摘要
采用具有高繁殖力的种公猪的新鲜精液及冷冻精液，加入不同获能剂咖啡因及透明质酸，在３９．５℃、５％ＣＯ２培养箱中孵化培养。结果表明，透明质酸能够在体外条件下保持精子的活率，并维持在稳定的水平，鲜精为４２．６％～４７．８％，冻精为２０．４％～２５．６％。荧光剂Ｈｏｅｃｈｓｔ染色表明，透明质酸能够显著增加培养３６０ｍｉｎ后的活精子率。在无蛋白来源的ｍＢＯ培养液中，培养０～６０ｍｉｎ，精子发生自发的获能和顶体反应，而受获能剂影响，精子发生获能的时间带在各处理组之间显著不同：咖啡因处理组的获能时间带是６０～３６０ｍｉｎ，透明质酸处理组则是６０～１８０ｍｉｎ。体外受精试验表明，去除卵母细胞周围卵丘细胞时，透明质酸和咖啡因对精子穿透率的影响没有区别；而在卵丘细胞存在的前提下，咖啡因处理组的穿透率显著高于透明质酸处理组。
The frozenthawed and fresh spermatozoa were used in this experiment from the proven fertility boars. Caffeine and hyaluronic acid were added as capacitation reagent and incubated with spermatozoa in 39.5℃ and 5% CO2 incubator. At different stages of incubation, motility was examined under a light microscope; viable, capacitated and acrosomal reacting spermatozoa were counted by chlortetracycline staining. Frozenthawed spermatozoa was also used to fertilize oocytes matured in vitro. The results showed that hyaluronic acid could keep the motility at 42.647.8% in fresh and 20.425.6% in frozenthawed spermatozoa. Motility of spermatozoa in hyaluronic acid treated group didn′t change within 6 hr of incubation. In contrast, motility in caffeine treated group showed a peak in 3060 min of incubation, and then decreased during further incubation. Motility in control group (without any treatment) decreased gradually. Hoechst staining experiment showed that hyaluronic acid could improve the viability at 360 min of incubation, whereas, spermatozoa death rate in caffeine treated and control group increased significantly at 180360 min of incubation. In defined mBO, spermatozoa capacitated and underwent acrosomal reaction spontaneously during 060 min of incubation; and the capacitation duration was influenced by capacitation reagents: caffeine treated group occurred in 60360 min of incubation; hyaluronic acid treated group occurred in 60180 min of incubation. Accordingly, acrosomal reaction initiated by caffeine occurred in 60360 min of incubation, whereas hyaluronic acid inhibited the acrosomal reaction. In vitro fertilization showed that when cumulus cells removed from the oocytecumulus complexes, hyaluronic acid and caffeine had the same effects on penetration rate; when cumulus cells remained, caffeine had a better effect than hyaluronic acid on penetration rate.
出处
《中国兽医学报》
CAS
CSCD
北大核心
1997年第6期600-605,共6页
Chinese Journal of Veterinary Science
基金
瑞典皇家科学院赞助
关键词
猪
精子
活率
活精子率
顶体反应
体外受精
pig
spermatozoa
motility
viability
acrosomal reaction
in vitro fertilization
