FIGURE 3.

Methadone exposure suppresses the development of the passive neuronal properties. (A) Bright field images of organoids in suspension in control and methadone treatment groups from 12-week till 16-week. 4 weeks treatment of methadone at 1 and 10 μM do not significantly alter organoid diameters. (B) Schematic of the time course for the culture and methadone treatment of cortical organoids in the dish. Whole-cell patch-clamp recordings were performed for every 4-week from 12-week, till 24-week. (C–E) Scatter-plots of the C_m, input resistance (R_in) and resting membrane potential (V_rest) recording of neurons from cortical organoids during the early development (12- to 24-week) in the absence and presence of 1 or 10 μM methadone. In each scatter plot, Black, Blue, and Red dots denote untreated, 1 and 10 μM methadone group, respectively. Graphs display mean and SEM. n = 10–29 neurons (7–17 organoids). Two-way ANOVA followed by Tukey’s multiple comparisons test: (C) F_{(6, 162)} = 6.26, p < 0.0001; (D) F_{(6, 162)} = 1.92, p = 0.08; (E) F_{(6, 162)} = 6.11, p < 0.0001. Control groups at 16-week, 20-week, 24-week were compared to 12-week group, ^##p < 0.01, ^####p < 0.0001. Each methadone-treated groups were compared to the control group at the same age, *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.