Effect of dithiothreitol on agonist and antagonist actions in frog muscle

The effects of the disulphide bond reducing agent dithiothreitol (DTT) in the frog rectus abdominis preparation have been investigated. DTT, 1 mM, reduced the potency of the monoquaternary agonists acetylcholine, carbachol and tetramethylammonium and the response to electrical field stimulation; the same applied to nicotine, but the action of edrophonium was unaffected and that of the bisquaternary agonist, decamethonium, was increased. The potency of tubocurarine and gallamine as antagonists was unaltered or slightly reduced by DTT when monoquaternary agonists were used and increased when decamethonium was used as agonist. All these effects of DTT were reversed by the oxidizing agent 5-5'dithiobis(2-nitrobenzoic acid) and can be explained by a reduction of a disulphide bond in the vicinity of the anionic site of the nicotinic cholinoceptor. Comparison between these results and published data indicate that there are species differences between nicotinic cholinoceptors and motor endplates. In the guinea-pig ileum preparation DTT reduced the potency of nicotine acting at ganglionic nicotinic cholinoceptors, but had no effect on the agonist response mediated via muscarinic cholinoceptors.