Vasoconstrictor responses to exogenous adenine and pyrimidine nucleotides were measured in endothelium-denuded segments of guinea pig mesenteric vein and compared with responses in mesenteric artery. The rank order of potency for nucleotides in veins was: 2-MeSADP = 2-MeSATP > UTP > ATPgammaS = alpha,betaMeATP > UDP = ATP > ADP >> beta,gamma-D-MeATP = beta,gamma-L-MeATP. In contrast 2-MeSADP, UTP, and UDP were inactive in arteries, and the rank order of potency of other nucleotides differed; that is, alpha,betaMeATP > beta, gamma-D-MeATP > beta,gamma-L-MeATP = ATPgammaS = 2-MeSATP > ATP > ADP. In veins, UTP, ATP, and 2-MeSATP were more efficacious contractile agents than alpha,beta MeATP. In addition, the ability to desensitize responses to these nucleotides and inhibit them with various blockers differed. The response to alpha,betaMeATP in veins exhibited rapid desensitization and was inhibited by pyridoxal-phosphate-6-azophenyl-2',4'-disulfonic acid tetrasodium (PPADS) and suramin. The response to 2-MeSATP in veins did not desensitize; nor was it inhibited by prior alpha,betaMeATP desensitization, but it was inhibited by PPADS, suramin, and the selective P2Y(1) receptor antagonist adenosine 3',5'-bisphosphate (ABP, 10-100 microM). Responses to ATP and UTP in veins did not desensitize and were not inhibited by PPADS, suramin, ABP, or alpha, betaMeATP desensitization. In conclusion, our results suggest that venous contraction to a variety of nucleotides is mediated in large part by P2Y receptors including P2Y(1) receptors and an UTP-preferring P2Y receptor. A small component of contraction also appears to be mediated by P2X(1) receptors. This receptor profile differs markedly from that of mesenteric arteries in which P2X(1) receptors predominate.