Using dynamic clamp to quantify pathological changes in the excitability of primary somatosensory neurons

Abstract

Key points Primary somatosensory neurons normally respond to somatic depolarization with transient spiking but can switch to repetitive spiking under pathological conditions. This switch in spiking pattern reflects a qualitative change in spike initiation dynamics and contributes to the hyperexcitability associated with chronic pain.Neurons can be converted to repetitive spiking by adding a virtual conductance using dynamic clamp. By titrating the conductance to determine how much must be added to cause repetitive spiking, we found that small cells are more susceptible to switching (i.e. required less added conductance) than medium–large cells.By measuring how much less conductance is required to cause repetitive spiking when dynamic clamp was combined with other pathomimetic manipulations (e.g. application of inflammatory mediators), we measured how much each manipulation facilitated repetitive spiking.Our results suggest that many pathological factors facilitate repetitive spiking but that the switch to repetitive spiking requires the cumulative effect of many co‐occurring factors. AbstractPrimary somatosensory neurons become hyperexcitable in many chronic pain conditions. Hyperexcitability can include a switch from transient to repetitive spiking during sustained somatic depolarization. This switch results from diverse pathological processes that impact ion channel expression or function. Because multiple pathological processes co‐occur, isolating how much each contributes to switching the spiking pattern is difficult. Our approach to this challenge involves adding a virtual sodium conductance via dynamic clamp. The magnitude of that conductance was titrated to determine the minimum required to enable rheobasic stimulation to evoke repetitive spiking. The minimum required conductance, termed g¯ Na ∗, was re‐measured before and during manipulations designed to model various pathological processes in vitro. The reduction in g¯ Na ∗ caused by each pathomimetic manipulation reflects how much the modelled process contributes to switching the spiking pattern. We found that elevating extracellular potassium or applying inflammatory mediators reduced g¯ Na ∗ whereas direct hyperpolarization had no effect. Inflammatory mediators reduced g¯ Na ∗ more in medium–large (>30 μm diameter) neurons than in small (⩽30 μm diameter) neurons, but had equivalent effects in cutaneous and muscle afferents. The repetitive spiking induced by dynamic clamp was also found to differ between small and medium–large neurons, thus revealing latent differences in adaptation. Our study demonstrates a novel way to determine to what extent individual pathological factors facilitate repetitive spiking. Our results suggest that most factors facilitate but do not cause repetitive spiking on their own, and, therefore, that a switch to repetitive spiking results from the cumulative effect of many co‐occurring factors.

Ih current stabilizes excitability in rodent DRG neurons and reverses hyperexcitability in a nociceptive neuron model of inherited neuropathic pain

“…The +4 mV liquid junction potential (calculated in Clampex 10) (Vasylyev & Waxman, 2012) between pipette and bath solutions was not compensated. Small DRG neurons (soma diameter 25–30 μm; 27.4 ± 1.4 μm, mean ± SD, n = 24) at 3–7 days in vitro were dynamically clamped (Kemenes et al., 2011; Samu et al., 2012; Sharp et al., 1993a, 1993b; Takkala & Prescott, 2018; Vasylyev et al., 2014) in a whole‐cell configuration using the same pipettes and solutions as for the voltage‐clamp recordings. Ion channel kinetics models were obtained from voltage‐clamp recordings with uncompensated liquid junction potential; however, utilization of these models with dynamic‐clamp was still appropriate since the same liquid junction potential was equally uncompensated both in voltage‐clamp and dynamic‐clamp recordings.…”