We next aimed to further test this by assaying spontaneous action potential (sAP) firing in response to acute 40 mM ethanol application (Supplemental Fig. S2). Action potentials were monitored for 5 min prior to ethanol application to determine the baseline sAP frequency (Supplemental Fig. S2A, B) and amplitude (Supplemental Fig. S2A, C). There were no differences observed between baseline action potential characteristics across genotypes. Acute ethanol was then perfused into the recording chamber and spikes were measured for sAP frequency (Supplemental Fig. S2A, D) and amplitude (Supplemental Fig. S2A & E). Ethanol was equilibrated into the bath for 5 min, and responses were separated into either early (0–2.5 min) or late (2.5–5 min) bins. Following perfusion of 40 mM ethanol, we observed a decrease in the frequency of sAPs for early and late responses, exclusively in N40 AD-iNs, with only a modest decrease observed for D40 AD-iNs (Supplemental Fig. S2D). The late effect of acute ethanol application displayed significantly different genotypic differences between N40 and D40 for sAP frequency (Supplemental Fig. S2D, *p ≤ 0.05). Furthermore, there was no difference
