背景:体外获能在辅助生殖技术(ART)中对于胚胎产生至关重要。最近,精氨酸已被证明可增强哺乳动物精子的获能。然而,精氨酸的详细作用机制仍然难以捉摸。
目的:本研究研究了精氨酸诱导的获能和运动性增强对山羊精子RNA(spRNA)群体的影响。
方法:用精氨酸处理山羊精子长达6小时,并与未处理或PHE(青霉胺,亚牛磺酸,和肾上腺素)以不同的间隔(0、1、2、4和6小时)处理的精子。精子参数,包括生存能力,个体运动性,获能,顶体反应,和ROS生产,进行了评估。通过短读RNA测序(RNA-seq)分析spRNA群体。
结果:在精氨酸处理中,获能(73.21±4.22%)和顶体反应(18.35±0.56%)的精子的百分比最高,而PHE处理显示在孵育0至4小时内活动精子的百分比最高(79.82±4.31%)。与对照相比,RNA-seq分析在精氨酸处理的精子中鉴定了1,321个差异表达基因(DEG)。PGK2,RNASE10,ODF1和ROPN1L基因参与精子运动和ACR,参与获能过程的DKKL1,KCNJ11和PRND基因在精氨酸处理的精子中上调。DEGs调节精子获能相关的cAMP-PKA,PI3-Akt,钙,和MAPK信号通路。
结论:精氨酸诱导的获能和精子运动性增强与一些参与精子运动性和获能途径的基因上调有关。比较研究还表明,精氨酸可以代替PHE用于增强山羊精子的运动性和体外获能。
BACKGROUND: In vitro capacitation is essential in assisted reproductive technologies (ART) for embryo production. Recently, arginine has been proven to enhance capacitation in mammalian spermatozoa. However, the detailed mechanism of action of arginine remains elusive.
OBJECTIVE: This study investigated the effect of arginine-induced capacitation and motility enhancement on the spermatozoal RNA (spRNA) population in goats.
METHODS: Goat spermatozoa were treated with arginine for up to six hours and compared with non-treated or
PHE (penicillamine, hypotaurine, and epinephrine)-treated spermatozoa at different intervals (0, 1, 2, 4, and 6 hours). Sperm parameters, including viability, individual motility, capacitation, acrosome reaction, and ROS production, were evaluated. The spRNA population was analyzed by short-read RNA sequencing (RNA-seq).
RESULTS: The percentage of capacitated (73.21 ± 4.22%) and acrosome reacted (18.35 ± 0.56%) spermatozoa was highest in arginine treatment, while
PHE treatment showed the highest percentage (79.82 ± 4.31%) of motile spermatozoa from 0 to 4 hours of incubation. RNA-seq analysis identified 1,321 differentially expressed genes (DEGs) in arginine-treated spermatozoa compared to the control. The PGK2, RNASE10, ODF1, and ROPN1L genes involved in sperm motility and ACR, DKKL1, KCNJ11, and PRND genes involved in the capacitation process were upregulated in arginine-treated spermatozoa. The DEGs regulate sperm capacitation-related cAMP-PKA, PI3-Akt, calcium, and MAPK signaling pathways.
CONCLUSIONS: The arginine-induced capacitation and enhanced sperm motility were associated with the upregulation of several genes involved in sperm motility and capacitation pathways. The comparative study also suggests that arginine may be used in lieu of
PHE for motility enhancement and in vitro capacitation of goat spermatozoa.