Fig. 3

Effects of prenatal alcohol exposure on circadian expression of epigenetic modulators (transcription activators-MLL1, A–C; P300, D–F; transcription inhibitor-SIRT1, G–I; EZH2, J–L; HDAC3, M–O; JAR1D1, P–R) known to interact with CLOCK:BMAL1 and impose oscillatory patterns to the epigenetic mark at the promoter of PER/CRY gene to maintain circadian rhythmicity. (A,D,G,J,M,P) Showing changes in the rhythmic expression of epigenetic modulators of core clock genes in salivary samples collected at two time points (morning and afternoon) from each AE and control subjects throughout the day and plotted at the time of collections during ZT 3:00 to ZT 11:00. (B,E,H,K,N,Q) Showing changes in the expression of these clock genes when plotted as mean ± SEM at each hour during ZT 3:00 to ZT 11:00 (for n values see Figure legend 1). Data were analyzed using Two-way ANOVA and the interaction between the treatment and time were identified [MLL1: F (8, 83) = 5.711, P = 0.0001; P300: F (8, 83) = 3.316, P = 0.0025; SIRT1: F (8, 83) = 4.739, p = 0.0001; EZH2: F (8, 83) = 4.211, p = 0.0003; HDAC3: F (8, 83) = 4.398, p < 0.0002; JAR1D1: F (8, 83) = 4.525, p = 0.0001]. (C,F,I,L,O,R) Overall Mean ± SEM levels of gene expression in two treatment groups were analyzed using one-way ANOVA. *p < 0.5; ***p < 0.001; ****p < 0.0001. (S) Cosinor analysis was used to determine the rhythmicity (Zero-amplitude test p value < 0.05 identified rhythmicity and the acrophase value shows peak time of the rhythm) of these clock genes. (T) Schematic diagram showing postulated mechanisms by which alcohol exposure (AE) affects epigenetic modifications and alters circadian clock gene (CCG) transcriptions during the circadian cycle in children with FASD. The effects may involve alterations in the Clock/Bmal1 complex-regulated epigenetic modifications by various activators and repressors at the Cry and Per (other clock regulatory genes) promoters to influence their circadian expression. Down arrow indicates inhibition.