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半滑舌鳎精子冷冻保存
田永胜1, 陈松林1, 季相山1, 孙礼娟1, 翟介明1
农业部海洋渔业资源可持续利用重点开放实验室 中国水产科学研究院黄海水产研究所,青岛 266071
摘要:
半滑舌鳎精子冷冻保存对于人工繁殖育苗、杂交育种、雌核发育及其性别控制研究具有重要的意义,为此,本文对半滑舌鳎精子冷冻保存方法进行了研究。分别利用2.8 mol/L的二甲基亚砜(DMSO)、甘油(Gly)和1,2-丙二醇(PG)冷冻保存该鱼精子。结果显示,DMSO冷冻保存精子的活力较高。利用MPRS+2.8 mol/L DMSO以1∶05、1∶1、1∶1.5和1∶2的比例稀释并冷冻精子,1∶1比例在冻前能够抑制精子的运动,冻后活力可达82.50±3.54%,显著高于其他稀释比例(P<0.05)。分别利用冷冻保存液A(MPRS+2.8 mol/L DMSO)和B(TS-2+2.8 mol/L DMSO)稀释平衡精子,精子在A中的冻前快速运动时间、寿命分别为37.75±6.45 s和145.00±78.98 s,与鲜精无显著差异(P>0.05)。利用以上两种冷冻稀释液冷冻保存精子,精子在A液中的冻后活力和寿命分别可达53.50±6.69%和98.00±13.51 s,冷冻效果优于B液(P<0.05)。冷冻后精子的受精率和孵化率分别为55.00±5.00%和35.00±13.23%,受精率与鲜精无显著性差异(P>0.05),因此认为MPRS+2.8 mol/L DMSO可用于半滑舌鳎精子的冷冻保存。
关键词:  半滑舌鳎  精子  冷冻保存
DOI:
分类号:
基金项目:国家自然科学基金项目(30570259)、“国家”863项目(2006AA10A403)、农业科技成果转化资金项目(2007GB2C600174)和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(实开2005-03)共同资助
Cryopreservation of tongue sole Cynoglossus semilaevis sperm
Abstract:
Cryopreservation of tongue sole Cynoglossus semilaevis sperm offers benefits for artificial propagation, hybridization, gynogenesis and sex control.In order to develop a protocol to freeze tongue sole sperm,the effects of cryoprotectants, extenders and dilution ratio on motility,fertilization rate and hatching rate of frozen-thawed sperm were evaluated. Dimethylsulfoxide(DMSO)provided higher protection than glycerol and 1,2 propylene glycol during freezing and thawing.Motion time and life of sperm in A(MPRS +28 mol/L DMSO)and B(TS-2+2.8 mol/L DMSO)before freezing were 37.75±6.45 sec and 145.00±78.98 sec,which was not different from that of fresh sperm(P>0.05).The motility and life of frozen thawed sperm in A were 5350±669% and 98.00±13.51 sec,which was significantly higher than that of frozen thawed sperm in B.When sperm were diluted with A at a ratio of 1∶05,1∶1,1∶15 and 1∶2, the highest motility(82.5±3.54%, 1∶1 dilution ratio)of frozen-thawed sperm was obtained and which was significantly higher than that of other dilution ratio treatment.According to the results above, tongue sole sperm were cryopreserved successfully with MPRS+2.8 mol/L DMSO at a dilution ratio of 1∶1.As a result,the fertilization rate(55.0±5.0%) and hatching rate(35±13.23%) of frozen-thawed sperm were obtained,which was not significantly different from that of fresh sperm(P>0.05).
Key words:  Cynoglossus semilaevis  Semen  Cryopreservation