State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-sen University, Guangzhou 510060, People's Republic of China.
J Genet. 2021;100.
Knockout rats were generated due to its potential involvement in myopia. Subsequent study suggested that the homozygous truncation variants in is associated with male fertility but not myopia. Sperm count and sperm motility were measured in male rats. HE staining, immunofluorescence staining and TUNEL staining were used to observe the gross and histopathology of testis in homozygous and wild rats. knockout rats were generated by CRISPR/Cas9 gene editing. In the process of breeding rats, homozygous male rats were noted to be infertile, with significantly decreased number of sperm suggesting cryptozoospermia that was further confirmed by histologic studies. TUNEL assay showed an increased apoptosis in homozygous testes compared with wild type (<0.001). A significant reduction of spermatocytes was observed in homozygotes compared with wild type (=0.025) by immunofluorescence. These results suggest that is critical for spermatogenesis in male rats.
由于其可能参与近视的发生,敲除大鼠被成功制造。随后的研究表明,基因中的纯合截断变异与男性生育力而非近视有关。对雄性大鼠的精子计数和精子活力进行了测量。使用 HE 染色、免疫荧光染色和 TUNEL 染色观察纯合子和野生型大鼠睾丸的大体和组织病理学变化。使用 CRISPR/Cas9 基因编辑技术成功制造了基因敲除大鼠。在大鼠繁殖过程中,发现纯合雄性大鼠不育,精子数量明显减少,提示存在隐睾症,组织学研究进一步证实了这一点。TUNEL 检测显示,与野生型相比,纯合子睾丸的细胞凋亡增加(<0.001)。通过免疫荧光观察到,与野生型相比,纯合子中的精母细胞数量明显减少(=0.025)。这些结果表明,在雄性大鼠中,对于精子发生,是至关重要的。