Localization and expression analysis of sperm-specific glyceraldehyde 3-phosphate dehydrogenase in bull spermatozoa with contrasting sperm motility.

BACKGROUND

Poor sperm motility leading to male infertility has become a profound crisis to be addressed in this contemporary era. In many cases, the origin of poor sperm motility remains unexplained. Few studies reported the indispensable role of sperm-specific glyceraldehyde 3-phosphate dehydrogenase (GAPDHS) in sperm motility, however, studies on GAPDHS are severely confined.

OBJECTIVES

The present study aimed to assess the localization patterns, expression levels, and enzyme activity of GAPDHS in normal and asthenozoospermic bulls and to examine their association with sperm functional parameters.

MATERIALS AND

METHODS

The bull semen samples were classified into high-motile and low-motile groups (n = 7 per each group) based on the ejaculate rejection rate. Sperm kinetic parameters were assessed using computer-assisted sperm analysis (CASA). Sperm viability, mitochondrial membrane potential (MMP), reactive oxygen species (ROS), and intracellular calcium levels were measured through flow cytometry. Subsequently, GAPDHS localization was observed via immunocytochemistry. The expression levels and enzyme activity of GAPDHS were estimated using western blotting and a GAPDHS activity assay kit.

RESULTS AND DISCUSSION: Sperm viability, MMP, ROS, and live sperm intracellular calcium levels did not differ significantly between high and low motile groups. A significant positive correlation was found between MMP and sperm viability, whereas no significant association was found between MMP and sperm progressive motility. The GAPDHS was localized in the principal piece, head-midpiece junction, and at the acrosome region of bull sperm. GAPDHS localization intensity, expression levels, and enzyme activity were found significantly (p < 0.05) higher in the high motile group than in low motile group. Furthermore, we noticed a significant positive correlation between GAPDHS activity and sperm kinetic parameters.

CONCLUSIONS

The analysis of GAPDHS localization patterns, expression levels, and enzyme activity indicated its potential role in sperm motility, suggesting that GAPDHS could serve as a candidate biomarker for sperm motility and male fertility.