Regulation of P2y-purinoceptor-mediated prostacyclin release from human endothelial cells by cytoplasmic calcium concentration.
1. ATP and ATP analogues induced prostacyclin (PGI2) secretion from human cultured umbilical vein endothelial cells. 2. The threshold active concentration for ATP was less than or equal to 1 microM. The rank order of potency of analogues was 2-chloroadenosine 5'-triphosphate (2-ClATP) greater than 2-methylthioadenosine 5'-triphosphate (2-MeSATP) greater than ATP greater than ADP, while adenosine 5'-(alpha,beta-methylene)triphosphonate, AMP and adenosine were inactive, indicating the presence of P2y-purinoceptors. 3. In contrast to their actions on P2y-receptors in guinea-pig taenia coli, isopolar analogues of 2-methylthioadenosine 5'-(beta, gamma-methylene)triphosphonate were less effective than ATP. 4. ATP and ATP analogues increased intracellular free calcium ions, [Ca2+]i, giving a rapid transient peak due predominantly to release from intracellular stores, followed by a maintained steady-state elevated level due to influx. 5. The dose-response curves for peak [Ca2+]i induced by ATP, 2-ClATP and 2-MeSATP were very similar to those for PGI2 production. 6. Elevations of [Ca2+]i, above a threshold value of 0.8-1 microM, were necessary for PGI2 production in response to P2y-receptor activation. 7. The dose relationships between PGI2 release and peak [Ca2+]i were equivalent whether [Ca2+]i was raised by ionomycin or via P2y-receptor activation by ATP or 2-ClATP, indicating that elevations of [Ca2+]i provide the major, if not the exclusive intracellular pathway for P2y-purinoceptor-mediated PGI2 synthesis.
Carter
T D
TD
Section of Vascular Biology, MRC Clinical Research Centre, Harrow, Middx.
Hallam
T J
TJ
Cusack
N J
NJ
Pearson
J D
JD
D000255:Adenosine Triphosphate / Q000031:analogs & derivatives / Q000494:pharmacology; D002118:Calcium / Q000378:metabolism*; D002478:Cells, Cultured; D003593:Cytoplasm / Q000378:metabolism; D004730:Endothelium, Vascular / Q000187:drug effects / Q000502:physiology*; D011464:Epoprostenol / Q000378:metabolism*; D006801:Humans; D011983:Receptors, Purinergic / Q000187:drug effects / Q000502:physiology*
D011983:Receptors, Purinergic; D000255:Adenosine Triphosphate; D011464:Epoprostenol; D002118:Calcium