Effect of a novel hydrogen sulfide donor on sperm physiology

Bc. Marie Ladová

Master's thesis

Abstract:

Hydrogen sulfide (H2S) donors are essential tools in exploring the physiological role of H2S in cell function. A wide range of H2S donors is available that mimics physiological H2S levels under in vitro conditions. Despite the significant advances in H2S research, the effects of these compounds on sperm function are not fully understood. This diploma thesis aimed to investigate the effect of a novel N-thiocarboxyanhydride (NTA) derived from the amino acid valine (Val-NTA), which releases H2S in the presence of the ubiquitous enzyme carbonic anhydrase, on porcine sperm quality during semen storage. Specifically, sperm motility, kinetics, plasma membrane integrity, and acrosome integrity were evaluated in porcine sperm samples with or without NTA supplementation at 0 h (control group only), 17 h, and 45 h of semen storage at 17 °C. The application of Val-NTA did not exhibit any positive influence on the sperm plasma membrane integrity, acrosomal integrity, motility, or kinetics. Additionally, the findings from this study demonstrated that Val-NTA exhibited dose-dependent toxicity towards spermatozoa, with specific concentrations resulting in a significant decrease in several sperm parameters. This was consistent with previous studies, which found that high concentrations of H2S donors could have negative effects on sperm motility and induce oxidative stress. Specifically, Val-NTA at the concentration of 100 μM was found to cause a drastic reduction in most of the sperm parameters evaluated. The results also showed a decline in the sperm parameters during storage, which is consistent with previous research and might at least partly be linked to oxidative stress that can be provoked by the exposure of cells to low temperatures. The study suggests that different environmental conditions (the length of incubation, temperature, oxidative stress or variation of carbonic anhydrase levels in the porcine ejaculates) may have influenced the impact of Val-NTA on sperm plasma membrane integrity, acrosomal integrity, motility, and kinetics and that further research, specifically of the mitochondrial activity, is warranted to determine the potential of Val-NTA.

Keywords

boar spermatozoa, gasotransmitter, H2S donor, N-thiocarboxyanhydrides, sperm storage

Language used: English

Date on which the thesis was submitted / produced: 17. 4. 2023

Thesis defence

• Date of defence: 15. 5. 2023
• Supervisor: Eliana Pintus, Ph.D.
• Reader: Nicole Sjahrirová, externi

Czech University of Life Sciences Prague

Faculty of Agrobiology, Food and Natural Resources

Master programme / specializace:
Biotechnology / Reproduction Biotechnology