1. The role of calcium stores in noradrenaline- (NA) and caffeine-induced contractions of human vas deferens were investigated using ryanodine and cyclopiazonic acid (CPA) in the presence of the calcium antagonist, nifedipine (1 microM) or in calcium-free/EGTA (1 mM) medium. 2. In either media, NA (100 microM) evoked biphasic contractions of longitudinal muscle and tonic circular muscle contractions. Caffeine (20 mM) evoked longitudinal but not circular muscle contractions. 3. Ryanodine (1-30 microM) or CPA (1-30 microM) inhibited contractions of circular muscle, and the initial but not secondary component of longitudinal muscle contraction to NA. 4. In the presence of nifedipine, pre-exposure to caffeine caused a potentiation of circular muscle, and the initial but not secondary longitudinal muscle contractions to NA. The presence of ryanodine or CPA during the caffeine pre-exposures effectively blocked the potentiation of the initial component and reduced the secondary component of the subsequent responses to NA in longitudinal muscle. 5. In calcium-free media, caffeine pre-exposures had little effect on subsequent NA-induced contractions in circular muscle, but reduced both components in longitudinal muscle. The presence of ryanodine or CPA during caffeine pre-exposures produced no further effects on either component of the subsequent NA-induced contraction in longitudinal muscle. 6 In the presence of nifedipine or in calcium-free media, repeated applications of caffeine evoked contractions in longitudinal muscle which were not blocked by either ryanodine or CPA. 7. These results suggest that circular muscle contraction by NA and the initial component of longitudinal muscle to NA both utilize an intracellular pool of calcium that is triggered via a ryanodine-sensitive mechanism and replenished via a CPA-sensitive Ca2+-ATPase. 8. In longitudinal muscle, both the secondary component of its response to NA and contraction to caffeine appear to involve an unusual but pharmacologically distinct (ryanodine- and CPA-insensitive) pathway. 9. The quiescence to caffeine of circular muscle may be caused by a relative absence of the ryanodine- and CPA-insensitive pathway.