皱纹盘鲍精子的激活及超低温保存研究

Activation and Cryopreservation of Spermatoza in Haliotis discus

用不同浓度的氨海水激活解剖获得的皱纹盘鲍精子并进行激活率和受精率的测定.以自然海水作基础溶液,以二甲基亚砜(DMSO)和甘油(Gly)做保护剂,在不同降温程序下,将解剖获得和经诱导自然排放的皱纹盘鲍精子置于液氮罐中保存24 h,解冻后检测精子的成活率.结果表明,加入自然海水后有64.27%的解剖精子被激活,但不能受精,经氨海水处理后的精子在0.02‰浓度条件下激活率和受精率最高,分别为85.60%、67.42%.自然排放和解剖获得的精子在14%DMSO、4℃平衡15 min,-20℃时平衡5min,-80℃时平衡5 min条件下精子存活率最高,分别为48.15%和64.01%.

In this study, the sperm samples collected from testis anatomy of Haliotis discus are activated using dif- ferent concentrations of ammonia sea water, and the activation rate and fertilization rates are determined. Sperm samples obtained from dissection and natural emission are immersed into anti-freezing agents, DMSO and Gly, through different cooling procedures, and are then immersed in liquid nitrogen for 24 hours. The survival rates are calculated after frozen. The results show that 64.27 % of sperm collected from testis anatomy of abalone can be acti- vated using natural seawater, but can not fertilize the eggs. The highest activation rate of 85.60% and fertilization rate of 61.86 % are observed when the samples are treated with 0.02 ‰ ammonia water before freezing. The high- est survival rates of sperm collected from natural emission and dissection are 48.15 % and 64.01%, respectively, when protected using 14% DMSO, and balanced at 4℃ for 15 min, - 20 ℃ for 5 min, - 80℃ for 5 min.