条纹锯精液超低温冷冻保存研究
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中国海洋大学 水产学院,中国科学院 海洋研究所,山东省 海水养殖研究所,中国海洋大学 水产学院,中国海洋大学 水产学院,中国科学院 海洋研究所

基金项目:

国家自然科学基金(31072212,41076100);国家“八六三”高技术研究发展计划(2012AA10A402);水产种质资源平台运行服务项目


Cryopreservation for spermatozoa of Centropristis striata
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Fisheries College,Ocean University of China,Institute of Oceanology,Chinese Academy of Sciences,Qingdao,Mariculture Institute of Shandong Province,Fisheries College,Ocean University of China,Fisheries College,Ocean University of China,Institute of Oceanology,Chinese Academy of Sciences,Qingdao

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    摘要:

    为建立条纹锯精液超低温冷冻保存方法,实验采用计算机辅助精子分析系统(CASA)分析了采用6种抗冻保护剂(GLY[甘油]、DMSO[二甲基亚砜]、PG[丙二醇]、EG[乙二醇]、METH[甲醇]、DMA[二甲基乙酰胺])在4种浓度(5%、10%、15%、20%,v/v)下对条纹锯精液的冷冻保存效果。结果发现,以HBSS为稀释液,采用程序降温仪分步降温冷冻保存条纹锯精液,37℃水浴解冻后的精子中,15% PG 作为抗冻保护剂的精子运动率最高,达到(93.1±0.9)%,与鲜精差异不显著(P>0.05),15% PG 作为抗冻保护剂的精子水浴解冻后精子的运动速度最高,平均直线速度、平均曲线速度、平均路径速度分别达到了(88.3±0.3)μm/s、(76.2±0.5) μm/s、(86.7±0.7) μm/s,与鲜精差异不显著(P>0.05)。在不同种类及不同浓度抗冻保护剂保护下,15% PG 作为抗冻保护剂的精子解冻后 1 min内运动率变化与鲜精差异不显著(P>0.05)。研究表明,15% PG为条纹锯最佳抗冻保护剂,可用于条纹锯精液的超低温冷冻保存。

    Abstract:

    In order to establish Centropristis striata sperm cryopreservation method, this paper used a computer-assisted sperm motion analysis system(CASA),and selected six cryoprotectants,such as glycerin(GLY), dimethyl sulfoxide(DMSO), propylene glycol(PG), ethylene glycol(EG), dimethyl sulfoxide(DMA), methanol(METH) at four concentrations (5%, 10%, 15% and 20%, V/V) in HBSS to find the influence of C. striata sperm cryopreservation. Results showed that the best cryopreservation protocol for C. striata was 15% PG as antifreeze protective agent in HBSS with step cooling scheme. After 37°C water bath thawed, motility of the sperm reached (93.1±0.9)%, which was higher than other combination groups and showed no significant difference with fresh sperm (P>0.05).The group of 15% PG performed best in average linear velocity, average velocity, and average path velocity at (88.3±0.3) μm/s, (76.2±0.5) μm/s, and (86.7±0.7) μm/s, which showed no significant difference with fresh sperm (P>0.05). In different cryoprotectants with different concentration, the motility of 15% PG within 1 min had no significant difference with fresh semen. Therefore, 15% PG was the best cryoprotectant for C. striata and could be applied in practice.

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韩龙江,刘清华,官曙光,纪利芹,温海深,李军.条纹锯精液超低温冷冻保存研究[J].水产学报,2014,38(10):1714~1721

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  • 收稿日期:2014-06-30
  • 最后修改日期:2014-08-01
  • 录用日期:2014-10-02
  • 在线发布日期: 2014-10-17
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