猪精子质量的流式细胞检测分析

doi:10.16431/j.cnki.1671-7236.2016.09.041

Abstract

Abstract:

The flow cytometry was used in this trail to determine the pig sperm quality,in order to ensure the sperm had complete functions during the fertilization process.Four stain methods of SYBR-14/PI (Propidium Iodide),YO-PRO-1/PI,PNA/PI,Mito tracker/YO-PRO-1 were used to detect the plasma membrane integrity,apoptosis,top body ntegrity and mitochondrial function of Rongchang pig sperm.SYBR-14⁺/PI^- expressed live sperm with complete plasma membrane,YO-PRO-1^-/PI^- expressed live sperm,PNA^-/PI^- expressed live sperm had no top body reaction,and Mito Tracker⁺/YO-PRO-1^-expressed live sperm with high mitochondrial activity.The functional integrity of the tested sperm was evaluated according to the above comprehensive results.Through the study of Rongchang pig sperm quality detection by flow cytometry,it suggested that the method could provide theoretical basis for clinical diagnosis,and was very important for swine infertility treatments.

Key words: Rongchang pig; sperm; flow cytometry; detection

CLC Number:

S814.8

WU Meng, DING Yu-chun, LIU Zuo-hua, LIU Wen, LUO Lin, GE Liang-peng. Detection and Analysis of the Pig Sperm Quality by Flow Cytometry[J]. , 2016, 43(9): 2502-2507.

References

[1] 包宋权,梁世忠,韦明宇,等.聚维酮碘对常温保存猪精子质量的影响[J].中国兽医学报,2011,31(1):118-124.
[2] 潘红梅,张亮,蓝静,等.不同双糖组合对猪精子冷冻保存效果的影响[J].中国农学通报,2015,2:35-39.
[3] 乔利敏.动物精液质量检测方法研究进展[J].中国畜牧兽医,2013,40(7):153-157.
[4] 吴江.奶山羊精原细胞中miRNA表达谱和miR-204-Sirt1调控轴功能的研究[D].杨凌:西北农林科技大学,2014.
[5] 曾有权,陆阳清,杨小淦,等.使用流式细胞仪分离精子进行仔猪性别控制的研究[J].畜牧兽医学报,2012,43(7):1163-1169.
[6] Garner D L,Evans K M,Seidel G E.Sex-sorting sperm using flow cytometry/cell sorting[J].Spermatogenesis:Methods and Protocols,2013,927:279-295.
[7] 王昕,戴建军,吴彩凤,等.猪冷冻精子的细胞凋亡及凋亡途径初步研究[J].畜牧兽医学报,2015,46(8):1341-1347.
[8] Broekhuijse M,Šoštari? E,Feitsma H,et al.Relationship of flow cytometric sperm integrity assessments with boar fertility performance under optimized field conditions[J].Journal of Animal Science,2012,90(12):4327-4336.
[9] Zhou X,Xia X Y,Cao Q,et al.Evaluation of sperm plasma membrane integrity by SYBR-14/PI dual fluorescent staining and flow cytometry[J].National Journal of Andrology,2010,16(7):589-593.
[10] 叶玲玲,刘红,李世崇,等.用YO-PRO-1和PI联合染色定量检测细胞凋亡[J].中国生物工程杂志,2005,25(10):50-53.
[11] Robles V,Martínez-Pastor F.Flow cytometric methods for sperm assessment[J].Spermatogenesis:Methods and Protocols,2013,927:175-186.
[12] Martínez-Pastor F,Mata-Campuzano M,álvarez-Rodríguez M,et al.Probes and techniques for sperm evaluation by flow cytometry[J].Reproduction in Domestic Animals,2010,45(S2):67-78.
[13] 李新红,高超,陈翔宇,等.利用流式细胞仪评价不同防冻液对猪冻融精子成活率及凋亡状况的影响[J].上海交通大学学报:农业科学版,2007,25(5):468-473.
[14] Li W W,Li N,Wu Q Y,et al.Cigarette smoking affects sperm plasma membrane integrity[J].National Journal of Andrology,2012,18(12):1093-1096.
[15] 任康,史文清,路永强,等.不同保存条件对猪精子质膜,顶体及线粒体损伤的研究[J].猪业科学,2014,31(4):102-105.
[16] 王美姣.精子线粒体膜蛋Prohibitin和UPS通路相关蛋白的表达及与男性不育的相关性研究[D].上海:复旦大学,2012.
[17] Petrunkina A M,Waberski D,Günzel-Apel A R,et al.Determinants of sperm quality and fertility in domestic species[J]. Reproduction,2007,134(1):3-17.
[18] Lo K,Brinkman R R,Gottardo R.Automated gating of flow cytometry data via robust model-based clustering[J].Cytometry Part A,2008,73(4):321-332.
[19] Savi ? R,Petrovi? M,Radojkovi? D,et al.The effect of breed,boar and season on some properties of sperm[J].Biotechnology in Animal Husbandry,2013,29(2):299-310.
[20] 戴波,张明军.影响种公猪精液品质的因素及改进措施[J].湖南畜牧兽医,2015,3:24-26.

Detection and Analysis of the Pig Sperm Quality by Flow Cytometry

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