The influence of antioxidant, cholesterol and seminal plasma on the in vitro quality of sorted and non-sorted ram spermatozoa
The influence of antioxidant, cholesterol and seminal plasma on the in vitro quality of sorted and non-sorted ram spermatozoa
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DOI：
10.1016/j.theriogenology.2006.07.008
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发表时间：
2007-01-15
期刊：
影响因子：
2.8
通讯作者：
O'Brien, J. K.
中科院分区：
文献类型：
--
作者：
de Graaf, S. P.;Evans, G.;O'Brien, J. K.
为了提高性别分选和冷冻保存后功能精子的数量，研究了添加剂对体外精子特征的影响：（i）过氧化氢酶（分选前）;（ii）胆固醇负载环糊精（CLC;分选前）;和（iii）精浆（解冻后）。对于所有实验，在冷冻保存、解冻和孵育6 h之前，使用高速流式细胞仪处理精子（3只雄性，n = 3次射精/雄性）。过氧化氢酶对性别分选的公羊精子解冻后的运动特性（如通过CASA测量的）没有影响（P > 0.05），但是分选前添加CLC降低了解冻后孵育0 h（运动性）、3 h（运动性、平均路径速度、活力和顶体完整性）和6 h（运动性、平均路径速度和曲线速度、直线性、线性、活力和顶体完整性）后的精子质量（P < 0.05）。精浆对性别分选精子和未分选精子的影响不同（P < 0.001）。解冻后补充增加水平的精浆导致性别分选的冷冻-解冻精子的所有运动特性下降（P < 0.05）;相反，未分选的冷冻-解冻精子在运动性、活力、顶体完整性和线粒体呼吸方面表现出改善（P < 0.05）。总之，将过氧化氢酶、CLC和精浆掺入分选方案中未能改善解冻后精子质量，因此不能提高公羊精子的性别分选效率。补充精浆对公羊精子体外特征的矛盾作用进一步证明了流式细胞术性别分选产生了与未分选精子相比具有不同功能的选定细胞群。(c)2006年爱思唯尔公司All rights reserved.
In an effort to improve the number of functional spermatozoa following sex-sorting and cryopreservation, the effects on in vitro sperm characteristics of the additives: (i) catalase (pre-sorting); (ii) cholesterol-loaded cyclodextrins (CLCs; pre-sorting); and (iii) seminal plasma (post-thawing) were investigated. For all experiments, spermatozoa (three males, n = 3 ejaculates/male) were processed using a high speed flow cytometer before cryopreservation, thawing and incubation for 6 h. Catalase had no effect (P > 0.05) on post-thaw motility characteristics (as measured by CASA) of sex-sorted ram spermatozoa, but pre-sort addition of CLCs reduced (P < 0.05) sperm quality after post-thaw incubation for 0 h (motility),:3 h (motility, average path velocity, viability and acrosome integrity) and 6 It (motility, average path and curvilinear velocity, straightness, linearity, viability and acrosome integrity). Seminal plasma had a differential effect (P < 0.001) on sex-sorted and non-sorted spermatozoa. Post-thaw supplementation of increasing levels of seminal plasma caused all motility characteristics of sex-sorted, frozen-thawed spermatozoa to decline (P < 0.05); conversely, non-sorted, frozen-thawed spermatozoa exhibited improvements (P < 0.05) in motility, viability, acrosome integrity and mitochondrial respiration. In summary, incorporation of catalase, CLCs and seminal plasma into the sorting protocol failed to improve post-thaw sperm quality and, consequently efficiency of sex-sorting of ram spermatozoa. The paradoxical effect of seminal plasma supplementation on the in vitro characteristics of ram spermatozoa provides further evidence that sex-sorting by flow cytometry produces a selected population of cells with different functions compared with non-sorted spermatozoa. (c) 2006 Elsevier Inc. All rights reserved.