实验动物与比较医学 ›› 2015, Vol. 35 ›› Issue (6): 484-489.DOI: 10.3969/j.issn.1674-5817.2015.06.011

• 论著 • 上一篇    下一篇

拉布拉多犬X/Y 精子分选及性别控制冷冻精液制备实验研究

高一龙, 秦海斌, 温海, 强京宁, 贺星亮, 张汇东, 马大君, 朱骞, 李刚   

  1. 公安部南京警犬研究所, 警犬技术公安部重点实验室, 南京 210012
  • 收稿日期:2015-05-26 出版日期:2015-12-25 发布日期:2015-12-25
  • 作者简介:高一龙(1982-), 男, 硕士, 助理研究员, 研究方向: 主要从事警犬遗传育种、繁殖和行为遗传学研究。E-mail: gaoyilong726@163.com
  • 基金资助:
    公安部重点项目“性别控制技术在警犬培育中 的应用”(201201ZDYJ007)

Study on Cryopreservation of Flow-cytometrically Sorted X-or Y- chromo some Bearing Sperm on Labrador Retriever

GAO Yi-long, QIN Hai-bin, WEN Hai, QIANG Jing-ning, HE Xing-liang, ZHANG Hui-dong, MA Da-jun, ZHU Qian, LI Gang   

  1. Nanjing Policedog Institute of the Ministry of Public Security,Policedog Technology Key Laboratory of MPS, Nanjing 210012, China
  • Received:2015-05-26 Online:2015-12-25 Published:2015-12-25

摘要: 目的 探讨流式细胞分选技术在分离拉布拉多犬X、Y 精子方面的应用,并研究拉布拉多犬性别控制(性控)冷冻精液的制备方法。方法 人工按摩刺激法采集8只拉布拉多种公犬精液, 利用SX_MOFLO流式细胞仪展开X、Y 精子分离实验,并对分离后的X/Y精子冷冻、解冻后进行精液重分析和品质鉴定。 结果 流式细胞仪分选的X/Y精子纯度达到89%以上(89%~91%),X精子平均分离速度(4 260 个/s) 极显著高于Y 精子分离速度(3 930个/s)(P<0.01); 常规冷冻精液和X、Y 型冷冻精液性别比差异显著(P<0.05); 冻融后活率则无明显差异(P>0.05) ; 常规冷冻精液与X、Y 型冷冻精液解冻后1 h存活指数差异明显(P<0.05) ,分别为0.251±0.006、0.054±0.007 和0.048±0.006; 冻融后顶体完整率差异显著(P<0.05), 分别为77.00%±0.98%、84.66%±1.22%和84.01%±1.23%。 结论 用流式细胞分选技术可以获得高纯度的拉布拉多犬X/Y冷冻精液, 性别比和精液品质达到低剂量人工授精的要求。

关键词: 拉布拉多犬, 性别控制(性控), XY 精子, 流式细胞仪, 精液品质

Abstract: Objectives To investigate the application of flow cytometric technology in separating X or Y sperm from Labrador retriever, and to define a protocol for producing sex-sorted frozen semen. Methods The semen were collected from 8 Labrador retrievers by manual stimulation and transported in one hour, and then the semen were separated into X- and Y- chromosome by SX_MOFLO flow cytometric technology, a modified high-speed cell sorter, and finally frozen thawed. Sorted X- or Y-sperm and unsorted sperm as control were reanalyzed and quality assessed. Results The purities of flow sorted sex chromosomes in spermatozoa of three dogs ranged from 89% to 91%. The average separation velocity showed significantly differences between sorted X- sperm (4 260/s) and Y- sperm (3 930/s). The sex ratio showed significantly differences between frozen- thawed sorted X- or Y- chromosome bearing sperm (89.11%, 90.67%) and the control. The forward progressive motility was not significantly different between sorted sperm and the control. The survive index on sorted sperm was significantly lower (0.251±0.006\0.054±0.007, P<0.05) than the control (0.048±0.006). But the post thaw acrosome integrity on sorted sperm was significantly higher (84.66±1.22\84.01±1.23, P<0.05) than the control (77.00±0.98). Conclusion It was possible to obtain sorted frozen- thawed Labrador retriever sperm for AI by using flow cytometry techniques.

Key words: Labrador retriever, XY spermatozoa, Flow cytometry, Semen quality

中图分类号: