草食家畜Issue(4)：1-9,9.
TRIS和HEPES缓冲体系对驴精液冻融前后效果的影响
The Effect of TRIS and HEPES Buffer Systems on Donkey Semen Before and after Freezing and Thawing
摘要
Abstract
[Objective]To screen the effects of TRIS and HEPES buffer systems on the preservation of donkey semen,this experiment collected semen from 10 healthy male donkeys aged 10-12 years.By comparing the effects of TRIS and HEPES buffer combinations on the low-temperature preservation and freeze-thawing of donkey semen,a more suitable buffer system for the dilution solution was designed for the low-temperature preservation and freeze-thawing of donkey semen.[Methods]A TRIS buffer system composed of 200 mM TRIS and 2 mM citrate salt was added to the donkey semen dilution solution and freezing solution,and a HEPES buffer system composed of 40 mM HEPES and 2 mM citrate salt was also added.Sperm were loaded,equilibrated,and frozen in 0.5 mL cryotubes.The frozen donkey sperm cryotubes were then thawed in a 42 °C water bath for 20 seconds.The two buffer systems were compared by analyzing semen vitality parameters(including total motility TM,progressive motility,etc.),plasma membrane integrity rate,DNA fragmentation rate,and sperm deformity rate.[Results]The HEPES buffer system significantly outperformed the TRIS buffer system in terms of sperm motility parameters before and after freezing and thawing,with extremely significant differences(P＜0.01).Specifically,the overall motility and progressive motility of sperm in the HEPES buffer system were higher than those in the TRIS buffer system.After thawing,the DNA fragmentation rate of sperm in the HEPES buffer system was also significantly better than that in the TRIS buffer system(P＜0.01).There were no significant differences between the TRIS buffer system and the HEPES buffer system in terms of membrane integrity rate and sperm deformity rate(P＞0.05).[Conclusion]HEPES,as a zwitterionic buffer,is more effective than TRIS in donkey semen cryopreservation diluents.The HEPES buffer system significantly improves sperm vitality parameters before and after freezing and thawing and reduces DNA fragmentation rates,which is of great significance for improving the low-temperature storage and freezing-thawing effects of donkey semen.Therefore,HEPES is recommended as the buffer agent in the dilution solution formulation for low-temperature storage and freezing-thawing of donkey semen.关键词
驴/精液/缓冲体系/活力参数/低渗肿胀/DNA碎片化率/畸形率Key words
donkey/semen/buffer system/vitality parameter/hypotonic swelling/DNA fragmentation/deformity rate分类
农业科技引用本文复制引用
郭佳丽,阿依努尔·亚森,罗永明,王中禹,玉山江,齐·阿拉达尔..TRIS和HEPES缓冲体系对驴精液冻融前后效果的影响[J].草食家畜,2025,(4):1-9,9.基金项目
新疆维吾尔自治区重大科技专项"草食家畜高效繁殖技术研究"(2023A2011-3) （2023A2011-3）
