1.宜春市妇幼保健院生殖科 江西 宜春 336000
2.武汉大学泰康医学院(基础医学院)/泰康生命医学中心 发育源性疾病湖北省重点实验室 湖北 武汉 430060
段超群,女,1991—,医学硕士,主管技师,主要从事生殖医学相关的研究,E⁃mail:chaoqunduan9@163.com
韩艳,女,1982—,医学硕士,主任医师,主要从事生殖医学相关的研究,E⁃mail:peiyanhan1234@163.com
收稿:2025-03-13,
纸质出版:2025-09-26
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段超群,邓星,孙齐等.Ubap2基因敲除对小鼠精子发生及生育能力的影响[J].武汉大学学报(医学版),2025,46(09):1106-1112.
DUAN Chaoqun,DENG Xing,SUN Qi,et al.Effect of Ubap2 knockout on spermatogenesis and fertility in mice[J].Medical Journal of Wuhan University,2025,46(09):1106-1112.
段超群,邓星,孙齐等.Ubap2基因敲除对小鼠精子发生及生育能力的影响[J].武汉大学学报(医学版),2025,46(09):1106-1112. DOI: 10.14188/j.1671-8852.2025.0216.
DUAN Chaoqun,DENG Xing,SUN Qi,et al.Effect of Ubap2 knockout on spermatogenesis and fertility in mice[J].Medical Journal of Wuhan University,2025,46(09):1106-1112. DOI: 10.14188/j.1671-8852.2025.0216.
目的
2
探究泛素相关蛋白2基因(
Ubap2
)在小鼠精子发生和生育力中的功能。
方法
2
通过RT⁃qPCR和Western Blot分别检测Ubap2 mRNA和蛋白在小鼠不同天数睾丸组织以及不同组织部位的转录和表达情况;利用CRISPR/Cas9基因编辑技术构建
Ubap2
全身性基因敲除(KO)小鼠,通过Sanger测序和Western Blot进行敲除验证;测量
Ubap2
KO小鼠睾丸大小、睾丸⁃体质量比,检测附睾尾精子数量,结合Diff⁃quick染色评估精子形态学特征。通过连续交配实验检测雄性生育指数,检测
Ubap2
缺失对雄性小鼠生育能力的影响;采用减数分裂染色体铺展技术和免疫荧光染色定位、HE染色、TUNEL凋亡检测分析小鼠
Ubap2
KO对精子发生的各个阶段生精细胞发育的影响。
结果
2
Ubap2
在睾丸组织中呈现特异性高表达模式;成功构建了
Ubap2
KO小鼠;
Ubap2
KO小鼠未观察到睾丸发育异常,精子发生关键指标及雄性生育力均未显示统计学差异,生精上皮组织学分析显示各阶段生殖细胞比例与对照组无显著差异。
结论
2
Ubap2
基因对于小鼠精子发生和雄性生育力的维持可能是非必需的。
Objective
2
To investigate the physiological function of ubiquitin⁃associated protein 2 (
Ubap2
) gene in spermatogenesis and fertility in mice.
Methods
2
The expression of Ubap2 in different days and tissues of mice was detected by RT⁃qPCR and Western Blot;
Ubap2
gene knockout (KO) mouse lines were constructed using CRISPR/Cas9 gene editing technology
and the knockout is verified through Sanger sequencing and Western Blot. Testicular size
testis⁃body weight ratio
sperm number and morphology were analyzed
and the effect of
Ubap2
deletion on male mouse fertility was detected through fertility test experiments. The effect of
Ubap2
KO on the development of spermatogenic cells at various stages of spermatogenesis was analyzed through chromosome spreading
immunofluorescence staining
HE staining and TUNEL apoptosis detection.
Results
2
Ubap2
is a gene highly expressed in the testis;
Ubap2
KO mice were successfully constructed using CRISPR/Cas9 technology;
Ubap2
deletion in mice had no statistically significant effect on spermatogenic cells
sperm number
sperm morphology and male fertility at all stages of spermatogenesis.
Conclusion
2
The
Ubap2
gene may be non⁃essential for the maintenance of spermatogenesis and male fertility in mice.
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