrations. Stimulation of purinergic receptors decreased cAMP levels in the presence of the beta-adrenergic agonist isoproterenol. Simultaneous imaging of cAMP with Epac1-camps and of Ca(2+) with Fura 2 revealed a rise of intracellular Ca(2+) in response to purinergic stimulation followed by a decline of cAMP. Chelation of intracellular Ca(2+) and overexpression of Ca(2+)-independent AC4 antagonized this decline of cAMP, whereas pharmacological inhibition of Ca(2+)-activated PDE1 had no effect. AC assays with VSMC membranes revealed a significant attenuation of isoproterenol-stimulated cAMP production by the presence of 2 muM Ca(2+). Furthermore, small interfering RNA (siRNA) knockdown of AC5 and AC6 (the two ACs known to be inhibited by Ca(2+)), significantly reduced the decrease of cAMP upon purinergic stimulation of isoproterenol-prestimulated VSMCs. Taken together, these results implicate a Ca(2+)-mediated inhibition of AC5 and 6 as an important mechanism of purinergic receptor-induced decline of cAMP and show a direct cross talk of these signaling pathways in VSMCs. |
| File Format | HTM / HTML |
| ISSN | 03636143 |
| e-ISSN | 15221563 |
| Journal | American Journal of Physiology - Cell Physiology |
| Issue Number | 2 |
| Volume Number | 298 |
| Language | English |
| Publisher | American Physiological Society |
| Publisher Date | 2010-02-01 |
| Publisher Place | United States |
| Access Restriction | Open |
| Subject Keyword | Discipline Cell Biology Adenylate Cyclase Metabolism Calcium Signaling Cyclic Amp Gtp-binding Protein Alpha Subunits, Gq-g11 Isoenzymes Muscle, Smooth, Vascular Enzymology Myocytes, Smooth Muscle Adenylyl Cyclase Inhibitors Genetics Adrenergic Beta-agonists Pharmacology Animals Aorta Biosensing Techniques Drug Effects Cells, Cultured Cyclic Nucleotide Phosphodiesterases, Type 1 Antagonists & Inhibitors Dose-response Relationship, Drug Fluorescence Resonance Energy Transfer Isoproterenol Mice Phosphodiesterase Inhibitors Rna Interference Receptors, Purinergic Time Factors Transfection Uridine Triphosphate Vasoconstriction Vasodilation Xanthines Research Support, Non-u.s. Gov't |
| Content Type | Text |
| Resource Type | Article |
| Subject | Cell Biology Physiology |
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