Benzo[a]pyrene disrupts LH/hCG-dependent mouse Leydig cell steroidogenesis through receptor/Gαs protein targeting

• Published in: Scientific reports 2024-01, Vol.14 (1), p.844-844, Article 844
• Main Authors: Lazzaretti, Clara, Roy, Neena, Paradiso, Elia, Capponi, Chiara, Ferrari, Tommaso, Reggianini, Francesca, Sperduti, Samantha, Perri, Carmela, Baschieri, Lara, Mascolo, Elisa, Varani, Manuela, Canu, Giulia, Trenti, Tommaso, Nicoli, Alessia, Morini, Daria, Iannotti, Francesca, Villani, Maria Teresa, Vicini, Elena, Simoni, Manuela, Casarini, Livio
• Format: Article
• Language: English
• Subjects: 631/45/776/1174; 692/163/2743/2730; Animal models; Animals; Benzo(a)pyrene; Benzo(a)pyrene - toxicity; Biological Assay; Bioluminescence; Blotting, Western; Cell culture; Chorionic Gonadotropin - pharmacology; Cyclic AMP response element-binding protein; Endocrine disruptors; Energy transfer; Female; Gonadotropins; Humanities and Social Sciences; Humans; Immunoassays; Leydig Cells; Luteinizing hormone; Male; Mice; multidisciplinary; Phosphorylation; Pituitary (anterior); Polycyclic aromatic hydrocarbons; Progesterone; Proteins; Pyrene; Science; Science (multidisciplinary); Steroidogenesis; Steroidogenic acute regulatory protein; Toxicity; Western blotting
• ISSN: 2045-2322; 2045-2322
• DOI: 10.1038/s41598-024-51516-7

Steroidogenesis of gonadal cells is tightly regulated by gonadotropins. However, certain polycyclic aromatic hydrocarbons, including Benzo[a]pyrene (BaP), induce reproductive toxicity. Several existing studies have considered higher than environmentally relevant concentrations of BaP on male and female steroidogenesis following long-term exposure. Also, the impact of short-term exposure to BaP on gonadotropin-stimulated cells is understudied. Therefore, we evaluated the effect of 1 nM and 1 µM BaP on luteinizing hormone/choriogonadotropin (LH/hCG)-mediated signalling in two steroidogenic cell models, i.e. the mouse tumor Leydig cell line mLTC1, and the human primary granulosa lutein cells (hGLC) post 8- and 24-h exposure. Cell signalling studies were performed by homogeneous time-resolved fluorescence (HTRF) assay, bioluminescence energy transfer (BRET) and Western blotting, while immunostainings and immunoassays were used for intracellular protein expression and steroidogenesis analyses, respectively. BaP decreased cAMP production in gonadotropin-stimulated mLTC1 interfering with Gαs activation. Therefore, decrease in gonadotropin-mediated CREB phosphorylation in mLTC1 treated with 1 μM BaP was observed, while StAR protein levels in gonadotropin-stimulated mLTC1 cells were unaffected by BaP. Further, BaP decreased LH- and hCG-mediated progesterone production in mLTC1. Contrastingly, BaP failed to mediate any change in cAMP, genes and proteins of steroidogenic machinery and steroidogenesis of gonadotropin-treated hGLC. Our results indicate that short-term exposure to BaP significantly impairs steroidogenic signalling in mLTC1 interfering with Gαs. These findings could have a significant impact on our understanding of the mechanism of reproductive toxicity by endocrine disruptors.