Electrophysiological analysis of the inactivation of sympathetic transmitter in the guinea-pig vas deferens

1. The properties of junction potentials evoked by nerve stimulation and by local application of drugs, and currents evoked by nerve stimulation, in the smooth muscle cells of the guinea-pig vas deferens have been investigated. The effects of temperature on these responses have been studied using intracellular and extracellular recording. 2. Local, brief (5-15 ms) application of 10(-4) M-adenosine-5'-triphosphate (ATP) from glass micropipettes onto the surface of the vas deferens, using pressure pulses (103-206 kPa), elicited a depolarization of the smooth muscle cell membranes which closely resembled the nerve stimulation-evoked excitatory junction potential (EJP). 3. Local application of 10(-4) M-noradrenaline (NA) failed to produce any detectable membrane potential response. Junction potentials elicited by a mixture of 10(-4) M-ATP and 10(-4) M-NA (ratio by volume 1:50) in the drug ejection micropipette were similar in shape to those evoked by ATP alone. 4. Cooling the tissue from 35 to 25 degrees C did not significantly alter resting membrane potentials but resulted in a significant prolongation of the rising and decaying phases of the EJPs. Fifty per cent decay times for EJPs at 35 and 25 degrees C were (mean +/- S.D.) 236 +/- 20 and 434 +/- 30 ms respectively (P less than 0.01). 5. Extracellularly recorded excitatory junction currents (EJCs) elicited by nerve stimulation, believed to reflect the transmembrane current underlying the EJPs, were prolonged in parallel at low temperatures (50% decay times of EJCs at 35 and 25 degrees C: 11.73 +/- 3.94 and 26.15 +/- 8.4 ms, respectively, P less than 0.01). 6. Junction potentials evoked by locally applied, exogenous ATP were also significantly prolonged by cooling (50% decay times: 663 +/- 88 ms at 35 degrees C and 1955 +/- 79 ms at 25 degrees C, P less than 0.01). 7. Bath application of 10(-6) M-alpha,beta-methylene ATP, the enzymatically stable, desensitizing analogue of ATP, reversibly abolished nerve-evoked EJPs. Local application of 10(-6) M-alpha,beta-methylene ATP led to a prolonged depolarization of the smooth muscle cells lasting between 20 and 60 s. 8. Junction potentials elicited by locally applied alpha,beta-methylene ATP were not prolonged or otherwise significantly altered on cooling. The durations of the depolarizations were 46.0 +/- 12.1 s at 35 degrees C and 43.4 +/- 10.6 s at 25 degrees C (P greater than 0.1).