Optimizing Gamete Cryopreservation in Saanen Bucks: The Contribution of Vitamin D

Abstract

Vitamin D plays an important role in the reproductive system and the deficiency of it increases the possibility of male infertility. Vitamin D acts in the ATP generation system of the sperm cell by hyperactivating calcium homeostasis. The present study aimed to assess the effects of supplementing various doses of Vitamin D on goat sperm after the cryopreservation process. Semen samples were gathered from six Saanen bucks (2 - 4 years old) at least five times in the breeding season. Cryopreservation was conducted with TRIS - citric acid - lecithin based (1%) - extender in four study groups (Control and 25, 50 and 100 ng/mL Vitamin D supplemented). Sperm samples were frozen in 0.25mL straws. Motility, hypo-osmotic swelling test (HOST), acrosomal (PSA-FITC) and DNA integrities (TUNEL) were investigated with phase-contrast microscope. Mitochondrial membrane potential assessments were conducted by JC/1-PI dual staining via flow-cytometer. Supplementing Vitamin D to goat sperm cryopreservation extenders yielded higher values on sperm parameters like motility, membrane functional integrity and integrity of acrosome results compared to the control group. 25 ng/mL group yielded similar results with control in terms of high mitochondrial membrane potential. DNA integrity values were well preserved after cryopreservation except 100 ng/mL Vitamin D supplemented group. Supplementing cryopreservation extenders with various doses of Vitamin D presented higher results compared to the control group in the study with regards to sperm parameters. These results carry a good potential for further investigations and enlighten the possible success rates of fertilization via assisted reproductive biotechnology methods.

Keywords

• Cryopreservation
• DNA Integrity
• Flow cytometry
• Saanen
• Vitamin D

References

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