Prenatal bisphenol A exposure causes sperm quality and functional defects via Leydig cell impairment and meiosis arrest in mice offspring.

Bisphenol A (BPA), widely used in plastic production, acts as an environmental endocrine disruptor which is harmful to male reproductive health. However, the specific mechanisms through which prenatal BPA exposure disrupts spermatogenesis in offspring, particularly in terms of Leydig cell dysfunction and meiotic progression, remain poorly understood. To address this gap, we constructed a mouse model with BPA lowest Observed Adverse Effect Level (LOAEL: 50 mg/kg bw/day) exposure from embryonic day (ED) 0.5 to 18.5. Our results demonstrated that prenatal BPA exposure significantly decreased serum testosterone levels, testis weight, sperm count, motility parameters, and acrosomal integrity. Furthermore, it arrested the meiotic transition from zygotene to pachytene spermatocytes, leading to reduced sperm fertility characterized by reduced sperm-egg binding capacity and abnormal early embryonic cleavage in the male offspring. Importantly, prenatal BPA exposure significantly reduced the expression of PCNA (a marker of germ cell proliferation), SYCP3 (a meiosis regulator), and Vimentin (a blood-testis barrier component), collectively indicating impaired spermatogenesis in offspring testes. Additionally, prenatal BPA exposure dramatically reduced Leydig cell numbers and increased apoptosis, marked by BAX/BCL2 up-regulation, which mechanistically explains the observed testosterone reduction. In vitro experiments corroborated these effects: BPA exposure concentration-dependently inhibited Leydig cell proliferation, induced G0/G1 phase arrest, and downregulated testosterone synthesis molecules (Hsd3b1, Hsd17b3, Star, Cyp11a1, Cyp17a1). Quantitative proteomics identified 234 differentially expressed proteins (97 downregulated, 137 upregulated) in BPA-exposed Leydig cells. Bioinformatics analysis revealed that down-regulated proteins were mainly related to steroid hormone receptor activity, estrogen response element binding, and centrosome duplication processes, while the up-regulated proteins were mainly involved in oxygen binding and ROS metabolic process. Conclusively, prenatal BPA exposure impaired offspring male fertility via multi-faceted mechanisms: sperm quality defects, steroidogenic disruption, and meiotic arrest. This study advances the understanding of BPA transgenerational reproductive toxicity and underscores the need to mitigate prenatal exposure risks.