Seminal plasma protein profiles and testosterone levels as biomarker semen quality of candidate Madura bulls

Abstract

Objective: This study aims to determine the protein profile based on molecular weight (MW) and testosterone levels in seminal plasma (SP) that correlates to the semen quality of candidate Madura bulls. Material and Methods: A total of 10 male candidate madura bulls underwent semen evaluation (motility, viability, membrane plasma integrity (MPI), and sperm concentration). The centrifuge was run at 1,200 rpm (4°C) for 20 min to collect SP. SP testosterone levels were measured using an Enzyme-linked immunosorbent assay. The characterization of SP proteins in Madura bulls was done using 1D sodium dodecyl-sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis. All parameters were analyzed using Pearson correlation analysis. Result: The results of the SDS-PAGE analysis found eight protein bands with the highest MW of 110 kDa and the lowest of 12 kDa. The mean and SD of SP testosterone levels were 20.58 ± 8.56 ng/ml, motility 59.32% ± 20.14%, viability 67.45% ± 20.22%, MPI 32.77% ± 16.52%, and sperm concentration 1,002.64 ± 429.33 106/mm3. Proteins with MWs of 110 and 91 kDa significantly correlated with MPI, and 110 kDa negatively correlated with sperm concentration (p < 0.05). Proteins with MWs of 73 and 36 kDa significantly correlated with SP testosterone levels, while proteins with MWs of 29 kDa significantly correlated with sperm viability (p < 0.05). Conclusion: The expressed protein fraction based on MW is closely related to the quality of semen, so it has the potential to be a biomarker of semen quality. Further research is needed to determine the specific proteins in certain fractions.

Semen quality and frozen semen production in Pasundan bulls: A molecular weight perspective on seminal plasma and spermatozoa protein

“…The low motility and IPM of spermatozoa in the fresh semen of Pasundan Bagaskara bulls could be attributed to the absence of proteins with MWs of 35–50 kDa in the seminal plasma. This result agrees with the finding reported by Azizah et al [21] that the region of proteins with a MW of 45 kDa is positively correlated with motility and IPM of spermatozoa in Madura bull. Moreover, the non-expression of proteins with a MW of 35–50 kD may be associated with glutathione peroxidase 3 (GPX3), which could be associated with low spermatozoa viability.…”

“…The interaction between GPX3 and sulfhydryl oxidase 1 (QSOX1) plays a vital role in how sperm responds to oxidative stress by involving cyclic GMP (cGMP). Cyclic GMP serves as an intracellular component that can modulate nitric oxide and natriuretic peptides, regulating various biological processes [21] and contributing to the maintenance of intact spermatozoa plasma membranes. The low viability and IPM (Bagaskara and Kartarajasa males) are associated with sperm plasma membrane damage due to the generation of oxidative stress through the cGMP mechanism.…”

Quality and Protein Profiles in Local Indonesian Ram Sperm Before and After Cryopreservation

“…Spermatozoa undergo various stresses during cryopreservation, including ice crystal formation, cold shock, chemical poisoning, oxidative stress, and osmotic pressure, all of which can lead to structural damage to the spermatozoa, including alterations to protein molecules. Research by Azizah et al (2023) illustrates the close relationship between semen proteins and traits, with specific proteins associated with freezeability (Rego et al 2016). Developing strategies to ensure high post-thaw sperm quality requires an understanding of how cryopreservation-induced stress impacts sperm biology (Ďuračka et al 2023).…”