Electropysiological analysis of the neurotoxic action of a funnel-webspider toxin, delta-Atracotoxin-Hv1a, on insect voltage-gated Na+ channels

摘要

The effects of delta -ACTX-Hv1a, purified from the venom of the funnel-web spider Hadronyche versuta, were studied on the isolated giant axon and dorsal unpaired median (DUM) neurones of the cockroach Periplaneta americana under current- and voltage-clamp conditions using the double oil-gap technique for single axons and the patch-clamp technique for neurones, In parallel, the effects of the toxin were investigated on the excitability of rat dorsal root ganglion (DRG) neurones. In both DRG and DUM neurones, delta -ACTX-Hv1a induced spontaneous repetitive firing accompanied by plateau potentials. However, in the case of DUM neurones, pIateau action potentials were facilitated when the membrane was artificially hyperpolarized, In cockroach giant axons, delta -ACTX-Hv1a also produced plateau action potentials, but only when the membrane was pre-treated with 3-4 diaminopyridine, Under voltage-clamp conditions, delta -ACTX-Hv1a specifically affected voltage-gated Naf channels in both axons and DUM neurones. Both the current/voltage and conductance/ voltage curves of the delta -ACTX-Hv1a-modified inward current were shifted 10 mV to the left of control curves. In the presence of delta -ACTX-Hv1a, steady-state Na+ channel inactivation became incomplete, causing the appearance of a non-inactivating component at potentials more positive than -40mV, The amplitude of this non-inactivating component was dependent on the holding potential, From this study, it is concluded that, in insect neurones, delta -ACTX-Hv1a mainly affects Na+ channel inactivation by a mechanism that differs slightly from that of scorpion alpha -toxins.