Pharmacological and histochemical evidence for P2X receptors in human umbilical vessels

Abstract

The presence of P2X purinoceptors in human umbilical vessels were studied with organ bath recording, radioligand binding assays, autoradiography, and immunohistochemistry. In isolated umbilical arteries and veins from normal term pregnancy, both ATP and α,β-methylene ATP caused concentration-dependent contractions. ATP-induced responses were blocked by desensitisation with α,β-methylene ATP. However, both the ATP- and α,β-methylene ATP-induced responses were not antagonised by suramin. No significant difference in responses was observed in the vessels with or without endothelial cells. Radioligand binding assays using [ 3 H ]α,β-methylene ATP showed the presence of a population of high-affinity binding sites in both the arteries and veins. The K d values of the binding sites were 2.77±1.10 nM for the arteries, and 3.23±1.22 nM for the veins. The maximum binding site densities were 634±237 and 947±308 fmol/mg protein for the arteries and the veins, respectively. Autoradiographic localisation with [ 3 H ]α,β-methylene ATP demonstrated that the specific binding sites were only distributed over the smooth muscle cells of the vessels. Immunohistochemical studies with specific polyclonal antibodies against P2X 1–6 receptors showed that positive immunostaining was also restricted to smooth muscle cells. Antibodies against P2X 1 receptors produced the strongest signals, while antibodies against the other five P2X subtypes produced much weaker signals. The results in the present study indicate the existence of P2X purinoceptors in the smooth muscle of human umbilical vessels. Their physiological functions remain to be studied.