大黄鱼精液的高效超低温保存  

An efficient methodology of sperm cryopreservation of large yellow croaker(Pseudosciaena croce)

作　　者：肖志忠[1] 陈雄芳[2] 丁福红[1] 刘清华[1] 徐世宏[1] 李军[1] 

机构地区：[1]中国科学院海洋研究所海洋生物技术研究发展中心,山东青岛266071 [2]重庆邮电学院,重庆400065

出　　处：《海洋科学》2007年第4期1-4,72,共5页Marine Sciences

基　　金：国家863计划项目(2001AA621100;2003AA603510;2004AA603310)

摘　　要：采用2 mL的冷存管和程序降温仪高效超低温保存大黄鱼(Pseudosciaena croce)精液。分析比较了5种不同浓度抗冻剂(二甲基亚砜、乙二醇、丙二醇和甘油和甲醇)、3种降温速率(10,20和30℃/min)和2种解冻温度(28℃和43℃)对大黄鱼精液超低温保存效果的影响。实验结果表明,采用Hanks’作为稀释液,10%～20%甘油或10%DMSO作为抗冻剂,20℃/min的降温速率对精液进行超低温保存43℃水浴解冻,冻精激活后获得理想的运动率(76.6%±11.4%～80.0%±12.6%),而且对10%甘油保存的冻精进行人工授精实验获得了与新鲜精液相似的受精率和孵化率。In the present study, a great deal of yellow croaker （Pseudosciaena croce） sperm were efficiently cryopreserved in 2 mL cryovials by using a programmable freezer. The motility and fertility of both fresh and post-thaw sperm were investigated in order to optimize the spermatozoa cryopreservation protocols for red large yellow croaker. Five cryoprotectants with different concentrations, three cooling rates （10,20 and 30℃/min） from 0℃ to -180℃as well as two thawing temperatures （28℃ and 43℃） were designed and tested in the sperm cryopreservation, and their effects on post-thaw sperm motility were studied. Optimal post-thaw motility （76. 6± 11.4% -80. 0% ± 12. 6%） was achieved when using Hanks＂ supplemented with 10% to 20% glycerol or 10% dimethyl sulfoxide in combination with 20℃/min cooling rate, and thawing in 43℃ water bath. Furthermore, optimal fertilization rate of sperm frozen with 10% glycerol and hatching rate were obtained, which were similar to those of fresh sperm.

关 键 词：超低温保存 抗冻剂 大黄鱼(Pseudosciaena croce) 精液 运动率 受精率 

分 类 号：Q25[生物学—细胞生物学]