华东师范大学学报(自然科学版) ›› 2007, Vol. 2007 ›› Issue (4): 86-94.

• 生命科学 • 上一篇    下一篇

保存液及保存条件对中华绒螯蟹精子存活率的影响

陈东华, 周忠良, 范丽君, 曲 迪, 赵云龙, 王 群
  

  1. 华东师范大学 生命科学学院， 上海 200062
  • 收稿日期:2006-02-20 修回日期:2006-04-20 出版日期:2007-07-25 发布日期:2007-07-25
  • 通讯作者: 王群

Viability Rates of Eriocheir sinensis Sperms in SixPreservation Solutions (In Vitro)(Chinese)

CHEN Dong-hua, ZHOU Zhong-liang, FAN Li-jun, QU Di, ZHAO Yun-long, WANG Qun
   

  1. School of Life Science, East China Normal University, Shanghai 200062, China
  • Received:2006-02-20 Revised:2006-04-20 Online:2007-07-25 Published:2007-07-25
  • Contact: WANG Qun

摘要： 以精子存活率和精子密度作为判据，研究了低温（4 ℃）条件下6种保存液体外保存中华绒螯蟹精子的效果.精子存活率采用伊红染色法检测，六种保存液分别为无钙离子人工海水I、无钙离子人工海水II、海水等渗NaCl溶液、无菌中华绒螯蟹生理盐水、海水等渗KCl溶液和海水等渗葡萄糖溶液.结果显示，六种保存液中海水等渗KCl溶液和海水等渗葡萄糖溶液的保存效果最差，保存2 d后精子即全部死亡，而两种无钙离子人工海水的保存效果较好，其中以无钙离子人工海水II的效果尤佳，其保存6 d后的精子存活率达92.19%，精子密度亦保持较高水平；故无钙离子人工海水II是中华绒螯蟹精子短期体外保存的理想保存液.在此基础上以无钙离子人工海水II作保存液，研究了精子密度、保存液pH值和渗透压对保存效果的影响.结果显示，不同精子密度的保存效果差异显著，其中以106～107个/mL的保存密度效果最好，在保存2 d后精子存活率仍保持在95 ％以上；在7~10的范围内保存效果较好，保存15 d后精子存活率均在90%以上；在以NaCl配制的渗透压范围为500～1 000 mOsmol/kg H2O的10个梯度保存液中，渗透压在750～850 mOsmol/kg H2O的范围内保存效果最好，该条件下保存15 d后精子存活率仍在75 ％以上.以上结果证实，中华绒螯蟹精子适宜的保存条件为：精子密度为106～107个/mL，渗透压范围为750～850 mOsmol/kg H2O，pH值为pH 7~10.

关键词: 中华绒螯蟹, 精子, 低温保存, 渗透压, 精子存活率, 精子密度, 中华绒螯蟹, 精子, 低温保存, 渗透压, 精子存活率, 精子密度

Abstract: To evaluate the preservation effects of different solutions, viability rate and density were assessed in of Eriocheir sinensis preserved in six solutions (In Vitro) at 4 ℃. Meanwhile viability rate of sperms was assessed by eosin stain. Those six preservation solutions were calcium-free artificial sea water I (Ca2+-FASW I), calcium-free artificial sea water II (Ca2+-FASW II), isostonie sodium chloride (NaCl) solution, aseptic physiological saline, isostonie potassium chloride (KCl) solution and isostonie glucose solution. Results showed that most of sperms preserved in KCl solution and glucose solution were thorough dead after 2 days. While sperms in Ca 2+-FASW I, II still keep alive.Especially in the Ca2+-FASW II, 92.19% sperms survived after 6 days′ preservation. Therefore, Ca2+-FASW II is an excellent solution to preserve Eriocheir sinensis sperms (In Vitro). Furthermore, Ca2+-FASW II was used to study its preservation effect with different sperms destiny, pH value and osmotic pressure. It indicated that no less than 95% sperms survived after 2 days′ preservation at 106~107 sperm/mL at 4 ℃. At least 90% survived after 15 days′ preservation at pH 7~10. More than 75% survived after 15 days′ preservation at 700~850 mOsmol/kg H2O among 10 scalar solutions at the different osmotic pressures (500~1 000 mOsmol/kg H2O) modulated by NaCl. It is concluded that with sperm density at 106~107 sperm/mL, osmotic pressure at 750~850 mOsmol/kg H2O and pH at 7~10, Eriocheir sinensis sperms are preserved in the optimum condition (In Vitro).

Key words:  spermatozoa, conservation, osmotic pressure, density, Eriocheir sinensis,  spermatozoa, conservation, osmotic pressure, density

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