摘要
以葡萄牙牡蛎精子为对象,采用碘化丙啶(PI)/罗丹明123(Rh123)双染与流式细胞术(FCM),研究了不同盐度胁迫(10、15、20、25、30、35)对精子质量的影响,并对受胁迫精子进行授精实验。结果显示,随着盐度升高,精子的存活率、运动时间和活力先升高后降低;葡萄牙牡蛎精子的适宜盐度为20～35,活力较高的盐度范围为25～35;通过双染和FCM检测了精子质膜完整性和线粒体活性,发现盐度胁迫先对精子质膜造成损伤,后对线粒体活性造成伤害;低盐胁迫(10、15)对精子损伤严重,胁迫15 min时质膜受损比例高达67.26%±2.35%。人工授精结果显示,低盐胁迫下精子受精率和卵裂率显著降低,且卵裂阶段出现畸形及分裂停止等现象,表明精子质量不仅严重影响了受精过程,还对卵裂造成一定影响。本实验不仅探究了PI/Rh123双染和FCM检测牡蛎精子质膜完整性和线粒体活性的可行性,还结合显微观察全面评价了精子质量,为牡蛎精子质量的研究提供了理论基础。
In this experiment, Crassostrea angulata sperm was used as the research object to study the effect of salinity stress on sperm quality and fertilization rate using PI/Rh123 dual fluorescent staining and flow cytometry（FCM）. The results showed that sperm moving-time, survival rate and sperm activity increased first and then decreased with the increase of salinity. The suitable salinity range of C. angulata sperm was 20–35, and the salinity range was 25–35 with higher sperm activity. It was found that salinity stress caused damage to the sperm plasma membrane first and then damaged the mitochondrial activity by PI/Rh123 dual fluorescent staining and FCM. In addition, sperm were severely damaged under low salinity conditions（10, 15） and the damage ratio of sperm plasma membrane was 67.26%±2.35%, which was significantly different from the other groups. The artificial insemination experiment showed that fertilization rate and cleavage rate decreased significantly after low salinity stress（10, 15）, and deformity and split stagnation were observed in cleavage stage, which shows that sperm quality not only seriously affects the process of fertilization, but also has a certain effect on cleavage. This experiment explored the feasibility of PI/Rh123 dual fluorescent staining and FCM to detect the function of plasma membrane and mitochondrial activity in oyster sperm, but also evaluated the sperm quality comprehensively, which provided a theoretical basis for the research of oyster sperm quality.
作者
杜俊鹏
王昭萍
崔玉婷
李阳春
杨洋
李鹏飞
柳逸群
DU Junpeng1, WANG Zhaoping1, CUI Yuting1, LI Yangchun1, YANG Yang1, LI Pengfei2, LIU Yiqun1(1. Key Laboratory of Mariculture, Ministry of Education, Ocean University of China, Qingdao 266003, China; 2. Laizhou Changyu Fisheries Limited Company, Yantai 264003, China)
出处
《水产学报》
CAS
CSCD
北大核心
2018年第11期1737-1746,共10页
Journal of Fisheries of China
基金
国家自然科学基金(31172403)~~
