Evidence that 2‐methylthioATP and 2‐methylthioADP are both agonists at the rat hepatocyte P2Y 1 receptor

In the absence of selective antagonists, pharmacological characterization of P2Y receptor subtypes has relied heavily upon their distinct agonist profiles. 2‐methylthioADP (2‐MeSADP) is a selective agonist for the P2Y 1 receptor. The agonist action of 2‐MeSATP at the P2Y 1 receptor has recently been questioned. The effects of both 2‐MeSADP and 2‐MeSATP have been studied on rat hepatocytes injected with the bioluminescent Ca 2+ indicator, aequorin. Single hepatocytes generate series of repetitive transients in cytosolic free calcium concentration ([Ca 2+ ] i ) when stimulated with agonists acting through the phosphoinositide signalling pathway. The transients induced by 2‐MeSADP and 2‐MeSATP in the same cell were indistinguishable, indicating that they act at a common receptor. In contrast the transients evoked by ATP and UTP had very different profiles. Treatment of 2‐MeSATP with an ATP‐regenerating system to remove contaminating 2‐MeSADP did not abolish its agonist activity. Application of the P2Y 1 antagonist, adenosine‐3′‐phosphate‐5′‐phosphate (A3P5P) inhibited the transients induced by both 2‐MeSADP and 2‐MeSATP. In contrast the transients induced by ATP and UTP were enhanced by the addition of A3P5P. These results indicate that both 2‐MeSADP and 2‐MeSATP are agonists at the rat hepatocyte P2Y 1 receptor.British Journal of Pharmacology (2000) 130 , 664–668; doi: 10.1038/sj.bjp.0703350