Using cell surface biotinylation and Western blotting, we investigated the extent to which native P2X₁ receptors in rat vas deferens are internalised after exposure to agonist. Exposure to 100 μM α,β‐meATP 30 min prior and during a 10 min biotinylation period resulted in a ∼50% reduction in the amount of biotinylated P2X₁ receptor indicating that activation of the receptor by agonist induces receptor internalisation. Furthermore, biotinylation under saturating conditions suggests that once internalised, a rapid recycling of P2X₁ receptor back to the cell surface occurs. The physiological implications of these mechanisms in terms of receptor function are discussed.