The effects of some agonists acting at P2 purinoceptors on guinea-pig isolated ileum longitudinal smooth muscle have been examined. The preparation contracted in response to ATP, alpha,beta-methylene ATP and 2-methylthio ATP, but not UTP. In this respect, alpha,beta-methylene ATP and 2-methylthio ATP were approximately equipotent and both were 10-50 times more active than ATP. Responses to alpha,beta-methylene ATP, but not 2-methylthio ATP or ATP, were antagonised by atropine and tetrodotoxin, suggesting that alpha,beta-methylene ATP activates cholinergic nerves in the ileum, whilst the other two compounds act on the smooth muscle. Two other purine nucleotide analogues, beta,gamma-methylene ATP and beta,gamma-imido ATP, did not cause contraction. However, both compounds antagonised responses to alpha,beta-methylene ATP, but not those to 2-methylthio ATP. Suramin antagonised responses to both alpha,beta-methylene ATP and 2-methylthio ATP, whilst Cibacron blue was without effect on responses to either agonist. We conclude that the purinoceptor on cholinergic nerves has some of the characteristics of the P2x purinoceptor, whilst the purinoceptor on ileal smooth muscle has some of the characteristics of the P2Y purinoceptor. However, further work will be necessary before definitive classification is possible.