Exploring Seminal Plasma GSTM3 as a Quality and In Vivo Fertility Biomarker in Pigs—Relationship with Sperm Morphology

Abstract

Glutathione S-transferases Mu 3 (GSTM3) is an essential antioxidant enzyme whose presence in sperm has recently been related to sperm cryotolerance, quality and fertility. However, its role in seminal plasma (SP) as a predictor of the same sperm parameters has never been investigated. Herein, cell biology and proteomic approaches were performed to explore the presence, origin and role of SP-GSTM3 as a sperm quality and in vivo fertility biomarker. GSTM3 in SP was quantified using a commercial Enzyme-Linked Immunosorbent Assay (ELISA) kit specific for Sus scrofa, whereas the presence of GSTM3 in testis, epididymis and accessory sex glands was assessed through immunoblotting analysis. Sperm quality and functionality parameters were evaluated in semen samples at 0 and 72 h of liquid-storage, whereas fertility parameters were recorded over a 12-months as farrowing rate and litter size. The presence and concentration of GSTM3 in SP was established for the first time in mammalian species, predominantly synthesized in the epididymis. The present study also evidenced a relationship between SP-GSTM3 and sperm morphology and suggested it is involved in epididymal maturation rather than in ejaculated sperm physiology. Finally, the data reported herein ruled out the role of this antioxidant enzyme as a quality and in vivo fertility biomarker of pig sperm.

Metabolite Profiling of Pig Seminal Plasma Identifies Potential Biomarkers for Sperm Resilience to Liquid Preservation

“…However, when samples were divided considering breeds, no effect on SP-metabolites concentration was observed, except for fumarate, which exhibited higher levels in Pietrain than in Duroc boars. This agrees with most of the studies evaluating SP components in samples from different pig breeds, in which no breed effect was reported (Barranco et al, 2015b(Barranco et al, , 2016(Barranco et al, , 2020Llavanera et al, 2020). However, since the sample size for each breed was limited in the present study, further research should verify this assumption.…”

“…Many efforts have been focused on exploring pig SP composition with the aim to uncover SP biomarkers able to predict semen quality after ejaculate collection (López Rodríguez et al, 2012;Barranco et al, 2016Barranco et al, , 2017Barranco et al, , 2020Llavanera et al, 2020). In the last years, the advances in metabolomic analytical methods have allowed the identification of SP-metabolites as potential sperm quality biomarkers in mammalian species, such as humans (Bonechi et al, 2015;Engel et al, 2019;Wang et al, 2019;Mumcu et al, 2020;Xu et al, 2020) and cattle (Moura et al, 2018).…”

“…Sperm quality of pig seminal AI-doses gradually declines during their storage at 17 • C (Waberski et al, 2011;Yeste, 2017), showing differences in their sperm resilience to liquid preservation (Parrilla et al, 2012;Zakošek Pipan et al, 2014;Roca et al, 2015). While several studies have demonstrated that some SP-proteins are able to predict pig sperm resilience to liquid storage (Zakošek Pipan et al, 2014;Barranco et al, 2016Barranco et al, , 2019Parrilla et al, 2019;Llavanera et al, 2020), no study has, to the best of our knowledge, evaluated the potential relationship between SP-metabolite composition and the sperm ability to sustain 72 h of liquid storage. Our approach suggested that some SP-metabolites were positively and negatively related with the sperm functionality decline during liquid preservation.…”

Seminal Plasma: Relevant for Fertility?

“…Markers for sperm dysfunction [ 218 ] and (in)fertility have been detected in spermatozoa [ 27 , 190 , 219 , 220 ] and in the SP [ 221 , 222 ] of many species, including humans [ 218 , 222 , 223 , 224 , 225 ] and livestock [ 70 , 226 ]. They vary from gene presence [ 227 ], markers of Sertoli cell function [ 228 ], proteins [ 36 , 229 , 230 , 231 ], anti-oxidative enzymes [ 223 , 232 , 233 ] and metabolites [ 234 , 235 , 236 ] to exosome-associated molecules [ 237 , 238 ] and microRNAs [ 181 , 239 , 240 , 241 ].…”

Protein Identification of Spermatozoa and Seminal Plasma in Bottlenose Dolphin (Tursiops truncatus)

“…Our data also report the presence of this protein in the seminal plasma of dolphins. Recently, it has been demonstrated that low concentration of GSTM3 in pig seminal plasma is related to an increased percentage of sperm abnormalities (Llavanera et al, 2020a). ROPN1 protein is found in sperm flagellum as part of the fibrous sheath, specifically located in the principal piece and end piece of the flagellum (Fujita et al, 2000;Chen et al, 2011).…”