Grieblová Adéla, Pintus Eliana, Ros-Santaella José Luis
Department of Veterinary Sciences, Faculty of Agrobiology, Food and Natural Resources, Czech University of Life Sciences Prague, Kamýcká 129, 16500, Prague 6-Suchdol, Czech Republic.
Department of Veterinary Sciences, Faculty of Agrobiology, Food and Natural Resources, Czech University of Life Sciences Prague, Kamýcká 129, 16500, Prague 6-Suchdol, Czech Republic.
Anim Reprod Sci. 2017 Sep;184:218-227. doi: 10.1016/j.anireprosci.2017.07.020. Epub 2017 Jul 29.
An intact and functional sperm plasmalemma has a major role in sperm motility and fertilizing capacity. Several techniques have been developed to evaluate the integrity of the sperm plasma membrane, but there are still some inconsistencies concerning the methods that are more closely associated with sperm function. In this study, the aim was to: i) evaluate the integrity of the boar sperm plasmalemma during 72h of semen storage at 17°C using four techniques: eosin/nigrosin (E/N), propidium iodide/carboxyfluorescein diacetate (PI/CFDA), hypo-osmotic swelling test (HOST), and combined HOST with eosin staining (HOST/E), ii) assess the correlations and the limits of consistency among these techniques, iii) and estimate the relationships with the acrosomal status and sperm kinetics. Results indicate that the integrity of the sperm plasmalemma decreases during 72h of storage, although significant differences were found only using the HOST and HOST/E techniques. Moreover, use of E/N and PI/CFDA results in greater values relative to the undamaged sperm membrane than use of HOST and HOST/E at any incubation time. Overall, results using all techniques were consistent and correlate except for findings with PI/CFDA and HOST, which was slightly below 95%. Moreover, values using the techniques for the evaluation of the integrity of the sperm head and tail membranes are positively associated with the acrosomal status and different kinetic variables with the tail integrity being related to rapid linear trajectories and the head integrity to rapid curvilinear trajectories. The results of this study provide new insights into the relevance of evaluating the boar sperm plasmalemma in the routine spermiogram.
完整且功能正常的精子质膜在精子活力和受精能力方面发挥着重要作用。已经开发了多种技术来评估精子质膜的完整性，但在与精子功能更密切相关的方法方面仍存在一些不一致之处。在本研究中，目的是：i）使用四种技术评估17°C精液储存72小时期间公猪精子质膜的完整性：伊红/黑色素（E/N）、碘化丙啶/羧基荧光素二乙酸酯（PI/CFDA）、低渗肿胀试验（HOST）以及HOST与伊红染色相结合（HOST/E）；ii）评估这些技术之间的相关性和一致性限度；iii）估计与顶体状态和精子动力学的关系。结果表明，在储存72小时期间精子质膜的完整性下降，尽管仅使用HOST和HOST/E技术时发现了显著差异。此外，在任何孵育时间，与未受损精子膜相比，使用E/N和PI/CFDA得到的值更高。总体而言，除了PI/CFDA和HOST的结果略低于95%外，使用所有技术得到的结果都是一致且相关的。此外，用于评估精子头部和尾部膜完整性的技术的值与顶体状态和不同的动力学变量呈正相关，尾部完整性与快速直线轨迹相关，头部完整性与快速曲线轨迹相关。本研究结果为在常规精子检查中评估公猪精子质膜的相关性提供了新的见解。
