Effects of Some Organic Calcium Antagonists and Other Procedures Affecting Ca2+ Translocation on KCl-induced Contractions in the Rat Vas Deferens

1 Both phasic and tonic responses to KCl 160 mM were reduced by Ca2+ deprivation. After 90 min, the phasic response was abolished but 13 +/- 1.5% of the tonic response remained. This resistant component was still present if the Ca2+-free solution contained EGTA 0.1 mM. The tonic response was more resistant to deprivation in the prostatic half, while the phasic was more resistant in the epididymal half. KCl-induced contractions were completely restored 5 min after readmission of Ca2+. 2 Both the phasic and the tonic responses were reduced on lowering, and increased on raising [Ca2+]0. In 0.1 mM Ca2+, the phasic response was abolished, but 23 +/- 4% of the tonic response remained (mainly attributable to the prostatic half). These resistant contractions indicate that some of the extracellular Ca2+, especially in the prostatic half, is bound with high affinity, probably to the plasma membrane. 3 Incubation with LaCl3 (0.3-10 mM) for 15 min inhibited the phasic response more than the tonic. After incubation for 1 h, 3 mM LaCl3 abolished both phases. It is concluded that La3+ blocks Ca2+ channels most readily when they are opened during the spike. Hydralazine (0.76-5.1 mM) resembled LaCl3 in that it reduced the phasic response with little effect on the tonic. 4 MnCl2 (0.3-10mM) reduced the phasic but increased the tonic response at all concentrations. The augmenting effect may be due to release of intracellular Ca2+ or to inhibition of Ca2+ efflux. 5 The tonic response was inhibited more than the phasic response by nifedipine (0.002-0.01 microM), methoxyverapamil (0.06-2 microM), verapamil (0.2-1 microM), flunarizine (0.2-100 microM) and diazoxide (22-650 microM). With higher concentrations, only flunarizine, remained selective for the tonic response. It is concluded that flunarizine blocks Ca2+ channels most readily when opened during sustained spike-free depolarization. 6 Methoxyverapamil 48 microM and verapamil 100 microM virtually abolished both phases of the contraction to KCl 160 mM, but no more than 80% inhibition could be produced with nifedipine. It is concluded that voltage-sensitive Ca2+ channels exist in two sub-types, one of which is blocked by nifedipine, and both are blocked by verapamil, methoxyverapamil and flunarizine. Nitroprusside 17 microM had no effect on the phasic response but inhibited the nifedipine-resistant component of the tonic response. 7 Increasing [ca2+]0 reversed the effects of verapamil, methoxyverapamil, nifedipine and MnCl2, but not the effects of LaCl3. 8 Dantrolene sodium (1.25-25 microM) had no effect on KCl-induced contractions.