基因Cdca2敲除对小鼠精子发生和生育力无明显影响

doi:10.16352/j.issn.1001-6325.2023.05.0717

基础医学与临床 ›› 2023, Vol. 43 ›› Issue (5): 717-723.doi: 10.16352/j.issn.1001-6325.2023.05.0717

基因Cdca2敲除对小鼠精子发生和生育力无明显影响

柳俊, 邹定峰, 缪时英, 宋伟, 李凯^*

中国医学科学院基础医学研究所北京协和医学院基础学院生物化学与分子生物学系医学分子生物学国家重点实验室,北京 100005

收稿日期:2023-01-09 修回日期:2023-02-21 出版日期:2023-05-05 发布日期:2023-04-26
通讯作者: ^*likai@ibms.pumc.edu.cn
基金资助:
国家自然科学基金(31970794,32000586)

Gene Cdca2 knockout has no significant effect on spermatogenesis and fertility in mice

LIU Jun, ZOU Dingfeng, Miao Shiying, SONG Wei, LI Kai^*

State Key Laboratory of Medical Molecular Biology, Department of Biochemistry and Molecular Biology, Institute of Basic Medical Sciences CAMS, School of Basic Medicine PUMC,Beijing 100005, China

Received:2023-01-09 Revised:2023-02-21 Online:2023-05-05 Published:2023-04-26
Contact: ^*likai@ibms.pumc.edu.cn

摘要/Abstract

摘要： 目的探究睾丸高表达细胞分裂周期相关蛋白2基因(Cdca2)在小鼠精子发生和生育力中的生理功能。方法通过Q-PCR和Western blot检测Cdca2在小鼠不同组织部位的表达情况; 利用 CRISPR/Cas9基因编辑技术和Cre-loxP介导的重组系统构建Cdca2组织特异性敲除的小鼠品系;通过苏木精-伊红(HE)染色和形态学分析了解小鼠CDCA2缺失对精子发生的各个阶段生精细胞形态的影响; 通过精子全自动检测分析系统检测小鼠CDCA2缺失对精子活率及运动参数的影响;通过生育力测试实验检测CDCA2缺失对雄性小鼠生育能力的影响。结果 Cdca2是一个睾丸高表达的基因;利用 CRISPR/Cas9技术和生殖系统特异性Cre的工具鼠,成功获得了Cdca2生殖细胞特异性敲除的小鼠;小鼠CDCA2缺失对精子发生中各阶段生精细胞、精子运动参数及雄性生育能力的影响改变无统计学意义。结论基因Cdca2对于小鼠精子发生和雄性生育力的维持可能是非必需的。

Abstract: Objective To explore the physiological function of gene for cell devision cycle associated 2 (Cdca2), a highly expressed testis gene, in spermatogenesis and fertility in mice. Methods The expression of Cdca2 in different tissues of mice was detected by Q-PCR and Western blot; Using CRISPR/Cas9 gene editing technology and Cre-loxP mediated recombination system, a mouse strain with Cdca2 tissue specific knockout was constructed; By hematoxylin-eosin(HE) staining and morphological analysis, the effect of CDCA2 deletion on spermatogenic cell morphology at different stages of spermatogenesis was studied; The effects of CDCA2 deletion on sperm motility and motility parameters were detected by an automatic sperm detection and analysis system; The effect of CDCA2 deletion on the fertility of male mice was examined by fertility test. Results Cdca2 was a highly expressed gene in testis; Cdca2 germ cell specific knockout mice were successfully obtained by CRISPR/Cas9 technology and reproductive system; CDCA2 deletion in mice had no statistically significant effect on spermatogenic cells, sperm motility parameters and male fertility at all stages of spermatogenesis. Conclusions Cdca2 is probably not necessary for spermatogenesis and male fertility maintenance in mice.

Key words: cell division cycle associated 2(CDCA2), RNA binding protein, germ cell, fertility, spermatogenesis

中图分类号:

柳俊, 邹定峰, 缪时英, 宋伟, 李凯. 基因Cdca2敲除对小鼠精子发生和生育力无明显影响[J]. 基础医学与临床, 2023, 43(5): 717-723.

LIU Jun, ZOU Dingfeng, Miao Shiying, SONG Wei, LI Kai. Gene Cdca2 knockout has no significant effect on spermatogenesis and fertility in mice[J]. Basic & Clinical Medicine, 2023, 43(5): 717-723.

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基因Cdca2敲除对小鼠精子发生和生育力无明显影响

Gene Cdca2 knockout has no significant effect on spermatogenesis and fertility in mice

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