摘要
[目的]探索适用于犬性别控制的流式细胞仪分离精子的方法。[方法]采集昆明犬和德国牧羊犬的精液,采用不同浓度(6、8、10、12μl/ml)的Hoechst 33342对精液进行染色,探讨不同浓度的荧光染料Hoechst 33342及公犬个体对精子分离效果的影响。[结果]当Hoechst 33342浓度分别为6、8、10μl/ml时,X和Y精子的DNA差异不能被分辨成双峰,只有当Hoechst 33342为12μl/ml时,X/Y双峰能有效并稳定地分离。不同公犬的鲜精死精率与X/Y精子分离速度呈明显负相关性,而分离准确率则与鲜精死精率无明显相关性。[结论]当Hoechst 33342浓度为12μl/ml时,流式细胞仪能将警犬X、Y精子成功分离。选择适宜的种公犬对提高精子的分离效率和分离精液的品质都有重要意义。
[Objective] The research aimed to explore the sex-sorting method of canine spermatozoa by using flow cytometry.[Method]The semen samples were collected from Kunming dogs and German shepherd dogs and stained with different concentrations（ 6,8,10 and 12 μl / ml） of Hoechst 33342. The influences of different concentrations of Hoechst 33342 and different dog individuals on the sex-sorting efficiency were discussed.[Result] When the concentrations of Hoechst 33342 was 6,8,10 μl / ml,fresh canine sperm could not be effectively separated. When the concentration of Hoechst 33342 was 12 μl / ml,fresh canine sperm could be effectively and stably separated. The death rate of sperm in fresh semen of different male dogs showed an obvious negative correlation with the sex-sorting speed,but the sex-sorting accuracy had no obvious correlation with the death rate of sperm in fresh semen.[Conclusion]When the concentration of Hoechst 33342 was 12 μl / ml,the sex-sorting of canine spermatozoa could be successfully performed by using flow cytometry. It was of great significance for improving the sex-sorting efficiency of sperm and the quality of separated sperm to select suitable male dog.
出处
《安徽农业科学》
CAS
2015年第9期95-98,123,共5页
Journal of Anhui Agricultural Sciences
基金
公安部应用创新计划项目(2011YYCXKMJQ167)
