The inhibition by three modulators (oxotremorine, noradrenaline, morphine) of acetylcholine release from the myenteric plexus preincubated with [3H]choline was investigated at different stimulation frequencies and calcium concentrations. Moreover, [3H]acetylcholine release evoked by a low (0.1 Hz) or a high (10 Hz) stimulation rate was investigated at different calcium concentrations either in the absence or presence of scopolamine. A reduced calcium concentration (0.6 mmol/l) inhibited acetylcholine release more at 0.1 Hz (74% ± 3%) than at 10 Hz (44% ± 8%). Scopolamine enhanced the stimulated acetylcholine release at a calcium concentration of 1.8 mmol/l. At calcium concentrations higher than 1.8 mmol/l scopolamine failed to enhance transmitter release markedly. A reduction of the calcium concentration (< 1.8 mmol/l) significantly enhanced the effect of scopolamine, when acetylcholine release was evoked at 0.1 Hz. Oxotremorine (10 μmol/l) completely suppressed acetylcholine release at 1 Hz (120 pulses). When 120 pulses were applied at 10 Hz the maximal effect was only a 64% inhibition and the concentration-response curve was significantly shifted to the right. However, after a reduction of both the train length or the calcium concentration oxotremorine produced a complete inhibition of acetylcholine release evoked at 10 Hz. In contrast to the effect of oxotremorine, the concentration-response curves for morphine and noradrenaline were similar at 1 Hz and 10 Hz. Following conclusions can be drawn: 1. The present findings fit into the concept that residual calcium accumulates in the nerve terminal during 10 Hz stimulation. 2. The results obtained with scopolamine and oxotremorine are consistent with the view that muscarine autoreceptor activation triggers a reduction of the intraneuronal availability of calcium for the stimulus-secretion coupling. 3. The presynaptic effect of morphine and partly that of noradrenaline might be mediated by a different mechanism, probably by a reduction of release sites.

The myenteric plexus-longitudinal muscle preparation of the guinea-pig ileum was preincubated with [3H]choline, then superfused and stimulated electrically (1 Hz 120 pulses). Oxotremorine reduced the evoked outflow of [3H]acetylcholine in a concentration-dependent manner. Each of the six antagonists (scopolamine, methylatropine, trihexyphenidyl, 4-DAMP, clozapine, pirenzipine) produced parallel shifts of the concentration-response curves for the prejunctional effects of oxotremorine. Similarly, in contraction experiments, the antagonists competitively antagonized the postjunctional responses to oxotremorine. The pre- and postjunctional pA2 values did not differ significantly for any of the antagonists. It is concluded that pre- and postjunctional muscarinic receptors in the guinea-pig ileum are pharmacologically similar.

The effects of pirenzepine and scopolamine on presynaptic muscarine receptors (mediating inhibition of acetylcholine release) and postsynaptic receptors (mediating smooth muscle contraction) were compared in the myenteric plexus-longitudinal muscle preparation of the guinea pig. The preparation was preincubated with 3H-choline and subsequently superfused with Tyrode's solution. Field stimulation (1 Hz; 2 min) caused an outflow of 3H-acetylcholine that was depressed by the muscarinic agonist oxotremorine. Both pirenzepine and scopolamine produced shifts to the right of the concentration-response curve for the inhibitory effect of oxotremorine. Similarly, in contraction experiments, pirenzepine and scopolamine competitively antagonized the responses to oxotremorine. From the dose ratios pA2 values for pre- and postsynaptic effects were calculated. The presynaptic pA2 values for pirenzepine (6.90) and scopolamine (9.07) did not differ significantly from the respective postsynaptic pA2 values (6.66 and 9.40). Thus, pirenzepine like scopolamine does not distinguish between muscarine receptors that mediate contraction and those which mediate inhibition of acetyl-choline release.