鲈鱼胚胎的玻璃化冷冻保存  

Cryopreservation of the sea perch (Lateolabrax japonicus) embryos by vitrification

作　　者：田永胜[1] 陈松林[2] 严安生[1] 季相山[2] 于过才[2] 

机构地区：[1]华中农业大学水产学院,武汉430070 [2]中国水产科学研究院黄海水产研究所

出　　处：《动物学报》2003年第6期843-850,共8页ACTA ZOOLOGICA SINICA

基　　金：国家"十五" 863项目 (No :2 0 0 1AA62 110 0 );农业部海洋渔业资源可持续利用重点开放实验室开放课题~~

摘　　要：本文对鲈鱼 (Lateolabraxjaponicus)胚胎进行了玻璃化冷冻保存研究 ,筛选出了浓度较低、玻璃化程度较稳定的 5种玻璃化液 ,冷冻时形成玻璃化的概率在 4 8 1%～ 10 0 % ,在 35～ 4 3℃的水浴中解冻时保持玻璃化的概率在 4 4 4 %～ 6 3 0 % ;玻璃化液VSD2在解冻时保持玻璃化的概率最高。对鲈鱼神经胚、 2 0对肌节胚、尾芽胚、心跳胚、出膜前胚在玻璃化液VSD2中的适应能力及适合于玻璃化冷冻的胚胎时期进行了比较 ,结果显示 :不同时期胚胎对玻璃化液的耐受能力不同 ,鲈鱼神经胚耐受能力最低 ,心跳胚耐受能力最强 ,出膜前期胚次之 ,心跳胚和出膜前胚适合于进行玻璃化冷冻。对 0 5mol/L蔗糖的洗脱时间进行了选择 ,结果显示 ,洗脱10～ 2 0min效果较好。利用玻璃化程度较好的VSD2对鲈鱼不同时期胚胎进行超低温 ( - 196℃ )冷冻 ,获得了2 1%～ 2 7 9%的透明胚。将鲈鱼心跳胚冷冻解冻后获 2粒复活胚 ,培养至出膜期 ,成活 4 2～ 5 0h ;出膜前期胚在冷冻解冻后有 1粒胚复活 。To improve cryopreservtion of sea perch ( Lateolabrax japonicus ) embryos, we selected five vitrification solutions that could vitrify consistently. The cryoprotectants concentration in the five vitrification solutions were low to reduce toxicity. The vitrification probability of the five solutions was 48 1%-100% during freezing and 44 4%-63 0% during thawing at 35-43℃ water bath, respectively. VSD2 was more stable during thawing than the other four vitrification solutions and was used to cryopreserve sea perch embryos in the experiment. The durable time of sea perch neurula stage, 20 pair muscle stage, tail bud stage, heart beating stage and prehatching stage in VSD2 was compared and the most suitable stage to cryopreserve by vitrification was selected. We found that the neurula stage sustained the least time in VSD2 and the heart beating stage was the most suitable stage to cryopreserve by vitrification. The pre hatching stage was also feasible to cryopreserve by vitrification. The effect of different elution times in 0 5 mol/L sucrose was compared and elution for 10-20min in 0 5 mol/L sucrose was found to be efficient. We used VSD2 as vitrification solution to cryopreserve(-196℃) sea perch embryos of different stages and got 2 1%-27 9% transparent embryos after thawing. Furthermore, two frozen thawed embryos at heart beating survived 42-50 h after thawing. Another frozen thawed embryo at pre hatching stage hatched.

关 键 词：鲈鱼 胚胎 玻璃化 冷冻 

分 类 号：S961[农业科学—水产养殖]