CNGA2 contributes to ATP-induced noncapacitative Ca2+ influx in vascular endothelial cells

J Vasc Res. 2010;47(2):148-56. doi: 10.1159/000235969. Epub 2009 Sep 4.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenine / analogs & derivatives
  • Adenine / pharmacology
  • Adenosine Diphosphate / analogs & derivatives
  • Adenosine Diphosphate / pharmacology
  • Adenosine Triphosphate / metabolism*
  • Adenylyl Cyclase Inhibitors
  • Adenylyl Cyclases / metabolism
  • Aminoquinolines / pharmacology
  • Aniline Compounds
  • Animals
  • Calcium Channel Blockers / pharmacology
  • Calcium Signaling* / drug effects
  • Cattle
  • Cells, Cultured
  • Cyclic GMP / analogs & derivatives
  • Cyclic GMP / pharmacology
  • Cyclic Nucleotide-Gated Cation Channels / antagonists & inhibitors
  • Cyclic Nucleotide-Gated Cation Channels / metabolism*
  • Diltiazem / pharmacology
  • Endothelial Cells / drug effects
  • Endothelial Cells / metabolism*
  • Enzyme Inhibitors / pharmacology
  • Imines / pharmacology
  • Male
  • Mice
  • Mice, Inbred C57BL
  • Microscopy, Confocal
  • Purinergic P2 Receptor Antagonists
  • RNA Interference
  • Receptors, Purinergic P2 / metabolism
  • Receptors, Purinergic P2Y1
  • Thionucleotides / pharmacology
  • Time Factors
  • Transfection
  • Vasodilation
  • Xanthenes

Substances

  • 8-(4-chlorophenylthio)guanosine 3',5'-cyclic monophosphorothioate
  • Adenylyl Cyclase Inhibitors
  • Aminoquinolines
  • Aniline Compounds
  • Calcium Channel Blockers
  • Cnga2 protein, mouse
  • Cyclic Nucleotide-Gated Cation Channels
  • Enzyme Inhibitors
  • Fluo 4
  • Imines
  • N(6)-methyl-2'-deoxyadenosine 3',5'-diphosphate
  • P2ry1 protein, mouse
  • Purinergic P2 Receptor Antagonists
  • Receptors, Purinergic P2
  • Receptors, Purinergic P2Y1
  • Thionucleotides
  • Xanthenes
  • 9-(tetrahydro-2-furyl)-adenine
  • Fluo-3
  • Adenosine Diphosphate
  • RMI 12330A
  • Adenosine Triphosphate
  • 6-anilino-5,8-quinolinedione
  • Adenylyl Cyclases
  • Diltiazem
  • Cyclic GMP
  • Adenine