Prenatal bisphenol A exposure is associated with sex-specific cerebellar transcriptomic alterations and altered motor performance in rat offspring

Abstract

Autism spectrum disorder (ASD) is a neurodevelopmental condition characterized by a significant male-to-female bias and cerebellar abnormalities. Although bisphenol A (BPA) has been increasingly investigated as a potential environmental risk factor, the molecular mechanisms through which prenatal BPA exposure may affect cerebellar development remain unclear. This study investigated whether prenatal BPA exposure is associated with sex-specific cerebellar transcriptomic alterations and motor-related behavioral outcomes in male and female rat offspring. Eight-week-old female Wistar Furth rats ( Rattus norvegicus ) were assigned to the control or BPA treatment group. After mating, from gestational day 1 (GD1) until parturition, dams in the control group received corn oil vehicle at 20 µL/kg maternal body weight, whereas dams in the BPA treatment group received BPA at 5,000 µg/kg maternal body weight via oral gavage. We performed RNA sequencing (RNA-seq) to profile the offspring cerebellar transcriptome, followed by quantitative RT-PCR (qRT-PCR) assessment of selected candidate genes. Motor behavioral tests were conducted to assess motor coordination, sensorimotor function, locomotor activity, and neuromuscular strength. Bioinformatics analyses were used to evaluate enrichment and overlap with ASD-related gene sets, and molecular docking was used to computationally evaluate potential binding poses of BPA with selected transcription factors. RNA-seq analysis identified BPA-associated gene-expression alterations with sex-specific patterns in male and female offspring. The BPA-responsive gene lists were significantly enriched for SFARI ASD candidate genes, whereas no statistically significant enrichment was observed against the two reanalyzed human ASD cerebellum datasets. Prenatal BPA exposure was associated with alterations in selected motor-test outcomes in a sex-dependent manner, with affected domains differing between male and female offspring. Selected genes, including Dmd, Lrfn5, Ntrk3, Dixdc1 , and Pax6 , showed expression differences alongside selected motor-test alterations. Dmd, Ntrk3, Dixdc1 , and Pax6 were annotated as targets of selected ASD-related transcription factors, and docking analyses generated predicted BPA binding poses for AR, ESR1, RORA, SMAD4, TCF4, YY1, PAX6, and STAT1. Our findings suggest that prenatal BPA exposure is associated with sex-stratified cerebellar molecular alterations and selected motor-related behavioral changes. The candidate gene lists were enriched for ASD-related annotations. Descriptive overlap with genes reported in human ASD cerebellum was observed, but gene-level enrichment against the two reanalyzed human ASD datasets was not statistically significant. These findings nominate transcription factor-related mechanisms for future experimental investigation.