Navß subunits modulate the inhibition of Nav1.8 by the analgesic gating modifier µO-conotoxin MrVIB.

ABSTRACT

Voltage-gated sodium channels (VGSCs) consist of a pore-forming α-subunit and regulatory ß-subunits. Several families of neuroactive peptides of Conus snails target VGSCs, including µO-conotoxins and µ-conotoxins. Unlike µ-conotoxins and the guanidinium alkaloid saxitoxin (STX), which are pore blockers, µO-conotoxins MrVIA and MrVIB inhibit VGSCs by modifying channel gating. µO-MrVIA/B can block Na(v)1.8 (a tetrodotoxin-resistant isoform of VGSCs) and have analgesic properties. The effect of Na(v)ß-subunit coexpression on susceptibility to block by µO-MrVIA/B and STX has, until now, not been reported. Here, we show that ß1-, ß2-, ß3-, and ß4-subunits, when individually coexpressed with Na(v)1.8 in Xenopus laevis oocytes, increased the k(on) of the block produced by µO-MrVIB (by 3-, 32-, 2-, and 7-fold, respectively) and modestly decreased the apparent k(off). Strong depolarizing prepulses markedly accelerated MrVIB washout with rates dependent on ß-subunit coexpression. Thus, coexpression of ß-subunits with Na(v)1.8 can strongly influence the affinity of the conopeptide for the channel. This observation is of particular interest because ß-subunit expression can be dynamic, e.g., ß2-expression is up-regulated after nerve injury (J Neurosci, 2510970-10980, 2005); therefore, the effectiveness of a µO-conotoxin as a channel blocker could be enhanced by the conditions that may call for its use therapeutically. In contrast to MrVIB's action, the STX-induced block of Na(v)1.8 was only marginally, if at all, affected by coexpression of any of the ß-subunits. Our results raise the possibility that µO-conotoxins and perhaps other gating modifiers may provide a means to functionally assess the ß-subunit composition of VGSC complexes in neurons.