Gestational benzyl butyl phthalate exposure impairs sperm capacitation in F1 offspring by altering PKA-associated signaling and tyrosine phosphorylation.

Benzyl butyl phthalate (BBP) is a phthalate plasticizer recognized as an endocrine-disrupting chemical. However, the effects of gestational BBP exposure on sperm function during capacitation in adult offspring remain unclear. This study investigated the effects of gestational BBP exposure on sperm function in adult male offspring, focusing on PKA-associated signaling and protein tyrosine phosphorylation. Pregnant ICR mice were orally administered BBP at 100, 250, or 500 mg/kg/day from gestational day 7 to 20. Adult male F1 offspring were evaluated on postnatal day 78, and sperm function was assessed by measuring motility, kinematic parameters, capacitation status, cell viability, intracellular ATP and reactive oxygen species (ROS) levels, PKA-associated signaling, and protein tyrosine phosphorylation. Gestational BBP exposure significantly reduced body weight, anogenital distance, and testicular size. While cell viability and intracellular ROS levels were unchanged, sperm motility and intracellular ATP levels were significantly reduced, capacitation status was altered, and the acrosome reaction was increased. Furthermore, PKA substrate phosphorylation and protein tyrosine phosphorylation were significantly decreased in spermatozoa. Collectively, these findings demonstrate that gestational BBP exposure impairs sperm function in adult male offspring, potentially through ATP depletion and disruption of capacitation-associated signaling pathways.