K. von Hayn, R. Werthmann, V. Nikolaev, L. Hommers, M. Lohse, and M. Bunemann. Am J Physiol Cell Physiol, 298 (2):
C324-32(February 2010)von Hayn, Kathrin Werthmann, Ruth C Nikolaev, Viacheslav O Hommers,
Leif G Lohse, Martin J Bunemann, Moritz Research Support, Non-U.S.
Gov't United States American journal of physiology. Cell physiology
Am J Physiol Cell Physiol. 2010 Feb;298(2):C324-32. Epub 2009 Nov
4..
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.
von Hayn, Kathrin Werthmann, Ruth C Nikolaev, Viacheslav O Hommers,
Leif G Lohse, Martin J Bunemann, Moritz Research Support, Non-U.S.
Gov't United States American journal of physiology. Cell physiology
Am J Physiol Cell Physiol. 2010 Feb;298(2):C324-32. Epub 2009 Nov
4.
%0 Journal Article
%1 Hayn2010
%A von Hayn, K.
%A Werthmann, R. C.
%A Nikolaev, V. O.
%A Hommers, L. G.
%A Lohse, M. J.
%A Bunemann, M.
%D 2010
%J Am J Physiol Cell Physiol
%K & *Calcium 1/antagonists AMP/*metabolism Adenylate Adrenergic Animals Aorta/enzymology Biosensing Cultured Cyclase/antagonists Cyclic Dose-Response Drug Energy Factors Fluorescence GTP-Binding Gq-G11/*metabolism Humans Inhibitors/pharmacology Interference Isoenzymes/antagonists Isoproterenol/pharmacology Mice Muscle, Muscle/drug Myocytes, Nucleotide Phosphodiesterase Phosphodiesterases, Protein Purinergic/metabolism RNA Relationship, Resonance Signaling/drug Smooth Smooth, Subunits, Techniques Time Transfection Transfer Triphosphate/metabolism Type Uridine Vascular/drug Vasoconstriction Vasodilation Xanthines/pharmacology alpha beta-Agonists/pharmacology effects effects/*enzymology inhibitors/genetics/*metabolism inhibitors/metabolism Receptor Cell
%N 2
%P C324-32
%T Gq-mediated Ca2+ signals inhibit adenylyl cyclases 5/6 in vascular
smooth muscle cells
%U http://www.ncbi.nlm.nih.gov/pubmed/19889965
%V 298
%X 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.
@article{Hayn2010,
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.},
added-at = {2010-12-14T18:12:02.000+0100},
author = {von Hayn, K. and Werthmann, R. C. and Nikolaev, V. O. and Hommers, L. G. and Lohse, M. J. and Bunemann, M.},
biburl = {https://www.bibsonomy.org/bibtex/213f6a36466b98c5c55fe479047213a0f/pharmawuerz},
endnotereftype = {Journal Article},
groups = {private},
interhash = {f026b4ac62780ff20c37dfefb08d5c90},
intrahash = {13f6a36466b98c5c55fe479047213a0f},
issn = {1522-1563 (Electronic) 0363-6143 (Linking)},
journal = {Am J Physiol Cell Physiol},
keywords = {& *Calcium 1/antagonists AMP/*metabolism Adenylate Adrenergic Animals Aorta/enzymology Biosensing Cultured Cyclase/antagonists Cyclic Dose-Response Drug Energy Factors Fluorescence GTP-Binding Gq-G11/*metabolism Humans Inhibitors/pharmacology Interference Isoenzymes/antagonists Isoproterenol/pharmacology Mice Muscle, Muscle/drug Myocytes, Nucleotide Phosphodiesterase Phosphodiesterases, Protein Purinergic/metabolism RNA Relationship, Resonance Signaling/drug Smooth Smooth, Subunits, Techniques Time Transfection Transfer Triphosphate/metabolism Type Uridine Vascular/drug Vasoconstriction Vasodilation Xanthines/pharmacology alpha beta-Agonists/pharmacology effects effects/*enzymology inhibitors/genetics/*metabolism inhibitors/metabolism Receptor Cell},
month = Feb,
note = {von Hayn, Kathrin Werthmann, Ruth C Nikolaev, Viacheslav O Hommers,
Leif G Lohse, Martin J Bunemann, Moritz Research Support, Non-U.S.
Gov't United States American journal of physiology. Cell physiology
Am J Physiol Cell Physiol. 2010 Feb;298(2):C324-32. Epub 2009 Nov
4.},
number = 2,
pages = {C324-32},
shorttitle = {Gq-mediated Ca2+ signals inhibit adenylyl cyclases 5/6 in vascular
smooth muscle cells},
timestamp = {2010-12-14T18:20:49.000+0100},
title = {Gq-mediated Ca2+ signals inhibit adenylyl cyclases 5/6 in vascular
smooth muscle cells},
url = {http://www.ncbi.nlm.nih.gov/pubmed/19889965},
volume = 298,
year = 2010
}