Abstract
Background/aims:
ATP can activate several Ca(2+) influx channels in vascular endothelial cells. For example, it stimulates TRPC channels via capacitative and noncapacitative Ca(2+) entry (CCE and non-CCE, respectively) mechanisms; it also directly acts on P2X purinoceptors, resulting in Ca(2+) influx. In the present study, we tested the hypothesis that cyclic nucleotide-gated (CNG) channels also contribute to ATP-induced non-CCE.
Methods:
Two selective inhibitors of CNG channels, L-cis-diltiazem and LY-83583, and CNGA2-specific siRNA were used to study the involvement of CNGA2 in ATP-induced non-CCE in endothelial cells. Ca(2+) influx was studied using Ca(2+)-sensitive fluorescence dyes Fluo-3 and Fluo-4.
Results/conclusion:
L-cis-diltiazem and LY-83583 markedly reduced ATP-induced non-CCE in 3 types of endothelial cells including the H5V endothelial cell line, the primary cultured bovine aortic endothelial cells and the endothelial cells within isolated mouse aortic strips. The CNGA2-specific siRNA also reduced the ATP-induced non-CCE in H5V endothelial cells. The Ca(2+) influx was inhibited by Rp-8-CPT-cAMPS, MDL-12330A, SQ-22536 and MRS-2179, but not by ODQ or NF-157. Taken together, the present study demonstrated that CNGA2 channels contribute to ATP-induced non-CCE in vascular endothelial cells. It is likely that ATP acts through P2Y(1)receptors and adenylyl cyclases to stimulate CNGA2.
Copyright 2009 S. Karger AG, Basel.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Adenine / analogs & derivatives
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Adenine / pharmacology
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Adenosine Diphosphate / analogs & derivatives
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Adenosine Diphosphate / pharmacology
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Adenosine Triphosphate / metabolism*
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Adenylyl Cyclase Inhibitors
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Adenylyl Cyclases / metabolism
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Aminoquinolines / pharmacology
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Aniline Compounds
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Animals
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Calcium Channel Blockers / pharmacology
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Calcium Signaling* / drug effects
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Cattle
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Cells, Cultured
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Cyclic GMP / analogs & derivatives
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Cyclic GMP / pharmacology
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Cyclic Nucleotide-Gated Cation Channels / antagonists & inhibitors
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Cyclic Nucleotide-Gated Cation Channels / metabolism*
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Diltiazem / pharmacology
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Endothelial Cells / drug effects
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Endothelial Cells / metabolism*
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Enzyme Inhibitors / pharmacology
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Imines / pharmacology
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Male
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Mice
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Mice, Inbred C57BL
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Microscopy, Confocal
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Purinergic P2 Receptor Antagonists
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RNA Interference
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Receptors, Purinergic P2 / metabolism
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Receptors, Purinergic P2Y1
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Thionucleotides / pharmacology
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Time Factors
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Transfection
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Vasodilation
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Xanthenes
Substances
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8-(4-chlorophenylthio)guanosine 3',5'-cyclic monophosphorothioate
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Adenylyl Cyclase Inhibitors
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Aminoquinolines
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Aniline Compounds
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Calcium Channel Blockers
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Cnga2 protein, mouse
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Cyclic Nucleotide-Gated Cation Channels
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Enzyme Inhibitors
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Fluo 4
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Imines
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N(6)-methyl-2'-deoxyadenosine 3',5'-diphosphate
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P2ry1 protein, mouse
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Purinergic P2 Receptor Antagonists
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Receptors, Purinergic P2
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Receptors, Purinergic P2Y1
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Thionucleotides
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Xanthenes
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9-(tetrahydro-2-furyl)-adenine
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Fluo-3
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Adenosine Diphosphate
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RMI 12330A
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Adenosine Triphosphate
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6-anilino-5,8-quinolinedione
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Adenylyl Cyclases
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Diltiazem
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Cyclic GMP
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Adenine