Metabolomic Analysis and Identification of Sperm Freezability-Related Metabolites in Boar Seminal Plasma

SIMPLE SUMMARY: In the freezing process of boar sperm, there are obvious differences in freezability between individuals. Studies suggest that specific freezability markers might be useful in good (GFE) and poor freezability ejaculate (PFE) selection prior to cryopreservation. Therefore, we performed UHPLC-qTOF-MS analysis to explore the difference in the metabolic level of seminal plasma between boars with differential freezability, and the results showed that the content of D-aspartic acid, N-acetyl-L-glutamate (NAG), and inosine were significantly different. These findings present new insights into the role of metabolism in sperm freezability and provide research directions for exploring potential biomarkers of freezability. ABSTRACT: Some potential markers of boar sperm freezability have been found in spermatozoa, but little attention has been paid to seminal plasma. The seminal plasma is composed of secretions from the testis, epididymis, and accessory sex glands. The exposure of spermatozoa to small molecules such as metabolites can affect sperm function. However, details and significance of the seminal plasma metabolome related to boar sperm freezability are unknown. Therefore, the main aim of this study was to explore the differences in the metabolic level of seminal plasma between boars with differential freezability and to explore the candidate biomarkers of semen freezability. A total of 953 metabolites were identified in boar semen plasma by UHPLC-qTOF-MS analysis, and 50 metabolites showed significant change between the GFE group and PFE group. Further, twelve metabolites were subjected to metabolic target analysis, and three metabolites (D-aspartic acid, N-acetyl-L-glutamate (NAG), and inosine) showed differences. In conclusion, there is significant difference in the metabolome of seminal plasma between GFE and PFE individuals. D-aspartic acid, NAG, and inosine in seminal plasma may be potential markers for assessing sperm cryopreservation resistance in boars.