Diadenosine polyphosphates are largely ineffective as agonists at natively expressed P2Y(1) and P2Y(2) receptors on cultured human saphenous vein endothelial cells.
The diadenosine polyphosphates are a group of long-lasting compounds, released into the bloodstream by platelet degranulation. They mediate endothelium-dependent vasodilatation in several animal vascular systems via P2Y receptors coupled to increases in cytoplasmic calcium ([Ca(2+)](c)). However, there is little evidence of diadenosine-mediated vasodilatation in the human vasculature, and a direct interaction with natively expressed P2Y receptors on human endothelium has not been demonstrated. We have therefore studied the effects of diadenosines on primary cultures of human saphenous vein endothelial cells (HSVECs) and related this to the expression of P2Y receptors. HSVECs were loaded with the calcium-sensitive dye fura-2, and nucleotide-stimulated [Ca(2+)](c) responses were recorded. HSVECs responded to 10 microM UTP, ATP and 2-methylthio-ATP but not to UDP. Consistent with the recorded [Ca(2+)](c) responses, RT-PCR analysis of HSVEC RNA amplified specific products for the P2Y(1) and P2Y(2) receptors but not the P2Y(4) and P2Y(6) receptors. HSVECs responded to Ap(3)A with a rise in [Ca(2+)](c), but none of the other diadenosines tested elicited a response. Therefore natively expressed human P2Y(1) and P2Y(2) receptors are insensitive to diadenosine polyphosphates with the exception of Ap(3)A. We would therefore predict that the diadenosine polyphosphates have only a limited vasodilatory role in human saphenous veins.
Conant
A R
AR
Cardiothoracic Centre, Liverpool NHS Trust, UK. conant@liv.ac.uk
Fisher
M J
MJ
McLennan
A G
AG
Simpson
A W
AW
D000255:Adenosine Triphosphate / Q000031:analogs & derivatives* / Q000494:pharmacology*; D020013:Calcium Signaling / Q000187:drug effects*; D002478:Cells, Cultured; D018076:DNA, Complementary / Q000235:genetics; D015226:Dinucleoside Phosphates / Q000494:pharmacology*; D004730:Endothelium, Vascular / Q000166:cytology / Q000187:drug effects*; D006632:Histamine / Q000494:pharmacology; D006801:Humans; D058910:Purinergic P2 Receptor Agonists; D012333:RNA, Messenger / Q000096:biosynthesis; D058472:Receptors, Purinergic P2Y1; D058473:Receptors, Purinergic P2Y2; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012501:Saphenous Vein / Q000166:cytology*; D013329:Structure-Activity Relationship; D013873:Thionucleotides / Q000494:pharmacology*; D014544:Uridine Triphosphate / Q000494:pharmacology*; D014665:Vasodilator Agents / Q000494:pharmacology*
D018076:DNA, Complementary; D015226:Dinucleoside Phosphates; C546382:P2RY1 protein, human; C546385:P2RY2 protein, human; D058910:Purinergic P2 Receptor Agonists; D012333:RNA, Messenger; D058472:Receptors, Purinergic P2Y1; D058473:Receptors, Purinergic P2Y2; D013873:Thionucleotides; D014665:Vasodilator Agents; C012276:diadenosine pyrophosphate; C012275:P(1),P(5)-di(adenosine-5'-)pentaphosphate; C020733:diadenosine tetraphosphate; C011440:diadenosine triphosphate; C070070:diadenosine 5',5''''-P1,P6-hexaphosphate; D006632:Histamine; D000255:Adenosine Triphosphate; D014544:Uridine Triphosphate; C008501:2-methylthio-ATP