Molecular insights into the effects of tetrachlorobisphenol A on puberty initiation in Wistar rats

Tetrachlorobisphenol A (TCBPA) is the chlorinated derivative of bisphenol A (BPA). Several studies have found that BPA adversely affects the reproductive activity largely through binding to estrogen receptors and the critical period of BPA exposure advances the vaginal opening time in the female offspring via the kisspeptin/G protein-coupled receptor 54 (KGG) system. However, whether TCBPA can affect puberty initiation via KGG and the roles of estrogen receptors in this process remain unknown. Therefore, this study investigated the influence of TCBPA on the onset time of puberty in Wistar rats and the related molecular mechanisms by combing in vitro GT1–7 cells and molecular docking. In female Wistar rats, TCBPA at ≥100 mg/kg bw/day (49.2 μmol/L in rat body) markedly advanced vaginal opening time and increased serum levels of follicle-stimulating hormone (FSH), luteinizing hormone (LH), and gonadotropin-releasing hormone (GnRH). It also increased the relative gene expression of LH receptor (LHR), GnRH1, and FSH receptor (FSHR) in hypothalamic-pituitary-gonadal (HPG) axis tissues. In GT1–7 cells, TCBPA increased genes and proteins associated with KGG pathway and activated the extracellular-regulated protein kinase 1/2 (Erk1/2) and phosphatidylinositol-3-kinase/protein kinase B (PI3K/Akt) pathways via G protein-coupled estrogen membrane receptor 1 (GPER1) and estrogen receptor alpha (ERα). Docking analyses supported its interactions with GPER1 and ERα, and treatment with specific inhibitors of ERα- and GPER1-modulated PI3K/Akt and Erk1/2 signaling suppressed its effects. Taken together, TCBPA-induced advancement of puberty initiation in Wistar rats thus results primarily from increased LH, GnRH, and FSH secretion together with GnRH1, FSHR, and LHR upregulation driven by ERα- and GPER1-modulated Erk1/2 and PI3K/Akt signaling. Our results provide new molecular insights into the reproductive toxicity of EDCs.