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Summary 1 Purinergic transmission from sympathetic nerves in the guinea-pig vas deferens was monitored using intracellular recording techniques. Stimulation of the hypogastric nerve with trains of 15 pulses at 1 Hz evoked excitatory junction potentials (EJPs) which increased in amplitude from the first pulse and reached a maximum after 6–8 pulses. 2 Caffeine (3 and 10 mm), depolarized cells by 5–10 mV and increased the amplitude of the first few EJPs in each train but reduced the maximum amplitude of EJPs late in the train. 3 The adenosine receptor antagonist 8-p-sulphophenyl-theophylline (8-SPT; 30 μm) had no effect on either the resting membrane potential or the EJP amplitude; however, at 100 μmit reduced the amplitude of all EJPs by 5–10%. 4 Adenosine (10 and 30 μm) reduced the amplitude of EJPs in a concentration-dependent manner. The inhibitory effect of adenosine on EJP amplitude was prevented by pretreatment with either caffeine (3 mm) or 8-SPT (30 μm). 5 Ryanodine (30 μm) did not alter EJP amplitude and did not inhibit the enhancement of the first EJP by caffeine (3 mm). Incubation of the tissue with the cell permeable calcium chelator 1-2-bis(o-aminophenoxy)ethane-N,N-N′,N′-tetraacetic acid (BAPT-AM) resulted in a depression of EJP amplitude and a longer time to reach maximum amplitude. In cells that had been exposed to BAPT-AM, caffeine 3 mmstill increased amplitude of EJP early in the train. 6 The phosphodiesterase inhibitor, 3-isobutyl-1-methylxanthine (IBMX; 500 μm), hyperpolarized cells and increased the amplitude of EJP throughout the train of stimulation. In the presence of IBMX, caffeine 3 mmstill depolarized the cells and enhanced the EJP early in the train of stimulation. 7 The findings in this study confirm that caffeine and 8-SPT are effective inhibitors of the actions of adenosine. However, caffeine has an additional action to enhance EJP early during a train of stimulation, which cannot be attributed to blockade of adenosine receptors, but which may be related to inhibition of phosphodiesterase.