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李新红, 顾孝连, 华修国, 邹兴淮. 2003: 甘油浓度对蓝狐精子深低温冷冻保存的影响及冻融前后精子的超微结构. 动物学研究, 24(6): 421-428.
引用本文: 李新红, 顾孝连, 华修国, 邹兴淮. 2003: 甘油浓度对蓝狐精子深低温冷冻保存的影响及冻融前后精子的超微结构. 动物学研究, 24(6): 421-428.
LI Xin-hong, GU Xiao-lian, HUA Xiu-guo, ZOU Xing-huai. 2003. Effects of Glycerol Concentration on Semen Cryopreservation in Blue Foxes and Ultrastructure of Sperma tozoa Subjected to the Process of Freezing-thawing. Zoological Research, 24(6): 421-428.
Citation: LI Xin-hong, GU Xiao-lian, HUA Xiu-guo, ZOU Xing-huai. 2003. Effects of Glycerol Concentration on Semen Cryopreservation in Blue Foxes and Ultrastructure of Sperma tozoa Subjected to the Process of Freezing-thawing. Zoological Research, 24(6): 421-428.

甘油浓度对蓝狐精子深低温冷冻保存的影响及冻融前后精子的超微结构

Effects of Glycerol Concentration on Semen Cryopreservation in Blue Foxes and Ultrastructure of Sperma tozoa Subjected to the Process of Freezing-thawing

  • 摘要: 人工采取8只优质芬兰雄性蓝狐的精液,分别利用2%、4%、6%和8%甘油浓度的卵黄-Tris-果糖-柠檬酸钠稀释液进行稀释,制成细管冻精。在冻融后0、0.5、2、4、6 h检测4种浓度组的精子运动度、质膜完整率、顶体完整率;并利用透射电镜观察冻融前后精子的超微结构变化。冻融后0 h,4%甘油浓度组冻融精子的运动度、质膜完整率、顶体的完整率均最高(分别为41.8%、43.6%、48.4%),2%浓度组最低(分别为24.5%、27.6%、31.7%);随着检测时间延长,2%与4%组的精子特性差异显著,但2%、6%、8%3个组间差异不显著;6 h时各组间精子的运动度均不超过10%,最高质膜完整率和顶体完整率分别为11.8%、12.7%。说明蓝狐精液稀释剂中甘油的适宜浓度应为4%,冻融后精子的活力维持时间较短。蓝狐精子冻融过程中质膜极易发生膨胀或断裂、顶体囊泡化或溃散,而质膜和顶体丢失现象较少。

     

    Abstract: Eight high quality male blue foxes from Finland were chosen for semen collection by digital manipulation and diluted with the extender Ⅰ.The diluted sperm were divided into four fractions and the same volume of the extender Ⅱ containing glycerol was added to reach the final glycerol concentration of 2%,4%,6%,8% (v/v) ,respectively.After equilibrated for 2 h at 5 ℃.Semen samples were packed in 0.5 mL plastic straws and frozen on a rack 5 cm above liquid nitrogen and thawed at 70 ℃ for 9 s.Before and after freezing-thawing,not only the effects of different concentrations of glycerol on changes of spermatozoa characteristics but also the alterations of ultrastructure of spermatozoa by Transmission Electron Microscope were studied.The results showed,the motility and plasma membrane integrity of spermatozoa together with the percentage of intact acrosomes were related to the dose of glycerol.All these characteristics reached the highest in the fraction that the final glycerol concentration was 4% (41.8%,43.6%,48.4%) and the lowest appeared in the fraction of 2% (24.5%,27.6%,31.7%).Along with the extension of incubation,characteristics of spermatozoa in both fractions of 2% and 4% were significant differences while the differences between 2%,6% and 8% were insignificant.After 6 hours,the motility of each fraction was not more than 10% while the highest viability and the percentage of intact acrosomes were 11.8% and 12.7% respectively.The optimum concentration of glycerol was 4% in the extenders of freezing spermatozoa for blue fox.The living time of spermatozoa subjected to the process of freezing and thawing was short.The plasma membrane was expanded extremely or broken and the acrosome was vesiculated or dispersed while plasma membrane and acrosome seldom lost.

     

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