Figure 6.
Functional characterization of hPSC-derived nociceptors (WA09)
(A) Calcium flux analysis after stimulation of nociceptors with KCL, ATP, capsaicin, mustard oil, and menthol (n = 8, mean ± SD). Data shown are from eight different wells of one representative experiment.
(B) Automated MEA showing that nociceptors are activated by specific ligands.
(C and D) MEA experiment showing that nociceptors are stimulated by a temperature increase from 37 to 40°C and are presensitized after treatment with oxaliplatin or PGE2 (n = 3, mean ± SD). Data shown are from three different wells of one representative experiment.
(E) Typical action potential in nociceptors evoked by current injection. Arrow indicates the threshold voltage (−35 mV).
(F) Collected results for passive and active electrical properties of the neurons. Open circles show measurements from individual cells and closed circles show mean ± SEM (n = 53 for cell capacitance and resting potential; n = 25 for action potential peak and width).
(G) Enhancement of excitability by the Kv1-inhibitor α-dendrotoxin (DTX).
(H) Automated single-cell patch-clamp (Sophion Qube) recording showing effect of 10 nM Protoxin-II and 1 μM TTX (in the continuing presence of Protoxin-II) on sodium current in an hPSC-derived nociceptor.
(I) Collected results for percentage inhibition of sodium current by Protoxin-II alone and by Protoxin-II plus TTX (217 neurons analyzed).