Paradoxical neostigmine-induced TOFfade: on the role of presynaptic cholinergic and adenosine receptors.

Neuromuscular transmission is clinically monitored using the train-of-four ratio (TOFratio), which is the quotient between twitch tension produced by the fourth (T4) and by the first (T1) stimulus within a train-of-four stimulation at 2Hz. Neostigmine causes a paradoxical depression of the TOFratio (TOFfade) that is amplified by intra-arterial atropine in cats. This led us to question the usefulness of the TOFratio as a sole testing element to control neostigmine-induced reversal of neuromuscular transmission block caused by non-depolarizing agents. We hypothesized that the inhibition of cholinesterase activity might increase acetylcholine bioavailability and consequently cholinoceptor activation, leading to concomitant adenosine release from nerve endings and skeletal muscle fibers. The involvement of presynaptic muscarinic and adenosine receptors in neostigmine-induced TOFfade in the rat phrenic nerve diaphragm was investigated. Blockade of adenosine A2A receptors with ZM241385 and of muscarinic M2 receptors with methoctramine or atropine amplified neostigmine-induced TOFfade. Notwithstanding TOFfade amplification, the blockade of M2 or A2A receptors increased both T1 and T4 twitch tensions above control during the first 3min of neostigmine application. Beyond that period, the T1 twitch tension returned to baseline, whereas T4 decreased further until the control value with neostigmine alone. Blockade of M1 receptors by pirenzepine did not change neostigmine-induced TOFfade, unless A2A receptors were concurrently blocked with ZM241385. Data indicate that the paradoxical neostigmine-induced fade involves presynaptic mechanisms that regulate transmitter release and synaptic adenosine accumulation, including the activation of adenosine A2A and muscarinic M2 receptors.

Apamin reduces neuromuscular transmission by activating inhibitory muscarinic M(2) receptors on motor nerve terminals.

This study was undertaken to investigate the mechanism by which the toxin from the bee venom, apamin, might exert beneficial effects in patients suffering from myotonic dystrophy. The effects of apamin were compared with those produced by another potassium channel blocker, 4-aminopyridine, on rat hemidiaphragm preparations stimulated at a 100 Hz frequency via the phrenic nerve. Apamin and 4-aminopyridine increased nerve-evoked tetanic fade without changing the maximal tetanic tension. The inhibitory effect of apamin was mimicked by acetylcholine. In contrast with apamin, 4-aminopyridine increased the amplitude of muscle contractions induced by nerve stimulation at 0.2 Hz frequency. All these compounds were devoid of effect when diaphragm muscle fibres were stimulated directly in the presence of the neuromuscular blocker, D-tubocurarine. The muscarinic M(2) receptor antagonist, methoctramine, prevented the inhibitory effects of both apamin and acetylcholine. Blockade of presynaptic facilitatory muscarinic M(1) and nicotinic receptors respectively with pirenzepine and hexamethonium increased apamin-induced tetanic fade. Data suggest that apamin inhibits neuromuscular transmission by a mechanism independent of the blockade of Ca(2+)-activated K(+) channels, which might involve the activation of inhibitory muscarinic M(2) receptors on motor nerve terminals. Such a mechanism may be the origin of the beneficial effect of apamin controlling muscle excitability in patients suffering from myotonic diseases.