Proteomic analysis of spermatozoa reveals caseins play a pivotal role in preventing short-term periods of subfertility in stallions†.

Stallions experience transient fluctuations in fertility throughout the breeding season. Considering pregnancy diagnoses cannot be ascertained until ~14 days postbreeding, the timely detection of decreases in stallion fertility would enhance industry economic and welfare outcomes. Therefore, this study aimed to identify the proteomic signatures reflective of short-term fertility fluctuations and to determine the biological mechanisms governing such differences. Using liquid chromatography-mass spectrometry (LC-MS/MS), we compared the proteomic profile of semen samples collected from commercially "fertile" stallions, during high- and low-fertility periods. A total of 1702 proteins were identified, of which, 38 showed a significant change in abundance (P ≤ 0.05). Assessment of intra- and interstallion variability revealed that caseins (namely κ-, α-S1-, and α-S2-casein) were significantly more abundant during "high-fertility" periods, while several epididymal, and seminal plasma proteins (chiefly, epididymal sperm binding protein 1 [ELSPbP1], horse seminal plasma protein 1 [HSP-1], and clusterin), were significantly more abundant during "low-fertility" periods. We hypothesized that an increased abundance of caseins offers greater protection from potentially harmful seminal plasma proteins, thereby preserving cell functionality and fertility. In vitro exposure of spermatozoa to casein resulted in decreased levels of lipid scrambling (Merocyanine 540), higher abundance of sperm-bound caseins (α-S1-, α-S2-, and κ-casein), and lower abundance of sperm-bound HSP-1 (P ≤ 0.05). This study demonstrates key pathways governing short-term fertility fluctuations in the stallion, thereby providing a platform to develop robust, fertility assessment strategies into the future.

Functions and effects of reactive oxygen species in male fertility.

Reactive oxygen species (ROS) are implicated in all aspects of cellular functions. While the importance of ROS as signalling molecules is well described, ROS are also associated with stress pathologies. Within the reproduction field, there are associations with reduced fertility as a result of lipid peroxidation, protein dysfunction, premature cell death and DNA damage which readily occur in spermatozoa. These oxidative insults can arise in vivo, or in vitro as a result of sperm storage, purification and processing. Following a brief description of the production, homeostasis and functions of ROS in mammalian sperm function, this review paper will focus on describing the predominant sources of ROS in the ejaculate, the effects of ROS on a cellular and molecular level, and the actions of ROS from the whole animal perspective. There is highlighting of some studies, which have revealed the mechanisms for these observations, along with some strategies to ameliorate or prevent the instigation of the oxidative stress cascade before irreversible damage to spermatozoa occurs.