Fig. 1

TCS downregulates RA signaling, disrupts synaptic plasticity and induces autistic behaviors in offspring rats. a Chemical structure of TCS. b Concurrent treatment with TCS (12.50 µM) for 12 h abolished the increase of RARE-hrGFP reporter expression (n = 22, 25, 22). The expression of hrGFP is driven by three copies of RARE upstream of TK promoter. Neuronal activity blockade by APV (100 µM) and TTX (1 µM) treatment for 24 h, or acute RA (10 µM) treatment for 8 h, significantly increased the expression of RARE-hrGFP (n = 25, 27, 23), but had no effect on the control reporter that did not have RARE (Supplementary information, Fig. S2). The mean values of normalized GFP expression were 100.00, 189.89, 157.81, 79.54, 90.77, 88.39, respectively. c, d Treatment of APV and TTX or RA (1 µM) significantly induced synaptic upscaling. Addition of TCS disrupted the synaptic scaling, manifested as reduced mEPSC amplitudes (c, n = 13, 12, 13, 8; mean values of amplitude were 13.40, 11.30, 18.12, 14.89, respectively; d, n = 9, 17, 16, 18; mean values were 11.42, 12.12, 14.12, 11.67, respectively). Representative mEPSC traces of PFC neurons were shown in Supplementary information, Fig. S4. e A scheme for TCS administration. Starting from E7.5, wild-type pregnant Sprague-Dawley rats were orally administered with TCS (50 mg/kg/d) until weaning at PND21. Offspring rats were subjected to behavioral tests at the age of 7–8 weeks. f–h Pups from mother rats exposed to TCS manifested autistic-like behaviors. They tended to spend more time with familiar subject in social novelty tests (f), more time with object in sociability tests (g), and longer time on self-grooming (h), compared with pups in control group (CON). CON group, n = 38; 50 mg/kg/d TCS group, n = 39. i Representative western blots of RARα and RARβ proteins using lyzed PFC tissues from two offspring rats of the control or TCS group. Data are means ± SEM. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001, n.s., not significant. One-way ANOVA with Bonferroni post hoc test was used to analyze fluorescence assay and electrophysiology results. Self-grooming time and preference index were analyzed by unpaired two-tailed Student’s t-test with Welch’s correction. For three-chamber experiments, two-way repeated-measures analysis of variance (ANOVA) was used. Detailed statistical analysis results were presented in Supplementary information, Tables S1 and S2