Feng Tianhao, Zhou Shushu, Shi Xiaodan, Zhang Xin, Zhang Jintao, Zhao Shuqin, Yang Xiaoyu, Meng Xuhui, Liu Mingxi
State Key Laboratory of Reproductive Medicine, The Fourth Affiliated Hospital of Nanjing Medical University, Nanjing Medical University, Nanjing, China.
State Key Laboratory of Reproductive Medicine, Department of Histology and Embryology, School of Basic Medical Sciences, Nanjing Medical University, Nanjing, China.
Transl Androl Urol. 2021 May;10(5):1988-1999. doi: 10.21037/tau-21-18.
Eukaryotic elongation factor-2 kinase () is a protein kinase associated with the calmodulin-induced signaling pathway and an atypical alpha-kinase family member. -mediated phosphorylation of eukaryotic translation elongation factor 2 () can inhibit the functionality of this protein, altering protein translation. Prior work suggests to be overexpressed in breast, pancreatic, brain, and lung cancers wherein it may control key processes associated with apoptosis, autophagy, and cell cycle progression. The functional importance of in the testes of male mice, however, has yet to be clarified.
A CRISPR/Cas9 approach was used to generate male -knockout mice, which were evaluated for phenotypic changes in epididymal or testicular tissues through histological and immunofluorescent staining assays. In addition, TUNEL staining was conducted to assess the apoptotic death of cells in the testis. Fertility, sperm counts, and sperm motility were further assessed.
Male -knockout mice were successfully generated, and exhibited normal fertility and development. No apparent differences were observed with respect to spermatogenesis, sperm counts, or germ cell apoptosis when comparing male and mice.
Male -knockout mice remained fertile and were free of any evident developmental or spermatogenic abnormalities, suggesting to be dispensable in the context of male fertility.
真核生物延伸因子2激酶（）是一种与钙调蛋白诱导的信号通路相关的蛋白激酶，属于非典型α激酶家族成员。介导的真核生物翻译延伸因子2（）磷酸化可抑制该蛋白的功能，从而改变蛋白质翻译。先前的研究表明，在乳腺癌、胰腺癌、脑癌和肺癌中过表达，可能在其中控制与细胞凋亡、自噬和细胞周期进程相关的关键过程。然而，在雄性小鼠睾丸中的功能重要性尚未阐明。
采用CRISPR/Cas9方法构建雄性基因敲除小鼠，通过组织学和免疫荧光染色分析评估附睾或睾丸组织的表型变化。此外，进行TUNEL染色以评估睾丸中细胞的凋亡死亡情况。进一步评估生育能力、精子数量和精子活力。
成功构建了雄性基因敲除小鼠，其生育能力和发育正常。比较雄性基因敲除小鼠和野生型小鼠时，在精子发生、精子数量或生殖细胞凋亡方面未观察到明显差异。
雄性基因敲除小鼠仍具有生育能力，且无任何明显的发育或生精异常，表明在雄性生育方面是可有可无的。
