Exposure to chlorpyrifos leads to spindle disorganization and mitochondrial dysfunction of porcine oocytes during in vitro maturation

Abstract

Chlorpyrifos (CPF), as one of the most extensively applied organophosphorus pesticides (OPs) in agricultural and domestic settings, causes a potential threat to human and animal health. Various reproductive toxicities of CPF have been reported, however, little information is available on whether CPF exposure could exert toxic effects on mammalian oocytes. Herein, the effects of CPF on the meiotic maturation and developmental capability of porcine oocytes were investigated, and the possible toxic mechanisms of CPF were also explored. Porcine cumulus-oocyte complexes (COCs) were treated with 0, 5, 10, or 20 μM CPF for 44 h during in vitro maturation (IVM), and the results showed that the first polar body (PB1) extrusion rate was significantly decreased, and the subsequent developmental competence of the resulting metaphase II (MII) oocytes was also impaired when the concentration of CPF reached 10 μM. In addition, a higher percentage of CPF-exposed oocytes were arrested at the anaphase-telophase I (ATI) stage, accompanied by misaligned chromosomes and aberrant spindles. Furthermore, higher levels of ROS and upregulated antioxidant-related genes (CAT, SOD1, SOD2, GPX) were detected in CPF-treated oocytes. Additionally, CPF treatment led to the depolarization of mitochondrial membrane potential (MMP) and the release of cytochrome c (Cyt c). Simultaneously, the apoptotic rate of the oocytes was significantly increased, and the expression levels of Bax and CASPASE3 were significantly upregulated after CFP exposure. Together, exposure to 10 μM CPF can disrupt the meiotic cycle progression, lead to aberrant spindles and mitochondrial dysfunction, which eventually induce oxidative stress and apoptosis in porcine oocytes.