三疣梭子蟹精子保存研究  

Preservation of spermatozoa of blue crab Portunus trituberculatus

作　　者：周帅[1,2] 朱冬发[1] 王春琳[1] 薛良义[1] 

机构地区：[1]宁波大学生命科学与生物工程学院 [2]杭州二中树兰实验学校,浙江杭州311100

出　　处：《海洋科学》2007年第7期37-42,共6页Marine Sciences

基　　金：国家自然科学基金资助项目(40576068);浙江省自然科学基金资助项目(Y305339)

摘　　要：于2004年5月到2004年7月对三疣梭子蟹精子进行了常温、低温和超低温保存研究。利用离子载体A23187人工诱导顶体反应以判断精子活力。常温和低温保存实验的结果显示,4℃下无钙人工海水(calcium-free artificial sea water,Ca2+FASW)保存236h的精子有25%以上的成活率。在4℃,25℃两种温度与Ca2+FASW、天然海水(natural sea water,NSW)两种基础液构成的4种组合中,最优组为4℃、Ca2+FASW,最差组为25℃、NSW。将精子浓度与Ca2+FASW对照组及诱导组比较,结果显示,25℃和4℃下,精子浓度与对照组顶体反应率相关性不显著(P=0.068>0.05,P=0.265>0.05),与诱导组顶体反应率极显著相关(P<0.01)。以二甲亚砜(Dimethyl sulfoxide,DMSO)和甘油(Glvcerol,G)作为抗冻剂进行超低温冷冻实验,72h后解冻,包括对照组在内的所有组均有98%以上的精子不经诱导就发生顶体反应。Normal（25 ℃）,low temperature（4 ℃） preservation and cryopreservation（- 196 ℃） of spermatozoa of blue crab Portunus trituberculatus were studied during May 2004 to July 2004. The viability assay of spermatozoa was carried out by means of the induction acrosome reaction method. The results of normal low temperature preservation showed that over 25% of acrosome reaction rate was achieved by treating the spermatozoa with ionophore A23187 when the sample was extended by calcium-free artificial sea water（Ca^2＋ FASW） at 4 ℃ for 236 hours. The best preservation choice was to adopt the group of 4 ℃, Ca^2＋FASW in the four group of 4 ℃ Ca^2＋ FASW, 25 ℃ Ca^2＋ FASW, 4 ℃ natural sea water（NSW ） , 25 ℃ NSW and the worst one was to adopt the group of 25 ℃ NSW. Rates of acrosome reaction and concentration of sperm in Ca^2＋ FASW at 25 ℃,4 ℃ were compared and analyzed. It was conclued that there were not significant correlation between the rates of acrosome reaction in control and concentration of sperm,but there were very significant positive correlation between the rates of acrosome reaction induced artificially by treatment with ionophore A23187 and concentration of sperm（P〈0.01）.After 72 hours,the cryopreserved spermatozoa sample extended by the DMSO or glvcerol had over 98% rate of acrosome reaction including the control without any induction.

关 键 词：三疣梭子蟹(Portunus trituberculatus) 精子 保存 

分 类 号：Q959[生物学—动物学]