作　　者： ; ; ;

机构地区： 广东海洋大学水产学院

出　　处： 《广东海洋大学学报》 2011年第6期86-90,共5页

摘　　要： 采用罗丹明123(Rh123)和碘化丙啶(PI)两种荧光染料对栉江珧(Atrina pectinata)精子线粒体活性进行检测,比较不同染液浓度和染色时间的检测效果,并用该方法检测和比较超低温冷冻保存对线粒体活性的影响。结果显示:Rh123和PI染色的最适染液浓度均为10μg/mL,染色时间控制在10 min之内;染色后具有线粒体活性的精子发出绿色荧光,死精子发出红色荧光,质膜受损但线粒体还未受损的精子发出红绿色荧光,表明Rh123和PI双荧光染色检测栉江珧精子线粒体活性是可行的;精液经冷冻后,线粒体活性为61.9%、质膜完整率56.8%,显著低于新鲜精液组(95.3%、93.5%),表明冷冻对精子结构会造成不同程度的损伤,Rh123和PI双荧光染色法可用来检测栉江珧精子质量。 Two fluorescent dyes,Rhodaminel23（Rh123） and Propidium iodide（PI）,were used to detect the mitochondrial activity of the spermatozoa of Atrina pectinata,and the testing effect of different staining time and dye concentration were also compared,and the mitochondrial activity of sperm after cryopreservation（frozen in liquid nitrogen for 24 h） was detected.The result showed that the most effective staining appeared when the concentration of Rhodaminel23 and Propidium iodide were 10 μg/mL,and the optimum reaction time was 10 min.The activity of mitochondria sperm emitted green fluorescence,and dead sperm emitted red fluorescence,and sperm with damaged plasmalemma and undamaged mitochondria sent out red-green fluorescence.Therefore,Rh123-PI double staining was feasible.Frozen sperm’s mitochondrial activity and plasma membrane integrity was 61.8%,56.8%,respectively,and was lower than fresh sperm（95.3%,93.5%）.The result indicated that the freezing-thawing process damaged sperm structure and Rh123-PI double staining could detect the quality of the spermatozoa of Atrina pectinata.

关 键 词： 栉江珧 精子 线粒体 罗丹明 碘化丙啶 荧光染色

领　　域： [生物学] [生物学]