Nerve membrane sodium channels as the target site of brevetoxins at neuromuscular junctions

Actions of two structurally related toxins, T-17 and brevetoxin-B, isolated from the red-tide dinoflagellate, Ptychodiscus brevis, were studied on the giant axon of the squid and the neuromuscular junctions of the frog and rat. T-17 toxin caused a large increase in the frequency of miniature endplate potentials at nanomolar concentrations. In one typical case with a frog endplate, the frequency increased from 1.9 s-1 before application of 3.5 nM T-17 to 69.3 s-1 within 5 min after application. In the rat muscle, the mean frequency increased from 1.39 s-1 in control to 11.93 s-1 after application of 23.2 nM T-17. The increase in miniature endplate potential frequency was reversed by the addition of 1 microM tetrodotoxin, and was not observed in a solution containing elevated Mg2+ and reduced Ca2+ concentrations. External or internal application of T-17 toxin (2-5 microM) or brevetoxin-B (10-30 microM) to intact or internally perfused squid axons caused a depolarization of the membrane. This depolarization was abolished by the removal of external Na+ or by addition of tetrodotoxin to the external solution. In voltage clamped squid giant axons, exposure to T-17 toxin or brevetoxin-B increased the non-inactivating component of the tetrodotoxin-sensitive sodium current. The sodium current was activated at potentials 15 to 40 mV more negative than control. It is proposed that these toxins modify a fraction of the sodium channels to a form which opens at potentials more negative than normal and which inactivates to a lesser extent. This mechanism would predict a depolarization of the nerve membrane at the neuromuscular junction, thus explaining the increased discharge of transmitter.