Extracellular Reactive Oxygen Species (ROS) Production in Fresh Donkey Sperm Exposed to Reductive Stress, Oxidative Stress and NETosis

Jenny shows a large endometrial reaction after semen influx to the uterus with amount of polymorphonuclear neutrophils (PMN) migrating into uterine lumen. PMN act as sperm selection mechanism through phagocytosis and NETosis (DNA extrudes and, together proteins, trap spermatozoa). While reduced percentage spermatozoa are phagocytosed by PMN, most found be attached neutrophil extracellular traps (NETs). This process metabolism produces reactive oxygen species (ROS) that influence reproductive success. The present study aimed determine ROS production in both PMN. With this purpose, (1) donkey were exposed reductive oxidative stresses, adding different concentrations glutathione (GSH) hydrogen peroxide (H2O2), respectively; (2) subjected presence whole semen, sperm, seminal plasma (SP) or other activators such formyl-methionyl-leucyl-phenylalanine (FMLP). Extracellular (measured H2O2 levels) was determined Amplex® Red Hydrogen Peroxide/Peroxidase Assay Kit. Donkey showed more resilience stress than one, GSH treatments led greater production. Moreover, not only did SP appear main inducer but it also able maintain levels produced NETosis.