Perfluorooctane Sulfonate (PFOS) and Perfluorohexane Sulfonate (PFHxS) Alters Protein Phosphorylation, Increase ROS Levels and DNA Fragmentation during In Vitro Capacitation of Boar Spermatozoa

SIMPLE SUMMARY: Perfluorinated compounds are synthetic chemicals, with a wide variety of applications like firefighting foams, food packaging, additives in paper and fabrics to avoid dyes. Perfluorooctane sulfonate and perfluorohexane sulfonate are globally distributed, and contaminates air, water, food, and dust, have toxic effects and bioaccumulate. Significant levels of these compounds have found in blood serum, breast milk, and semen of occupationally exposed and unexposed people, as well as in blood serum and organs of the domestic, farm, and wild animals. The present study seeks to analyze the toxic effects and possible alterations caused by the presence of these compounds in boar sperm during the in vitro capacitation, due to their toxicity, worldwide distribution, and lack of information in spermatozoa physiology during pre-fertilization processes. ABSTRACT: Perfluorooctane sulfonate (PFOS) and perfluorohexane sulfonate (PFHxS) are toxic and bioaccumulative, included in the Stockholm Convention’s list as persistent organic pollutants. Due to their toxicity, worldwide distribution, and lack of information in spermatozoa physiology during pre-fertilization processes, the present study seeks to analyze the toxic effects and possible alterations caused by the presence of these compounds in boar sperm during the in vitro capacitation. The spermatozoa capacitation was performed in supplemented TALP-Hepes media and mean lethal concentration values of 460.55 μM for PFOS, and 1930.60 μM for PFHxS were obtained. Results by chlortetracycline staining showed that intracellular Ca(2+) patterns bound to membrane proteins were scarcely affected by PFOS. The spontaneous acrosome reaction determined by FITC-PNA was significantly reduced by PFOS and slightly increased by PFHxS. Both toxic compounds significantly alter the normal capacitation process from 30 min of exposure. An increase in ROS production was observed by flow cytometry and considerable DNA fragmentation by the comet assay. The immunocytochemistry showed a decrease of tyrosine phosphorylation in proteins of the equatorial and acrosomal zone of the spermatozoa head. In conclusion, PFOS and PFHxS have toxic effects on the sperm, causing mortality and altering vital parameters for proper sperm capacitation.