Proceedings of the National Academy of Sciences of the United States of America •
Voltage-gated sodium channel Nav1.8 plays a crucial role in regulating excitability of small dorsal root ganglion (DRG) neurons and is an emerging target for pain therapeutics. Using dynamic clamp, we systematically manipulated Nav1.8 conductance to assess its impact on action potential (AP) electrogenesis, rheobase, and repetitive firing in native rat DRG neurons and those expressing the gain-of-function Nav1.7L858H mutation which underlies inherited erythromelalgia, a human genetic pain disorder. Our findings reveal that the Nav1.8 contribution to net sodium current is highly correlated with AP voltage threshold. Nav1.8 conductance regulated AP overshoot and voltage threshold without significantly affecting undershoot or resting membrane potential. We identified two populations of wild-type DRG neurons: strong responders (50% of cells), which exhibited substantial rheobase modulation with alterations in Nav1.8 conductance, and weak responders (50% of cells), which remained largely unaffected. In hyperexcitable Nav1.7L858H-expressing neurons, partial Nav1.8 subtraction (50%) restored rheobase above control levels in 63% of cells. However, weak responders (37%) remained hyperexcitable. The effect of Nav1.8 subtraction in responsive neurons supports the conclusion that Nav1.8 inhibition can reduce neuropathic pain. However, the presence of weakly responsive DRG neurons suggests that other channels might need to be targeted for full pain relief.
Harty TP, Dib-Hajj SD, Tyrrell L, Blackman R, Hisama FM , et al.
The Journal of neuroscience : the official journal of the Society for Neuroscience •
Inherited erythromelalgia/erythermalgia (IEM) is a neuropathy characterized by pain and redness of the extremities that is triggered by warmth. IEM has been associated with missense mutations of the voltage-gated sodium channel Na(V)1.7, which is preferentially expressed in most nociceptive dorsal root ganglia (DRGs) and sympathetic ganglion neurons. Several mutations occur in cytoplasmic linkers of Na(V)1.7, with only two mutations in segment 4 (S4) and S6 of domain I. We report here a simplex case with an alanine 863 substitution by proline (A863P) in S5 of domain II of Na(V)1.7. The functional effect of A863P was investigated by voltage-clamp analysis in human embryonic kidney 293 cells and by current-clamp analysis to determine the effects of A863P on firing properties of small DRG neurons. Activation of mutant channels was shifted by -8 mV, whereas steady-state fast inactivation was shifted by +10 mV, compared with wild-type (WT) channels. There was a marked decrease in the rate of deactivation of mutant channels, and currents elicited by slow ramp depolarizations were 12 times larger than for WT. These results suggested that A863P could render DRG neurons hyperexcitable. We tested this hypothesis by studying properties of rat DRG neurons transfected with either A863P or WT channels. A863P depolarized resting potential of DRG neurons by +6 mV compared with WT channels, reduced the threshold for triggering single action potentials to 63% of that for WT channels, and increased firing frequency of neurons when stimulated with suprathreshold stimuli. Thus, A863P mutant channels produce hyperexcitability in DRG neurons, which contributes to the pathophysiology of IEM.
