Effect of lactoferrin on ram sperm motility after cryopreservation

Abstract

Objective:The objective of this study was to analyse the differentially abundant proteins caused by freeze-thawing of ram sperm and explore candidate proteins of interest for their ability to improve ram sperm cryopreservation outcomes in vitro .Methods: Sperm were from three mature Dorper. Fresh and frozen sperm proteins were extracted, and the differentially abundant proteins were analysed by mass spectrometry (MS).Among these proteins, lactoferrin(LTF) was selected to be added before cryopreservation.Next, sperm samples were diluted in Tris extender, with the addition of 0μg/ml,10 μg/ml, 100 μg/ml, 500 μg/ml, 1000 μg/ml of LTF. After thawing, sperm quality evaluated by motility, plasma membrane integrity, mitochondrial activity and reactive oxygen species (ROS).Results: Cryopreservation significantly altered the abundance of 40 proteins; the abundance of 16 proteins was increased, while that of 24 proteins was decreased. Next, LTF was added to Tris extender applied to ram sperm. The results showed that sperm motility and plasma membrane integrity were significantly improved (p<0.05) by supplementation with 10 μg/ml LTF compared to those in the control group. There was no significant difference in mitochondrial activity between the 0μg/ml group and other groups (p>0.05). Supplementation of the cryoprotective extender with 10 μg/ml LTF led to decreased reactive oxygen species (ROS) levels compared with those in the control and other groups (p<0.05). Conclusion:LTF is an important protein during cryopreservation, and the addition of 10 μg/ml LTF to a cryoprotective extender can significantly improve the function of frozen ram

Lactoferrin: A glycoprotein that plays an active role in human health

“…Kobayashi et al reported that the addition of Lf can effectively improve sperm activity and prolong the survival time of frozen-thawed semen after thawing, because Lf maintains the integrity of sperm plasma membrane and reduces the level of reactive oxygen species (ROS) (74). This has also been reported in horses (75), goats (76), and porcine (77).…”

Molecular Biomarkers of Canine Reproductive Functions

“…The exact function of LTF in the canine reproductive system is not known, but since it is present in high amounts, it must be important for canine reproductive functions. The addition of LTF to a cryoprotective extender can significantly improve the function of frozen ram sperm [ 66 ].…”

Quality and Protein Profiles in Local Indonesian Ram Sperm Before and After Cryopreservation

“…Such modifications, indicative of biomedical and structural stress experienced by sperm, ultimately result in reduced post-thaw quality (Tamburrino et al 2023). Furthermore, cryopreservation significantly impacts the abundance of various proteins in frozen-thawed ram sperm, with some protein levels increasing while others decrease (Su et al 2022). This process also alters the protein profile of ram sperm during in vitro capacitation, affecting sperm capacitation at the molecular level (Peris-Frau et al 2019).…”