计算机辅助对几种鲟鱼冻精激活液的比较  

Comparative study of activating mediums for the cryopreserved sperm of several sturgeons using CASA

作　　者：柳凌[1] OTOMAR Linhart 危起伟[1] 郭峰[1] 张洁明[1] 刘鉴毅[1] 

机构地区：[1]中国水产科学研究院长江水产研究所农业部淡水鱼类种质资源与生物技术重点实验室 [2]Joint Laboratory of Genetics,Physiology and Reproduction of Fish,Institute of Animal Physiology and Genetics,Academy of Sciences of the Czech Republic,University of South Bohemia,Research Institute of Fish Culture and Hydrobiology,38925 Vodnany,Czech Republic

出　　处：《水产学报》2007年第6期711-720,共10页Journal of Fisheries of China

基　　金：国家基础公益项目(2004DIB3J099)

摘　　要：首次在我国采用鱼类彩色精子图文分析系统(FSQAS-2000)对中华鲟、史氏鲟、西伯利亚鲟和匙吻鲟经超低温冷冻保存后的精子,在不同激活液条件下,平均曲线运动速度(VCL)、平均直线运动速度(VSL)、平均路径运动速度(VAP)、平均移动角度(MAD)、以及冷冻前后精子活率的变化等多项参数进行了统计学比较研究。研究分析了不同渗透压、pH以及含Mg^(2+)的激活液对这些参数的影响。研究结果表明:几种鲟鱼激活液的渗透压均较低,其中西伯利亚鲟最低,为10 mOsm·kg^(-1);中华鲟和史氏鲟为20 mOsm·kg^(-1);匙吻鲟最高,为30 mOsm·kg^(-1)。不同的渗透压对冷冻精子的VCL、VSL、VAP、以及精子活率均产生显著(P≤0.05)或极显著(P≤0.01)影响。激活液的pH除了对冷冻精子的上述参数产生影响外,最显著的是对精子的运动轨迹,即平均移动角度(MAD°·s^(-1))产生影响(P≤0.01)。pH越低,精子的MAD越小。几种鲟鱼的最适pH分别为西伯利亚鲟pH 7.5,中华鲟、史氏鲟和匙吻鲟pH 8.5。当4种鲟鱼的冷冻精子激活液中加入5 mmol·L^(-1)MgCl_2时,冷冻精子的3种运动速度显著提高(P≤0.01)。综合分析后认为:西伯利亚鲟的冷冻精子激活液应为10 mmol·L^(-1)Tris-HCl+5 mmol·L^(-1)MgCl_2,pH 7.5;中华鲟和史氏鲟的冷冻精子激活液为20 mmol·L^(-1)Tris-HCl+5 mmol·L^(-1)MgCl_2,pH 8.5;匙吻鲟的冷冻精子激活液为30 mmol·L^(-1)Tris-HCl+5 mmol·L^(-1) MgCl_2,pH 8.5。用此激活液对冷冻前后4种鲟鱼精子活力的相关性进行线性回归分析发现,西伯利亚鲟的R^2=0.8296(P<0.01);中华鲟的R^2=0.9860(P<0.01);史氏鲟的R^2= 0.9622(P<0.01);匙吻鲟R^2=0.9477(P<0.01)。说明冷冻前后4种鲟鱼精子的平均活率存在着极显著的线性正相关性。The digital analysis technique of sperm quality assessment in fish has provided reliable reference evidences for the effects and the damage mechanism of cryopreservation, genetics and breeding, etc. From the study using fish color sperm image analysis systems （FSQAS-2000） conducted on the cryopreserved sperm of Chinese sturgeon （Acipenser sinensis ）, Amur sturgeon （A. schrenckii）, Siberian sturgeon （A. baerii ） and paddlefish（Polyodon spathula） under different activating mediums, the most parameters, including the curvilinear velocity （ VCL）, the straight line velocity （ VSL）, the angular path velocity （ VAP）, angular displacement （MAD） and percentage of motile sperm, etc. were compared for the first time in China. The influences of activating mediums with different osmolality, pH and concentration of Mg^2 ＋ on the parameters were also analyzed in the study. Our results indicated that： First, the osmolality of activating medium for the sturgeons mentioned above was quite low, among them, the lowest for Siberian sturgeon （10 mOsm· kg^ -1 ）, the highest for paddlefish （ 30 mOsm· kg^ -1 ）, the middle for Chinese sturgeon and Amur sturgeon （20 mOsm· kg^ -1 ）. Second, it had a significant（ P≤0. 05 or P≤0.01 ）influence on VCL, VSL, VAP and percentage of motile sperm in cryopreserved samples. Third, pH of activating medium had the most significant difference on MAD （ P≤ 0.01 ） among the parameters, and it was lower and MAD was lager. Fourth, the most appropriate pH was 7. 5 for Siberian sturgeon and 8. 5 for Chinese sturgeon, Amur sturgeon and paddlefish. And last, the velocities（ VCL, VSL, VAP） had significant increases （P ≤0.01 ）, when MgCl2 （5 mmol· L^-1 ） was added into activating medium. In conclusion, the appropriate activating mediums for the cryopreserved sperm of Siberian sturgeon, Chinese sturgeon and Amur sturgeon, paddlefish were 10 mmol ·L^-1 Tris-HCl ＋ 5 mmol ·L^-1 MgCl2, pH 7.5, 20 mmol ·L^-1 Tris-HCl ＋ 5 mmol ·L^-

关 键 词：鲟鱼 计算机辅助分析系统 超低温保存 精子活力 

分 类 号：S917[农业科学—水产科学]