Fig. 4.

Cortical neurons from offspring of BPA-fed mouse dams show Kcc2 promoter repression. (A) Sexually dimorphic delay in chloride shift of primary cortical neurons at DIV4 derived from P0 mouse pups, born to BPA or corn oil-exposed dams. (Right) Quantitation of mRNA levels: Note increased difference for female neurons. (B) Significant methylation increase relative to vehicle in the CpG dinucleotides with equal net effect for both sexes. At least 25 clones were sequenced per CpG dinucleotide. P values determined by Fisher’s exact test. (C) Significant decrease of Kcc2 expression in mixed primary cortical neurons. Decitabine (2 µM) rescues the effect of BPA; DIV4, n = 3 independent experiments, *P < 0.01 (D, Top) Design of 2.5-kb mouse Kcc2–LUC construct (33). (Middle) Significant decrease of Kcc2 promoter-Luc activity in slice cortical culture in response to BPA. Decitabine (2 µM) rescues the effect of BPA, DIV3; n = 4 independent experiments, *P < 0.05. (Bottom) Representative luminescence heatmap of DIV3 organotypic cortical slice culture prepared from offspring of BPA-fed dams. Note increased luminescence in response to decitabine treatment.