Proteomic analysis of boar spermatozoa and quantity changes of superoxide dismutase 1, glutathione peroxidase, and peroxiredoxin 5 during epididymal maturation

Abstract

Mammalian spermatozoa and their various proteins undergo various modifications during maturation in the epididymis. To characterize proteins that change in quantity during this maturational process, boar spermatozoa were collected from various regions of the epididymis, and extracts were analyzed by two-dimensional gel electrophoresis (2-DE). A number of proteins were identified as changing in quantity, and MALDI-MS analysis revealed that superoxide dismutase 1 (SOD1) from the acrosomal proteins of spermatozoa, and glutathione peroxidase (GPX) and peroxiredoxin 5 from the membranous fraction increased during the epididymal transit of spermatozoa. These proteins are antioxidants that remove reactive oxygen species (ROS); they are presumed to protect spermatozoa during epididymal transit and storage. Western blot analysis of SOD1, GPX and peroxiredoxin 5 showed that these protein levels increased as the spermatozoa traveled from the caput to the cauda epididymis. Activity analysis showed that total SOD activity also increased. Therefore, we conclude that several antioxidant proteins increase during the transit of boar spermatozoa through the epididymis, ultimately contributing to the maturation and/or survival of sperm.