Liu Tengfei, Han Yawen, Zhou Ting, Zhang Ruihang, Chen Hong, Chen Shulin, Zhao Huiying
College of Veterinary Medicine, Northwest A & F University, Yangling 712100, Shaanxi Province, China.
Aging (Albany NY). 2019 Sep 23;11(18):7880-7898. doi: 10.18632/aging.102295.
Liquid storage of spermatozoa is important for artificial insemination and herd genetic breeding. However, the extended time of storage inducing the rapid decline in spermatozoa quality limits the development of this technology. The molecular mechanisms underlying liquid storage of spermatozoa remain largely unexplored. In this study, the effects of liquid storage on functional quality of spermatozoa were assessed in goat (). The time-dependent decline in spermatozoa motility showed a strong correlation with the significant increase in apoptosis. Moreover, apoptosis-related ultrastructural changes were observed, especially the defects in mitochondria. A significant decrease in mitochondrial membrane potential and changes in the expression of mitochondrial apoptosis-related proteins indicated mitochondrial dysfunction and mitochondrial apoptotic pathway activation. Notably, the abnormally high level of reactive oxygen species (ROS) caused by liquid storage resulted in oxidative damage to mitochondria and accelerated mitochondria-dependent apoptosis, as demonstrated by the addition of ROS scavenger N-acetylcysteine. Furthermore, critical differentially expressed proteins involved in mitochondria-dependent apoptosis and antioxidant defense were identified and profiled by quantitative proteomic analysis, facilitating the understanding of molecular regulation of ROS-induced mitochondria-dependent apoptosis. These outcomes provide insights into the mechanisms underlying liquid storage of goat spermatozoa and enhance the progress of semen storage technology.
精子的液体保存对于人工授精和畜群遗传育种很重要。然而，延长保存时间会导致精子质量迅速下降，这限制了该技术的发展。精子液体保存的分子机制在很大程度上仍未被探索。在本研究中，评估了液体保存对山羊精子功能质量的影响。精子活力随时间的下降与凋亡的显著增加密切相关。此外，观察到与凋亡相关的超微结构变化，尤其是线粒体的缺陷。线粒体膜电位的显著降低以及线粒体凋亡相关蛋白表达的变化表明线粒体功能障碍和线粒体凋亡途径的激活。值得注意的是，液体保存导致的活性氧（ROS）异常高水平对线粒体造成氧化损伤并加速线粒体依赖性凋亡，添加ROS清除剂N-乙酰半胱氨酸证明了这一点。此外，通过定量蛋白质组分析鉴定并分析了参与线粒体依赖性凋亡和抗氧化防御的关键差异表达蛋白，有助于理解ROS诱导的线粒体依赖性凋亡的分子调控。这些结果为山羊精子液体保存的机制提供了见解，并促进了精液保存技术的进步。
