Mitochonic Acid 5 Increases Ram Sperm Quality by Improving Mitochondrial Function during Storage at 4 °C
by
,
Ruyuan Wang
1,
Luwei Liu
1,
Lingjiang Min
1,
Adedeji O. Adetunji
2,
Xin Kou
3,
Kaifeng Zhou
4 and
Zhendong Zhu
1,* 1
College of Animal Science and Technology, Qingdao Agricultural University, No. 700 Changcheng Road, Qingdao 266109, China
2
Department of Agriculture, University of Arkansas at Pine Bluff, Pine Bluff, AR 71601, USA
3
Hongde Livestock Farm, Yingli Town, Shuoguang 262717, China
4
Shandong Provincial Animal Husbandry General Station, Jinan 250022, China
*
Author to whom correspondence should be addressed.
Animals 2024, 14(3), 368; https://doi.org/10.3390/ani14030368
Submission received: 3 January 2024
/
Revised: 19 January 2024
/
Accepted: 22 January 2024
/
Published: 23 January 2024
(This article belongs to the Special Issue Reproductive Physiology of Ruminants)
Simple Summary
This study investigated the efficacy of mitochonic acid 5 (MA-5) supplementations in enhancing ram sperm quality during 4 °C storage. Cold shocks, osmotic stresses, and oxidative stresses during storage can lead to irreparable damage. Ram sperm diluted with a tris-citrate-glucose extender containing varied MA-5 concentrations were stored for different durations. Notably, the addition of MA-5, particularly at 10 nM, significantly improved sperm quality by enhancing motility, membrane integrity, and acrosomal integrity. MA-5 also increased mitochondrial membrane potential, reduced reactive oxygen species (ROS) levels, and elevated adenosine triphosphate (ATP) levels in ram sperm. The findings suggest that MA-5 preserves high mitochondrial function during liquid storage, providing a promising avenue for further research in ram sperm preservation and specific proteins.
Abstract
Semen preservation involves lengthening sperm’s fertile lifespan without any detrimental effects on its biochemical, functional, and ultrastructural properties. Liquid storage at 4 °C is a ram sperm preservation method. However, this method of storage causes irreversible damage due to cold shocks, osmotic stresses, oxidative stresses, and reductions in sperm metabolism. The present study aims to investigate whether the supplementation of mitochonic acid 5 (MA-5) in a sperm extender could improve chilled ram sperm quality and elucidate its mechanism of action. Ram sperm were diluted with a tris-citrate-glucose extender containing different concentrations of MA-5 (0, 0.1, 1, 10, and 100 nM) and stored at 4 °C for up to 48 h. Sperm motility, membrane integrity, acrosome integrity, mitochondrial membrane potential, reactive oxygen species (ROS) level, ATP content, and the expression of NADPH dehydrogenase subunits 1 (MT-ND1) and NADPH dehydrogenase subunits 6 (MT-ND6) were evaluated. It was observed that compared to the control, the 10 nM MA-5 treatment significantly (p < 0.05) increased total motility (82 ± 3.5% vs. 76 ± 5.9%), progressive motility (67.6 ± 8.2% vs. 51 ± 8.3%), and other parameters (straight-line velocity (VSL), average path velocity (VAP), and curvilinear velocity (VCL)). In addition, 10 nM MA-5 supplementation also improved ram sperm membrane integrity and acrosomal integrity as well increased mitochondrial membrane potential (51.1 ± 0.7% vs. 37.7 ± 1.3%), reduced ROS levels, and elevated adenosine triphosphate (ATP) contents. Furthermore, a Western blot analysis demonstrated that the addition of MA-5 significantly (p < 0.05) increased the expression of MT-ND1 and MT-ND6 proteins in ram sperm, with the 10 nM MA-5 treatment resulting in the highest expression level. These results suggest that MA-5 improves ram sperm quality by maintaining high sperm mitochondrial function during liquid storage at 4 °C.
Keywords:
liquid storage; mitochondria; mitochonic acid 5; ram sperm
