| 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 |
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| THE VIABILITY RATE OF ERIOCHEIR SINENSIS SPERMS IN VITRO IN PRESERVATION SOLUTIONS OF Ca2+, Mg2+ AND K+ AT DIFFERENT CONCENTRATIONS |
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CHEN Dong-Hua, LI Yan-Dong, JIA Lin-Zhi, LI Xiao-Bo, WANG Li-Ren, SUN Ju-Xiang, WANG Qun
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School of Life Science, East China Normal University, Shanghai, 200062
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| 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+ |
