Effects of the sulphydryl inhibitor N-ethyl-maleimide on the phrenic nerve and diaphragm muscle of the rat

Abstract

N-Ethyl-maleimide (NEM, 2.5 × 10 −5M) inhibited the compound action potential of the phrenic nerve and increased the spontaneous release of transmitter from the nerve terminals, recorded as miniature endplate potentials. The first effect was the cause of a blockade of the phrenic nerve diaphragm preparation, during indirect stimulation. The left phrenic nerve was more susceptible to inhibition than the right. An increase of the threshold was observed during the progression of the inhibition. The inhibition was not use-dependent and there was no synergistic interaction with the local anaesthetic drug, tetracaine. The inhibition was partly antagonized by di-thio-threitol (3.0 × 10 −3 M). The increase of spontaneous release of transmitter was not accompanied by an increase of the stimulus-evoked release since the amplitude of the endplate potential was not increased and partial inhibition caused by d-tubocurarine or magnesium chloride was not antagonized. When the concentration of NEM was increased to 2.75 × 10 −4 M, the directly-elicited twitches were inhibited, and the baseline tension was increased. This increase of tension was slightly reduced in a preparation depolarized with potassium chloride; a small depolarization could partly explain this effect. It was not reduced by dantrolene or in a calcium-free solution. The inhibition of the twitch and the increased baseline tension (probably a rigor) might be caused by a reduced sensitivity of the contractile proteins for calcium ions and an inhibition of the myosin ATPase activity, respectively.