Wang Yihan, Yuan Xiang, Ali Malik Ahsan, Qin Ziyue, Zhang Yan, Zeng Changjun
Farm Animal Genetic Resources Exploration and Innovation Key Laboratory of Sichuan Province, College of Animal Science and Technology, Sichuan Agricultural University, Chengdu, China.
Department of Theriogenology, Riphah College of Veterinary Sciences, Lahore, Pakistan.
Front Cell Dev Biol. 2022 Jan 26;9:792994. doi: 10.3389/fcell.2021.792994. eCollection 2021.
Cryopreservation induces capacitation-like (cryo-capacitation) changes, similar to natural capacitation, and affects the fertility potential of post-thawed sperm. The molecular mechanism of sperm cryo-capacitation during cryopreservation remains unknown. PIWI-interacting RNAs (piRNAs) have been reported to be involved in cryo-capacitation of post-thawed sperm and regulation of sperm motility, capacitation, and chemotaxis. In this study, protein tyrosine phosphatase nonreceptor type 7 (PTPN7) was positively targeted by piR-121380 after a dual luciferase assay. The mRNA expression of PTPN7 and piR-121380 was significantly decreased ( < 0.01); however, PTPN7 protein was significantly increased ( < 0.01) in post-thawed boar sperm. Furthermore, E1RK1/2 phosphorylation was reduced during cryopreservation. Six hours after transfection with piR-121380 mimic and inhibitor, the phosphorylation of ERK2 was significantly increased and decreased ( < 0.01), respectively. Furthermore, the highest and lowest total sperm motility, forward motility, and capacitation rate were observed after piR-121380 mimic and inhibitor treatments, respectively. The concentration of intracellular calcium ([Ca2+]i) showed no significant difference after transfection with either piR-121380 mimic or inhibitor at 1, 3, and 6 h. In conclusion, we demonstrated that piR-121380 modulates ERK2 phosphorylation by targeting PTPN7, which induces sperm cryo-capacitation, and eventually affects the motility and fertility potential of post-thawed sperm.
冷冻保存会诱导类似获能（冷冻获能）的变化，类似于自然获能，并影响解冻后精子的生育能力。冷冻保存过程中精子冷冻获能的分子机制尚不清楚。据报道，PIWI相互作用RNA（piRNA）参与解冻后精子的冷冻获能以及精子运动、获能和趋化性的调节。在本研究中，经过双荧光素酶测定后，蛋白酪氨酸磷酸酶非受体7型（PTPN7）被piR-121380正向靶向。解冻后公猪精子中PTPN7和piR-121380的mRNA表达显著降低（<0.01）；然而，PTPN7蛋白显著增加（<0.01）。此外，冷冻保存期间E1RK1/2磷酸化降低。用piR-121380模拟物和抑制剂转染6小时后，ERK2的磷酸化分别显著增加和降低（<0.01）。此外，分别在piR-121380模拟物和抑制剂处理后观察到精子总活力、前向运动能力和获能率的最高值和最低值。在1、3和6小时用piR-121380模拟物或抑制剂转染后，细胞内钙浓度（[Ca2+]i）没有显著差异。总之，我们证明piR-121380通过靶向PTPN7调节ERK2磷酸化，从而诱导精子冷冻获能，并最终影响解冻后精子的运动能力和生育潜力。
