Fig. 1

Effects of prenatal alcohol exposure on circadian expression of peripheral circadian markers (A–C) and core clock genes (D–X). (A,D,G,J,M,P,S,T) Showing changes in the rhythmic expression of core clock genes in salivary samples collected at two time points (morning and afternoon) from each AE and control subjects throughout the day (ZT3 to ZT11) and plotted as spline curves (B,E,H,K,N,Q,T,W) 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 in AE (ZT3, n = 3; ZT4, n = 6; ZT5, n = 3; ZT6, n = 4; ZT7, n = 9; ZT8, n = 2; ZT9, n = 10; ZT, n = 3: to ZT11, n = 6) and CONT subjects (ZT3, n = 3; ZT4, n = 7; ZT5, n = 6; ZT6, n = 3; ZT7, n = 5; ZT8, n = 6; ZT9, n = 5; ZT, n = 9: to ZT11, n = 7). Data were analyzed using Two-way ANOVA and the interaction between the treatment and time were identified [BMAL1: F (8, 79) = 3.218, p = 0.0032; CLOCK: F (8, 79) = 2.306, p = 0.0282; PER1: F (8, 79) = 3.446, p = 0.0019; PER2: F (8, 79) = 5.536, p < 0.0001; PER3: F (8, 79) = 3.440, p = 0.0019; CRY1: F (8, 79) = 9.740, p < 0.0001; CRY2: F (8, 79) = 10.62, p < 0.0001; ARR1B: F (8, 83) = 16.60, p < 0.0001]. (C,F,I,L,O,R,U,X) Overall Mean ± SEM levels of gene expression in two groups were analyzed using one-way ANOVA. ***p < 0.001. (Y) 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. (Z) Schematic diagram showing postulated mechanisms by which alcohol exposure (AE) alters circadian oscillation by modifying expression of core clock genes in AE children. The daily circadian oscillation which is maintained by a coordinated transcription-translation feedback loop between the core clock activators (BMAL and CLOCK) and core clock repressors (PER and CRY) is affected by AE. Down arrow indicates inhibition.