Ren Hongyan, Zhang Yandi, Bi Yanzhen, Wang Heng, Fang Guijie, Zhao Pengxiang
Hubei Key Laboratory of Animal Embryo Engineering and Molecular Breeding, Hubei Academy of Agricultural Sciences, Wuhan, Hubei Province, PR China.
College of Animal Science and Technology, Shandong Agricultural University, Taian, PR China.
Theriogenology. 2024 Apr 15;219:138-146. doi: 10.1016/j.theriogenology.2024.02.030. Epub 2024 Feb 28.
The quality of sperm significantly influences the reproductive efficiency of pig herds. High-quality sperm is necessary for efficient fertilization and to maximize the litter numbers in commercial pig farming. However, the understanding of genes regulating porcine sperm motility and viability is limited. In this study, we validated porcine sperm/Sertoli-specific promoters through the luciferase reporter system and identified vital genes for sperm quality via loss-of-function means. Further, the shRNAs driven by the ACE and SP-10 promoters were used to knockdown the SPAG6 and PPP1CC genes which were provisionally important for sperm quality. We assessed the effects of SPAG6 and PPP1CC knockdown on sperm motility by using the sperm quality analyzer and flow cytometry. The results showed that the ACE promoter is active in both porcine Sertoli cells and sperms, whereas the SP-10 promoter is operating exclusively in sperm cells. Targeted interference with SPAG6 and PPP1CC expression in sperm cells decreases the motility and increases apoptosis rates in porcine sperms. These findings not only offer new genetic tools for targeting male germ cells but also highlight the crucial roles of SPAG6 and PPP1CC in porcine sperm function.
精子质量显著影响猪群的繁殖效率。优质精子对于高效受精以及在商业养猪业中使产仔数最大化是必要的。然而，对调控猪精子活力和生存能力的基因的了解有限。在本研究中，我们通过荧光素酶报告系统验证了猪精子/支持细胞特异性启动子，并通过功能丧失手段鉴定了对精子质量至关重要的基因。此外，由ACE和SP-10启动子驱动的短发夹RNA（shRNAs）被用于敲低对精子质量暂时重要的SPAG6和PPP1CC基因。我们使用精子质量分析仪和流式细胞术评估了SPAG6和PPP1CC基因敲低对精子活力的影响。结果表明，ACE启动子在猪支持细胞和精子中均有活性，而SP-10启动子仅在精子细胞中起作用。对精子细胞中SPAG6和PPP1CC表达的靶向干扰降低了猪精子的活力并增加了凋亡率。这些发现不仅为靶向雄性生殖细胞提供了新的遗传工具，也突出了SPAG6和PPP1CC在猪精子功能中的关键作用。
