引用本文:	陈东华,李艳东,贾林芝,厉小波,王立人,孙菊香,王群.保存液中Ca²⁺、Mg²⁺、K⁺对中华绒螯蟹（Eriocheir sinensis）精子体外保存存活率的影响.海洋与湖沼,2007,38(6):585-589.

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*保存液中Ca²⁺、Mg²⁺、K⁺对中华绒螯蟹（Eriocheir sinensis）精子体外保存存活率的影响*
陈东华, 李艳东, 贾林芝, 厉小波, 王立人, 孙菊香, 王群
华东师范大学生命科学学院上海200062

摘要:

以伊红染色法检测样品精子存活率为依据,研究了4°C下5种保存液及Ca²⁺、Mg²⁺离子对中华绒螯蟹精子体外保存效果的影响。5种保存液分别为人工海水(ASW)、2倍钙离子人工海水(2×Ca²⁺-ASW)、无镁离子人工海水(Mg²⁺-FASW)、无钾离子人工海水(K⁺-FASW)、无钙离子人工海水(Ca²⁺-FASW),经4天保存后,各保存液中精子样品的存活率和精子密度均出现明显差异,K⁺-FASW、ASW及2×Ca²⁺-ASW三种保存液中的精子因发生顶体反应而大量死亡,而Mg²⁺-FASW、Ca²⁺-FASW的保存效果较好。在此基础上,进一步探讨了不同Mg²⁺和Ca²⁺浓度对精子存活率的影响,结果发现,经24h保存后各实验组精子存活率均随着两种离子浓度的增加而明显下降。上述结果表明:K⁺-FASW、ASW及2×Ca²⁺-ASW不适合精子保存,而Mg²⁺-FASW和Ca²⁺-FASW均可作为该蟹精子的保存液;Ca²⁺因可引起精子顶体反应而造成保存液中精子的大量死亡,其浓度与存活率呈明显的负相关;无K⁺的保存液中,Ca²⁺、Mg²⁺的存在与否对精子的保存效果起关键作用;无Mg²⁺人工海水之所以具有较好的保存效果，可能与Mg²⁺的缺乏而导致Ca²⁺逆浓度差转运受阻，避免了因Ca²⁺进入而诱发顶体反应有关。

关键词: 中华绒螯蟹精子低温保存保存液 Ca²⁺ Mg²⁺ K⁺

DOI：

分类号:

基金项目:国家自然科学基金资助项目,30300265号

附件

THE VIABILITY RATE OF ERIOCHEIR SINENSIS SPERMS IN VITRO IN PRESERVATION SOLUTIONS OF Ca²⁺, Mg²⁺ AND K⁺ AT DIFFERENT CONCENTRATIONS

CHEN Dong-Hua, LI Yan-Dong, JIA Lin-Zhi, LI Xiao-Bo, WANG Li-Ren, SUN Ju-Xiang, WANG Qun

School of Life Science, East China Normal University, Shanghai, 200062

Abstract:

The viability rate of in vitro sperms of Eriocheir sinensis preserved in five solutions at 4°C was studied with eosin stain to evaluate the impact from metal ions. The five preservation solutions (Tab.1) included artificial seawater (ASW), 2-fold calcium ion artificial seawater (2×Ca²⁺-ASW), magnesium-free artificial seawater (Mg²⁺-FASW), potassium-free artificial seawater (K⁺-FASW) and calcium-free artificial seawater (Ca²⁺-FASW). Results show that the sperms preserved in K⁺-FASW, ASW, and 2×Ca²⁺-ASW were completely killed 4 days later because of acrosome reaction. However, most of the sperms in Mg²⁺-FASW and Ca²⁺-FASW remained alive in the same period. Therefore, Mg²⁺-FASW and Ca²⁺-FASW can be ideal solutions for preserving E. sinensis sperms in vitro, but K⁺-FASW, ASW, and 2×Ca²⁺-ASW. Furthermore, the viability rate was also checked with different concentrations of Ca²⁺ and Mg²⁺. As the concentrations of Ca²⁺ and Mg²⁺ incremented, the viability rate dropped in 24 hour, indicating that Ca²⁺ could lead to mass mortality of the sperms due to acrosome reaction. Inverse correlation was found between the viability rate and Ca²⁺ concentration. Ca²⁺ and Mg²⁺ played a key role in e K⁺-free preservation solutions. The good performance of Mg²⁺-FASW is probably due to the prohibition of Ca²⁺ transportation against the concentration difference, so that Ca²⁺ was kept away from entering the sperms resulting in acrosome reaction.

Key words: Eriocheir sinensis, Spermatozoa, Low temperature preservation, Preservation solution, Ca²⁺, Mg²⁺, K⁺