Contributions of NaV1. 8 and NaV1. 9 to excitability in human induced pluripotent stem-cell derived somatosensory neurons

Abstract

The inhibition of voltage-gated sodium (Na_V) channels in somatosensory neurons presents a promising novel modality for the treatment of pain. However, the precise contribution of these channels to neuronal excitability, the cellular correlate of pain, is unknown; previous studies using genetic knockout models or pharmacologic block of Na_V channels have identified general roles for distinct sodium channel isoforms, but have never quantified their exact contributions to these processes. To address this deficit, we have utilized dynamic clamp electrophysiology to precisely tune in varying levels of Na_V1.8 and Na_V1.9 currents into induced pluripotent stem cell-derived sensory neurons (iPSC-SNs), allowing us to quantify how graded changes in these currents affect different parameters of neuronal excitability and electrogenesis. We quantify and report direct relationships between Na_V1.8 current density and action potential half-width, overshoot, and repetitive firing. We additionally quantify the effect varying Na_V1.9 current densities have on neuronal membrane potential and rheobase. Furthermore, we examined the simultaneous interplay between Na_V1.8 and Na_V1.9 on neuronal excitability. Finally, we show that minor biophysical changes in the gating of Na_V1.8 can render human iPSC-SNs hyperexcitable, in a first-of-its-kind investigation of a gain-of-function Na_V1.8 mutation in a human neuronal background.