东北虎精液冷冻及精子超微结构损伤的初步研究  

Preliminary study on cryopreservation of Siberian tiger semen and the ultrastructure damage of the sperm

作　　者：王亮[1] 刘洋 刘丹[3] 马国庆[3] 刘玉堂[1] 

机构地区：[1]东北林业大学野生动物资源学院,黑龙江哈尔滨150040 [2]哈尔滨市剑桥第三中学校,黑龙江哈尔滨150040 [3]黑龙江东北虎林园,黑龙江哈尔滨150028

出　　处：《中国兽医科学》2016年第3期386-392,共7页Chinese Veterinary Science

基　　金：国家林业局野生动植物保护与自然保护区管理司资助(林计发2015)

摘　　要：通过对东北虎林园的东北虎进行电刺激采精的方法取得精液,分别使用葡萄糖和果糖两种稀释液对其进行超低温液氮冷冻,用扫描电镜和透射电镜观察比较冻融前、后精子超微结构的变化。结果显示,与果糖冷冻稀释液相比,葡萄糖稀释液保存的精子损伤程度更大,冻融后精子活率仅达13.83%,而果糖稀释液为58.85%;冷冻对精子的膜和线粒体造成了一定程度的损伤:精子在冻融过程中,质膜易发生膨胀或断裂、顶体可能出现囊泡化或溃散,而质膜和顶体丢失的现象较少;线粒体相互嵌合式连接出现断裂,有的线粒体丢失。上述结果证实,果糖稀释液更适于东北虎精子的保存,虎精子冷冻损伤主要发生在精子膜和线粒体。The ultrastructure of Siberian tiger sperm was studied by scanning electron microscope（SEM） and transmission electron microscope（TEM） before and after freezing, respectively.The semen was collected by electro-ejaculation,and preserved by liquid nitrogen with different cryoprotective diluent containing glucose extender or fructose extender.The results indicated that the damage degree of sperm preserved in glucose diluent is larger than that in fructose one.The post-thaw motility of the former is only 13.83%while the latter is 58.85%.The cell membrane and mitochondria of the sperm suffered a certain degree of damage.In the process of freezing and thawing,the plasma membrane was expanded easily or broken and the acrosome was vesiculated or dispersed while plasma membrane and acrosome seldom lost.The breakage of mitochondrial gomphosis-type connection also appeared during the process,even sometimes accompanying with the loss of mitochondria.In conclusion,the fructose cryoprotective diluent is more applicable to the cryopreservation of Siberian tiger semen,and the cryodamage mainly appears in the cell membrane and mitochondria.

关 键 词：东北虎 精子 冷冻稀释液 超微结构 冷冻损伤 

分 类 号：S864.3[农业科学—野生动物驯养]