Abstract

Conventional semen analyses are used to evaluate male factor fertility/infertility in humans and other animals. However, their clinical value remains controversial. Therefore, new tools that more accurately assess male fertility based on sperm function and fertilization mechanism are of interest worldwide. While protein markers in spermatozoa that might help differentiate fertile and infertile sperm have been identified, studies are in their infancy, and the markers require validation in field trials. In the present study, to discover more sensitive biomarkers in spermatozoa for predicting male fertility, we assessed protein expression in capacitated spermatozoa. The results demonstrated that cytochrome b-c1 complex subunit 2 (UQCRC2) was abundantly expressed in high-litter size spermatozoa (>3-fold). On the other hand, equatorin, beta-tubulin, cytochrome b-c1 complex subunit 1 (UQCRC1), speriolin, Ras-related protein Rab-2A (RAB2A), spermadhesin AQN-3, and seminal plasma sperm motility inhibitor were abundantly expressed in low-litter size spermatozoa (>3-fold). Moreover, RAB2A and UQCRC1 expression negatively correlated with litter size, while UQCRC2 expression positively correlated with litter size. Finally, the putative biomarkers predicted litter size in field trials. Our study suggests that biomarkers present in spermatozoa after capacitation can help differentiate superior male fertility from below-average fertility with high sensitivity.

Highlights

  • Conventional semen analyses are used to evaluate male factor fertility/infertility in humans and other animals

  • Equatorin, beta-tubulin, cytochrome b-c1 complex subunit 1 (UQCRC1), speriolin, Ras-related protein Rab-2A (RAB2A), spermadhesin AQN-3, and seminal plasma sperm motility inhibitor were abundantly expressed in low-litter size spermatozoa (>3-fold)

  • Sperm motion kinematics and morphological changes related to capacitation were evaluated with computer-assisted sperm analysis (CASA) and H33258/CTC staining

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Summary

Introduction

Conventional semen analyses are used to evaluate male factor fertility/infertility in humans and other animals. Semen analyses, such as the sperm morphology test[6], motility test[7], swelling/eosin test[8] and penetration assay[9], have been developed for use in humans and other animals. These tools provide initial quantitative information on semen, their clinical value in predicting fertility is debated[10]. To develop highly sensitive biomarkers for the evaluation of male fertility, comprehensive studies of capacitated spermatozoa that yield high- and low-litter sizes in boar were performed. Differential protein expression was confirmed with western blotting, and enzyme-linked immunosorbent assay (ELISA) was used to determine the relationship between protein expression at capacitation and fertility

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