美洲黑石斑精子超低温保存研究  

An efficient methodology of sperm cryopreservation in Centropristis striata

作　　者：刘洪军[1] 官曙光[1] 刘清华[2] 李军[2] 于道德[1] 

机构地区：[1]山东省海水养殖研究所,山东青岛266002 [2]中国科学院海洋研究所,山东青岛266071

出　　处：《海洋科学》2013年第1期76-80,共5页Marine Sciences

基　　金：农业部948项目(2011-Z39);中国科学院海洋研究所知识创新工程领域前沿项目(Y02507101Q);国家自然科学基金资助项目(41076100;31072212)

摘　　要：作者采用2 mL的冷存管和程序降温仪高效超低温保存美洲黑石斑(Centropristis striata L.)精子。分析比较了4种不同浓度抗冻剂15%DMSO(二甲基亚砜)、15%EG(乙二醇)、15%PG(丙二醇)和10%Meth(甲醇),5种降温速率(10,15,20,25,30℃/min)和5种解冻温度(30,35,40,45,50℃)对美洲黑石斑精子超低温保存效果的影响。实验结果表明,采用Hanks’作为稀释液,15%DMSO、15%EG、15%PG作为抗冻剂,30℃/min的降温速率对精液进行超低温保存35℃水浴解冻,冻精激活后获得理想的运动率(>60%)。通过对抗冻剂、降温速率、解冻温度的筛选,作者建立了美洲黑石斑精液高效超低温保存的方法,并对冻精进行长期保存。美洲黑石斑精子超低温保存方法的建立对于海水鱼类精子库的建立以及种质资源的保存和生物多样性的保护具有重要意义。In the present study, Centropristis striata sperm was efficiently cryopreserved using a programmable freezer. The motility of both fresh and post-thaw sperm was investigated in order to optimize the sperm cryopreservation protocols for C. striata. Four cryoprotectants （15% dimethyl sulfoxide, 15% ethylene glycol, 15% pro- pylene glycol and 10% methanol）, five cooling rates （10, 15, 20, 25 and 30℃/min） as well as five thawing ternperatures （30, 35, 40, 45 and 50 ℃） were designed and tested in the sperm cryopreservation, and their effects on post-thaw sperm motility were studied. Optimal post-thaw motility （〉60%） were achieved when supplemented with 15% DMSO, 15% PG and 15% EG with 30℃/min cooling rate, and 35% water usmg bath. Hanks＇s An eftcient cryopreservation method for C. striata sperm was established by analyzing the effect of cryoprotectants, Cooling rates and thaw temperatures. This method is advantageous not only to establish laboratory experiment but also to preserve genetic resources for routine aquaculture hatchery operation

关 键 词：美洲黑石斑(Centropristis striata L ) 精子低温保存 抗冻剂 精子活力 

分 类 号：Q331[生物学—遗传学]