Mechanism of cholinergic inhibition of dog thyroid secretion in vitro.

It has been previously shown that carbamylcholine (10(-5) M) decreases TSH-induced cAMP accumulation and hormone secretion in dog thyroid slices. The mechanism of the latter effect has been investigated in this work. The role of a decrease of cAMP level as the sole mediator of the inhibition of secretion was excluded: the inhibition persisted in the presence of 1-methyl-3-isobutylxanthine at 10(-4) M, which completely abolished the carbamylcholine-induced decrease in cAMP. Moreover, carbamylcholine also inhibited secretion when the slices were incubated with 0.4 mM (Bu)2cAMP. Scanning electron microscopic studies showed that carbamylcholine added at the same time as TSH blocked the formation of pseudopods in response to TSH within 2 min. The kinetic and morphological effects of carbamylcholine added at the same time as, or 90 min after, TSH were similar to those of cytochalasin B (3 micrograms/ml). After carbamylcholine addition at time 90 min, the stimulated secretion rate persisted unchanged for 46 +/- 10 min (mean +/- SD) (n = 6). During this period the colloid droplets disappeared from the cells. Carbamylcholine, like cytochalasin B, did not affect the basal secretion, which is independent of phagocytosis. It is concluded that carbamylcholine (10(-5) M) inhibits stimulated thyroid secretion at a step beyond cAMP accumulation by blocking pseudopod formation and not by inhibiting thyroglobulin hydrolysis or hormone diffusion.