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.
Highlights
Artificial insemination (AI) is one of the major breakthroughs of pig reproductive biotechnology and has become the main technique for the breeding of this species worldwide, being an essential tool to achieve productivity challenges in swine industry [1]
It is to suggest that exploring seminal plasma (SP)-Glutathione S-transferases Mu 3 (GSTM3) as a sperm quality and in vivo fertility biomarker may improve reasonable to suggest that exploring SP-GSTM3 as a sperm quality and in vivo fertility biomarker the evaluation of reproductive performance of pig AI-doses
To the best of our first report confirming the presence of GSTM3 in SP of mammals, which underpins the contribution of knowledge, this is the first report confirming the presence of GSTM3 in SP of mammals, which testis, epididymis and accessory sexual glands to GSTM3 content in SP and assessing the putative role underpins the contribution of testis, epididymis and accessory sexual glands to GSTM3 content in SP
Summary
Artificial insemination (AI) is one of the major breakthroughs of pig reproductive biotechnology and has become the main technique for the breeding of this species worldwide, being an essential tool to achieve productivity challenges in swine industry [1]. AI can be performed using both frozen-thawed and liquid-stored sperm at 17 ◦ C, the latter is used in the vast majority of pig (i.e., motility, morphology and plasma membrane integrity), farrowing rates are often suboptimal after liquid-storage [3,4]. In this regard, it is estimated that about 6% of spermiogram-normal AI-pigs are subfertile individuals that remain “hidden,” which could lead to reproductive and economic. Identification and quantification of differentially expressed proteins is known as comparative proteomics The application of this emerging approach for the identification of novel SP quality and fertility biomarkers is currently flourishing [9]. Recent studies uncovered the role of antioxidant enzymes in SP, such as glutathione peroxidase 5 (GPX5) and paraoxonase 1 (PON1), as sperm cryotolerance, quality and/or fertility biomarkers [14,15,16]
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