Effect of Capacitation on Proteomic Profile and Mitochondrial Parameters of Spermatozoa in Bulls

Sperm capacitation is a critical process for fertilization. This work aims to analyze the effect in vitro capacitation had on the proteome and mitochondrial parameters of bull spermatozoa. Viability, mitochondrial membrane potential (MMP), and reactive oxygen species (mROS) were assessed by flow cytometry in noncapacitated (NC) and in vitro capacitated (IVC) sperm. Proteome was evaluated using SWATH-MS. In vitro capacitation significantly induced a decrease in sperm viability, a high MMP, and an increase in mROS production. Within the group of living spermatozoa, the capacitation significantly induced a decrease in healthy mitochondrial spermatozoa, as well as an increase in mROS production, without affecting the MMP intensity. A total number of 72 differentially abundant proteins were found of which 63 were over-represented in the NC sperm group and 9 in the IVC sperm group. It was observed that many proteins associated with the sperm membrane and acrosome were lost during the capacitation process. For the IVC sperm, the functional enrichment was found in proteins related to the oxidative phosphorylation process. Our results indicate that the capacitation process induces a significant loss of seminal plasma-derived membrane proteins and a significant increase in proteins related with the oxidative phosphorylation (OXPHOS) pathway. Data are available via ProteomeXchange with identifiers PXD056424 and PXD042286.