Influence of porcine spermadhesins on the susceptibility of boar spermatozoa to high dilution
Influence of porcine spermadhesins on the susceptibility of boar spermatozoa to high dilution
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DOI：
10.1095/biolreprod.103.016527
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发表时间：
2003-08-01
影响因子：
3.6
通讯作者：
Martinez, EA
中科院分区：
文献类型：
--
作者：
Centurion, F;Vazquez, JM;Martinez, EA
研究了在流式细胞术性别分选过程中精子暴露于高稀释度（300 000 个精子/ml）时，肝素结合和非肝素结合精子粘附素对公猪精子活力、运动性和线粒体活性的影响。用肝素结合精子粘附素孵育精子会导致功能性精子百分比出现时间和剂量依赖性下降。在 38°C 下与稀释在 PBS（1 mg/ml）中的肝素结合精子粘附素一起孵育的活精子百分比从 75%（0.5 小时）下降到 4%（5 小时），而在不含蛋白质的 PBS（对照）中孵育的活精子百分比从 85%（0.5 小时）下降到 19%（5 小时）。向 PBS 中添加非肝素结合的 PSP-I/PSP-II 精子粘附素会导致活细胞百分比呈浓度依赖性增加（孵育 5 小时后为 65%），在 1.5 mg/ml 时效果最大。孵育5小时后，肝素结合精子粘附素完全抑制精子活力和线粒体活性。在 1.5 mg/ml PSP-I/PSP-II 存在下孵育的精子的相同参数分别为 50% 和 58%，而对照精子表现出运动性和线粒体活性的百分比分别为 21% 和 26%。此外，当精子与 PSPI/PSP-II 和肝素结合精子粘附素的混合物一起孵育时，随着后者浓度的增加，活力、运动性和线粒体活性均下降。我们得出结论，PSP-I/PSP-II 和肝素结合精子粘附素对高度稀释的公猪精子的功能产生拮抗作用。 PSP-I/PSP-II 有助于在生理温度下维持精子的高活力、运动性和线粒体活性至少 5 小时，这一发现表明其有可能用作精子保存添加剂，特别是用于性别预选的高度稀释、流式分选的精子。
The effect of heparin-binding and non-heparin-binding spermadhesins on the viability, motility, and mitochondrial activity of boar spermatozoa at the high dilution (300 000 sperm/ml) to which sperm are exposed during the process of sex sorting by flow cytometry was investigated. Incubation of spermatozoa with heparin-binding spermadhesins caused a time- and dose-dependent decrease in the percentage of functional spermatozoa. The percentage of viable spermatozoa incubated at 38degreesC with heparin-binding spermadhesins diluted in PBS (1 mg/ml) dropped from 75% (0.5 h) to 4% (5 h), whereas the percentage of viable spermatozoa incubated in PBS without proteins (control) decreased from 85% (0.5 h) to 19% (5 h). Addition of non-heparin-binding PSP-I/PSP-II spermadhesin to the PBS resulted in a concentration-dependent increment of the percentage of viable cells (65% after 5-h incubation), with maximum effect at 1.5 mg/ml. The heparin-binding spermadhesins totally suppressed sperm motility and mitochondrial activity after 5 h of incubation. The same parameters of sperm incubated in the presence of 1.5 mg/ml of PSP-I/PSP-II were 50% and 58%, respectively, and the percentages of control sperm displaying motility and mitochondrial activity were 21% and 26%, respectively. Moreover, the viability, motility, and mitochondrial activity all decreased on incubation of spermatozoa with mixtures of PSPI/PSP-II and heparin-binding spermadhesins as the concentration of the latter increased. We conclude that PSP-I/PSP-II and the heparin-binding spermadhesins exert antagonistic effects on the functionality of highly diluted boar spermatozoa. The finding that PSP-I/PSP-II contributes to maintaining sperm with high viability, motility, and mitochondrial activity for at least 5 h at physiological temperature points to its potential use as an additive for sperm preservation, specifically of highly diluted, flow-sorted spermatozoa for sex preselection.