动物医学进展Issue(9)：58-62,5.
单细胞凝胶电泳检测南美白对虾冷冻精子的 DNA 损伤
Detection of DNA Damage of Cryopreservation Spermatozoa in Litopenaeus vannamei Using Single Cell Gel Electrophoresis
摘要
Abstract
The DNA damage of cryopreservation spermatozoa in Litopenaeus vannamei was detected to as-sess the effect of cryopreservation,using a single cell gel electrophoresis assay (SCGE)method.The sperm were pretreated,electrophoresed in the alkaline electrophoresis solution,dyed with GelRed for 10 min,ob-served under fluorescence microscope,counted the proportion of nuclei with tails in about 50 nuclei at ran-dom,and then calculated the comet rate.Various parameters of DNA migration were analyzed and meas-ured by CASP analysis software,and the sperm nuclei were graded according to the length of comet tail and the fluorescence intensity.Compared with the DNA damage states of Litopenaeus vannamei frozen-thawed sperm and the fresh sperm,the result indicated that when the concentration of DMSO went up,the tail DNA,tail length,tail moment and Olive tail moment increased,and the comet rate and damage coeffi-cient increased too.The fresh sperm were basically no damage,the groups of 50 mL/L DMSO were slight-ly damaged,the groups of 100 mL/L and 150 mL/L DMSO were mainly slightly and midly damaged,the groups of 250 mL/L DMSO were more midly and heavily damaged,but totally damage was few in the sperm for this study.Our analysis suggests that the concentration of DMSO is the chief factor that causes the DNA damage in cryopreserved sperm nuclei.关键词
南美白对虾/精子/超低温冷冻保存/单细胞凝胶电泳/DNA 损伤Key words
Litopenaeus vannamei/sperm/cryopreservation/single cell gel electrophoresis(SCGE)/DNA damage分类
农业科技引用本文复制引用
杨春玲,赵永贞,彭敏,陈晓汉,李咏梅,陈秀荔..单细胞凝胶电泳检测南美白对虾冷冻精子的 DNA 损伤[J].动物医学进展,2014,(9):58-62,5.基金项目
广西科技攻关项目（桂科攻1222013-1）；广西水产遗传育种与健康养殖重点实验室开放课题项目 （）
