摘要
研究冷冻保存前平衡温度对精子结构的影响,为提高猪精子冷冻效果提供参考。新鲜猪精液(Ⅰ组)稀释后于17℃过夜保存(Ⅱ组),然后于冷冻Ⅰ液中4℃平衡1.5h(Ⅲ组),再加入预冷的冷冻Ⅱ液于4℃平衡45min(Ⅳ组)。通过精子形态正常率及活力检测,吖啶橙(AO)染色检测精子DNA完整性,碘化丙啶(PI)染色检测精子头部质膜完整性,低渗肿胀试验(HOS)检测精子尾部质膜完整性,异硫氰荧光素标记的花生凝集素(FITC-PNA)染色检测精子顶体完整性,并用扫描电镜(SEM)观察精子结构,评价冷冻保存前平衡温度对猪精子结构的影响。结果表明,与新鲜精液相比,Ⅱ组精液的精子DNA完整性、质膜完整性、顶体完整性等精子结构检测指标所受影响无统计学意义(P>0.05);Ⅲ组和Ⅳ组的精子结构检测指标明显受到影响(P<0.05),但Ⅲ组和Ⅳ组间无显著差异(P>0.05)。扫描电镜检测结果表明,17℃过夜保存精子未见异常,头、颈、尾结构完整,顶体完整、光滑;Ⅲ组精子头部质膜及头颈结合部有轻微损伤;Ⅳ组精子尾部和头颈结合部有不同程度的断裂。冷冻前从17℃降低至4℃及4℃平衡对猪精子结构有一定程度的损伤。
To investigate the effect of equilibration temperature before freezing on boar sperm structure and to provide reference for optimize the cryopreservation protocol of boar semen,fresh semen （ I group） was diluted and then preserved at 17;C for overnight time （ II group）.Next,the semen was mixed with I freezing liquid at the ratio of their volume 1.1 and then kept for 1.5 hour at 4℃ （III group）.Finally,the semen was mixed with II freezing liquid and then kept for 45minutes at 4℃ （IV group）.In order to evaluate the effect of equilibration temperature before freezing on boar sperm structure,we examined semen samples by the normal proportion and activity assay of sperm morphology,the acridine orange（AO） staining,propidium iodide （PI） staining, hypoosmotic swelling test （HOS）, fluorescein isothiocyanate labeled peanut （FITC-PNA） staining and scanning electron microscopy （SEM） observation.Compare with fresh sperm,all the indexes of sperm from II group decrease but no statistical difference between them （P〉0.05）.All the indexes of sperm from III and IV groups decrease obviously （P〈0.05）.But there was no significant difference between III group and IV group （P〉0.05）.Under SEM,sperm from II group did not show abnormal and showed intact structure including head,neck and tail and intact and smooth acrosome.Sperm from HI group showed lightly damage in head member and junction between head and neck.Sperm from IV group showed different degree fracture in tail and junction between head and neck. In conclusion, temperature change can cause sperm damage when sperms undergo the temperature drop from 17℃ to 4℃ and 4℃ equilibration before freezing.
作者
刘莉
赵倩
邱树磊
郝福星
吴植
张斌
武彩红
郑筱峰
LIU Li ZHAO Qian QIU Shu-lei HAO Fu-xing WU Zhi ZHANG Bin WU Cai-hong ZHENG Xiao-feng(Veterinary College of Yangzhou University, Yangzhou , Jiangsu , 225009, China Jiangsu Animal Husbandry and Veterinary College, Taizhou ,Jiangsu ,225300 ,China Institute of Animal Health Supervision of Xinxiang , Xinxiang , Henan , 45300, China)
出处
《动物医学进展》
北大核心
2017年第6期38-42,共5页
Progress In Veterinary Medicine
基金
江苏省自然科学基金项目(BK2008589)
江苏省"333工程"[苏农办人2013(3)号]
江苏农牧科技职业学院"凤凰人才工程"[(2014)30号]
江苏省教育厅"青蓝工程"培养对象(苏教办师[2017]5号)
关键词
精液冷冻
平衡温度
精子结构
猪
sperm freezing
equilibration temperature
sperm structure
boar
