To examine whether chronic alcohol treatment produced changes in NMDA subunit mRNA expression, 15–18 week old human iPS-derived neural cultures derived from 4 alcoholic and 3 non-alcoholic donor subjects were fed daily with media containing 0 or 50mM alcohol and the levels of expression relative to sham treated cells were examined for the NMDA receptor subunit genes GRIN1 (NR1 subunit), GRIN2A, (NR2A subunit), GRIN2B, (NR2B subunit) and GRIN2D (NR2D subunit) (Figure 4). For neural cultures derived from alcoholic subjects, significant treatment effects were seen after 7 days of alcohol exposure for GRIN1 (F(1,37)=10.8, p=0.002), GRIN2A (F(1,39)=13.5, p=0.001), and GRIN2D (F(1,37)=16.1, p<0.001) mRNA levels. A trend towards significance was seen for GRIN2B (F(1,37)=3.7, p=0.06). Following a 24-hour withdrawal period, the expression of GRIN1 in alcoholic-derived cultures remained significantly elevated (F(1,40)=5.0, p=0.03), the expression of GRIN2A continued to increase (F(1,40)=33.6, p<0.001), and the expression of GRIN2B and GRIN2D returned to baseline levels (F(1,43)=0.2, p=0.64, and F(1,40)=0.6, p=0.44, respectively). In contrast, following 7-day alcohol exposure of neural cultures derived from non-alcohol dependent subjects there was no change in the expression of the four
