The interaction of diadenosine polyphosphates with P2x-receptors in the guinea-pig isolated vas deferens.
1. The site(s) at which diadenosine 5',5"'-P1,P4-tetraphosphate (AP4A) and diadenosine 5', 5"'-P1,P5-pentaphosphate (AP5A) act to evoke contraction of the guinea-pig isolated vas deferens was studied by use of a series of P2-receptor antagonists and the ecto-ATPase inhibitor 6-N,N-diethyl-D-beta,gamma-dibromomethyleneATP (ARL 67156). 2. Pyridoxalphosphate-6-azophenyl-2',4'-disulphonic acid (PPADS) (300 nM - 30 microM), suramin (3-100 microM) and pyridoxal-5'-phosphate (P-5-P) (3-1000 microM) inhibited contractions evoked by equi-effective concentrations of AP5A (3 microM), AP4A (30 microM) and alpha,beta-methyleneATP (alpha,beta-meATP) (1 microM), in a concentration-dependent manner and abolished them at the highest concentrations used. 3. PPADS was more potent than suramin, which in turn was more potent than P-5-P. PPADS inhibited AP5A, AP4A and alpha,beta-meATP with similar IC50 values. No significant difference was found between IC50 values for suramin against alpha,beta-meATP and AP5A or alpha,beta-meATP and AP4A, but suramin was more than 2.5 times more potent against AP4A than AP5A. P-5-P showed the same pattern of antagonism. 4. Desensitization of the P2xi-receptor by alpha,beta-meATP abolished contractions evoked by AP5A (3 microM) and AP4A (30 microM), but had no effect on those elicited by noradrenaline (100 microM). 5. ARL 67156 (100 microM) reversibly potentiated contractions evoked by AP4A (30 microM) by 61%, but caused a small, significant decrease in the mean response to AP5A (3 microM). 6. It is concluded that AP4A and AP5A act at the P2xi-receptor, or a site similar to the P2xi-receptor, to evoke contraction of the guinea-pig isolated vas deferens. Furthermore, the potency of AP4A, but not AP5A, appears to be inhibited by an ecto-enzyme which is sensitive to ARL 67156.
Westfall
T D
TD
Department of Physiology and Pharmacology, University of Strathclyde, Royal College, Glasgow.
McIntyre
C A
CA
Obeid
S
S
Bowes
J
J
Kennedy
C
C
Sneddon
P
P
D000251:Adenosine Triphosphatases / Q000037:antagonists & inhibitors; D000255:Adenosine Triphosphate / Q000031:analogs & derivatives / Q000494:pharmacology / Q000633:toxicity; D000704:Analysis of Variance; D000818:Animals; D015226:Dinucleoside Phosphates / Q000378:metabolism / Q000494:pharmacology*; D004305:Dose-Response Relationship, Drug; D004791:Enzyme Inhibitors / Q000494:pharmacology; D006168:Guinea Pigs; D007928:Lethal Dose 50; D008297:Male; D009119:Muscle Contraction / Q000187:drug effects; D009638:Norepinephrine / Q000494:pharmacology; D010975:Platelet Aggregation Inhibitors / Q000378:metabolism / Q000494:pharmacology*; D011732:Pyridoxal Phosphate / Q000031:analogs & derivatives / Q000633:toxicity; D018048:Receptors, Purinergic P2 / Q000187:drug effects* / Q000378:metabolism; D015290:Second Messenger Systems; D013498:Suramin / Q000633:toxicity; D014649:Vas Deferens / Q000187:drug effects* / Q000378:metabolism; D014662:Vasoconstrictor Agents / Q000378:metabolism / Q000494:pharmacology*
C092431:6-N,N-diethyl-beta,gamma-dibromomethylene-D-ATP; D015226:Dinucleoside Phosphates; D004791:Enzyme Inhibitors; D010975:Platelet Aggregation Inhibitors; D018048:Receptors, Purinergic P2; D014662:Vasoconstrictor Agents; C077792:pyridoxal phosphate-6-azophenyl-2',4'-disulfonic acid; C012275:P(1),P(5)-di(adenosine-5'-)pentaphosphate; C020733:diadenosine tetraphosphate; D011732:Pyridoxal Phosphate; D013498:Suramin; D000255:Adenosine Triphosphate; D000251:Adenosine Triphosphatases; C002630:alpha,beta-methyleneadenosine 5'-triphosphate; D009638:Norepinephrine