抗冻剂对日本鳗鲡精子活力及运动时间的影响

The effects of different cryoprotectants on the sperm motility and the movement time of Japanese eel (Anguilla japonica)

利用精子稀释液（0.5%氯化钠＋0.25%氯化钾＋0.25%葡萄糖）分别配制4%、8%、12%、16%、20%、24%的二甲亚砜（DMSO）、甘油（Glycerol）、1,2-丙二醇（1,2-Propanediol）、乙二醇（Ethyleneglycol）四种抗冻剂,对日本鳗鲡精子进行激活处理,观察记录精子在不同浓度抗冻剂中的活力和5种运动（激烈运动、快速运动、慢速运动、摆动、死亡）时间。结果表明：4%-24%的四种抗冻剂都可以激活日本鳗鲡精子,其中二甲亚砜浓度为12%时,精子活力最高,平均可达到78.3%;甘油浓度为8%时,精子活力最高,平均可达到72.8%;12%的1,2-丙二醇和乙二醇激活精子后,精子活力都可以达到最高,分别是47.6%和57.8%。日本鳗鲡精子在8%的二甲亚砜、8%的甘油、8%的1,2-丙二醇和8%的乙二醇中,激烈运动时间均为最长,分别为11.8 s、11.1 s、6.0 s和2.9 s;用8%的甘油、16%的二甲亚砜、16%的1,2-丙二醇和12%的乙二醇激活日本鳗鲡精子后,精子存活时间均最长,分别达到154.6 s、45 s、20 s和34 s。与其它几种抗冻剂相比,8%的甘油能显著延长日本鳗鲡精子的寿命。四种抗冻剂对日本鳗鲡精子都有一定的毒性作用,处理后精子的活力和寿命均降低。

Four kinds of cryoprotectants of 4% ,8% , 12%, 16% ,20% ,24% DMSO （ Dimethyl Sulfoxide） , Glycerol,1.2-Propanediol and Ethyleneglycol have been respectively conducted with sperm diluent （0.5% NaCl ＋ 0.25% KCl ＋ 0.25 % Glu）. By means of using these cryoprotectants to activate the sperm of Anguilla japonica and record their motility as well as movement time. There were five kinds of movement time, namely,drastic movement, rapid movement, slow movement, swag movement and death to be observed and recorded under different concentration. The experimental results revealed that the sperm of Japanese eel could be activated by four kinds of cryoprotectants mentioned above. The sperm motility was the best and the average motility was 78.3% ,72.8% ,47.6% ,57.8% with 12% DMSO,8% Glycerol,12% 1,2-Propanediol and 12% Ethyleneglycol. The drastic movement time of the sperm was the longest and respectively achieved 11.8 s, 11.1 s,6.0 s and 2.9 s with 8% DMSO,8% Glycerol,8% 1,2-Propanediol and 8% Ethyleneglycol respectively. The sperm longevity was the longest and respectively reached 154.6 s,45 s,20 s and 34 s with 8% Glycerol,16% DMSO, 16% 1,2-Propanediol and 12% Ethyleneglycol. As compared with other cryoprotectants, 8% Glycerol could obviously prolong the life of the sperm. Four kinds of cryoprotectants, all of them could produce toxicity on the sperm. The motility and life of the sperm could be reduced after activated by these cryoprotectants.